KINDAI UNIVERSITY


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TSUBOTA Maho

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FacultyDepartment of Pharmacy
PositionLecturer
Degree
Commentator Guidehttps://www.kindai.ac.jp/meikan/740-tsubota-maho.html
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Last Updated :2020/09/30

Education and Career

Academic & Professional Experience

  •   2018 04 ,  - 現在, Faculty of Pharmacy Department of Pharmacy, Kindai University
  •   2013 04 ,  - 2018 03 , Faculty of Pharmacy Department of Pharmacy, Kindai University
  •   2008 04 ,  - 2013 03 , Faculty of Pharmacy Department of Pharmacy, Kindai University

Research Activities

Research Areas

  • Life sciences, Pharmacology
  • Life sciences, Pharmacology
  • Life sciences, Neuropathology

Research Interests

  • PKA, Cav3.2

Published Papers

  • Genetic deletion of Cav3.2 T-type calcium channels abolishes H2S-dependent somatic and visceral pain signaling in C57BL/6 mice., Kazuki Matsui, Maho Tsubota, Saaya Fukushi, Nene Koike, Hiroshi Masuda, Yoshihito Kasanami, Takaya Miyazaki, Fumiko Sekiguchi, Tsuyako Ohkubo, Shigeru Yoshida, Yutaro Mukai, Akira Oita, Mitsutaka Takada, Atsufumi Kawabata, Journal of pharmacological sciences, Journal of pharmacological sciences, 140(3), 310 - 312, Jul. 2019 , Refereed
    Summary:We tested whether genetic deletion of Cav3.2 T-type Ca2+ channels abolishes hydrogen sulfide (H2S)-mediated pain signals in mice. In Cav3.2-expressing HEK293 cells, Na2S, an H2S donor, at 100 μM clearly increased Ba2+ currents, as assessed by whole-cell patch-clamp recordings. In wild-type C57BL/6 mice, intraplantar and intracolonic administration of Na2S evoked mechanical allodynia and visceral nociceptive behavior, respectively, which were abolished by TTA-A2, a T-type Ca2+ channel blocker. In Cav3.2-knockout mice of a C57BL/6 background, Na2S caused neither somatic allodynia nor colonic nociception. Our study thus provides definitive evidence for an essential role of Cav3.2 in H2S-dependent somatic and colonic pain.
  • Involvement of the cystathionine-γ-lyase/Cav3.2 pathway in substance P-induced bladder pain in the mouse, a model for nonulcerative bladder pain syndrome., Maho Tsubota, Yasumasa Okawa, Yuhei Irie, Mariko Maeda, Tomoka Ozaki, Fumiko Sekiguchi, Hiroyasu Ishikura, Atsufumi Kawabata, Neuropharmacology, Neuropharmacology, 133, 254 - 263, May 01 2018 , Refereed
    Summary:Hydrogen sulfide (H2S) formed by cystathionine-γ-lyase (CSE) enhances the activity of Cav3.2 T-type Ca2+ channels, contributing to the bladder pain accompanying hemorrhagic cystitis caused by systemic administration of cyclophosphamide (CPA) in mice. Given clinical and fundamental evidence for the involvement of the substance P/NK1 receptor systems in bladder pain syndrome (BPS)/interstitial cystitis (IC), we created an intravesical substance P-induced bladder pain model in mice and analyzed the possible involvement of the CSE/Cav3.2 pathway. Bladder pain/cystitis was induced by i.p. CPA or intravesical substance P in female mice. Bladder pain was evaluated by counting nociceptive behavior and by detecting referred hyperalgesia in the lower abdomen and hindpaw. The isolated bladder tissue was weighed to estimate bladder swelling and subjected to histological observation and Western blotting. Intravesical substance P caused profound referred hyperalgesia accompanied by little bladder swelling or edema 6-24 h after the administration, in contrast to i.p. CPA-induced nociceptive behavior/referred hyperalgesia with remarkable bladder swelling/edema and urothelial damage. The bladder pain and/or cystitis symptoms caused by substance P or CPA were prevented by the NK1 receptor antagonist. CSE in the bladder was upregulated by substance P or CPA, and the NK1 antagonist prevented the CPA-induced CSE upregulation. A CSE inhibitor, a T-type Ca2+ channel blocker and gene silencing of Cav3.2 abolished the intravesical substance P-induced referred hyperalgesia. The intravesical substance P-induced pain in mice is useful as a model for nonulcerative BPS, and involves the activation of the NK1 receptor/CSE/H2S/Cav3.2 cascade.
  • Involvement of NF-κB in the upregulation of cystathionine-γ-lyase, a hydrogen sulfide-forming enzyme, and bladder pain accompanying cystitis in mice., Tomoka Ozaki, Maho Tsubota, Fumiko Sekiguchi, Atsufumi Kawabata, Clinical and experimental pharmacology & physiology, Clinical and experimental pharmacology & physiology, 45(4), 355 - 361, Apr. 2018 , Refereed
    Summary:Hydrogen sulfide (H2 S) is generated from l-cysteine by multiple enzymes including cystathionine-γ-lyase (CSE), and promotes nociception by targeting multiple molecules such as Cav 3.2 T-type Ca2+ channels. Bladder pain accompanying cyclophosphamide (CPA)-induced cystitis in mice has been shown to involve the functional upregulation of the CSE/H2 S/Cav 3.2 pathway. Therefore, we investigated whether NF-κB, as an upstream signal of the CSE/H2 S system, contributes to bladder pain in mice with CPA-induced cystitis. Bladder pain-like nociceptive behaviour was observed in CPA-treated mice, and referred hyperalgesia was evaluated by the von Frey test. Isolated bladder weights were assessed to estimate bladder swelling, and protein levels were measured by Western blotting. CPA, administered intraperitoneally, induced nociceptive behaviour, referred hyperalgesia and increased bladder weights in mice. β-Cyano-l-alanine, a reversible selective CSE inhibitor, prevented CPA-induced nociceptive behaviour, referred hyperalgesia, and, in part, increases in bladder weight. CPA markedly increased phosphorylated NF-κB p65 levels in the bladder, an effect that was prevented by pyrrolidine dithiocarbamate (PDTC), an NF-κB inhibitor. PDTC and curcumin, which inhibits NF-κB signals, abolished CPA-induced nociceptive behaviour, referred hyperalgesia and, in part, increases in bladder weight. CPA caused the overexpression of CSE in the bladder, and this was prevented by PDTC or curcumin. The CPA-induced activation of NF-κB signals appeared to cause CSE overexpression in the bladder, contributing to bladder pain and in part swelling, possibly through H2 S/Cav 3.2 signaling. Therefore, NF-κB-inhibiting compounds including curcumin may be useful for the treatment of cystitis-related bladder pain.
  • Involvement of macrophage-derived HMGB1 in acute pancreatits-related pain: roles of RAGE and cnR4 as the target molecules, Irie Yuhei, Tsubota Maho, Sekiguchi Fumiko, Ishikura Hiroyasu, Nishibori Masahiro, Kawabata Atsufumi, JOURNAL OF PHARMACOLOGICAL SCIENCES, JOURNAL OF PHARMACOLOGICAL SCIENCES, 130(3), S81, Mar. 2016 , Refereed
  • Zinc deficiency-induced hyperalgesia in mice: involvement of Ca(v)3.2 and high mobility group box 1, Tomita Shiori, Shikimi Shiyu, Sekiguchi Fumiko, Tsubota Maho, Shirai Akihiro, Nishibori Masahiro, Kawabata Atsufumi, JOURNAL OF PHARMACOLOGICAL SCIENCES, JOURNAL OF PHARMACOLOGICAL SCIENCES, 130(3), S82, Mar. 2016 , Refereed
  • Zinc chelator aggravates bladder pain accompanying cyclophosphamide-induced cystitis by enhancing T-type Ca2+ channel activity, Matsuoka Junki, Ozaki Tomoka, Tsubota Maho, Kawabata Atsufumi, JOURNAL OF PHARMACOLOGICAL SCIENCES, JOURNAL OF PHARMACOLOGICAL SCIENCES, 130(3), S200, Mar. 2016 , Refereed
  • The hyperalgesia induced by peripheral HMGB1 in distinct redox states involves different mechanisms, Tsubota Maho, Yamasoba Daichi, Domoto Risa, Sekiguchi Fumiko, Nishibori Masahiro, Kawabata Atsufumi, JOURNAL OF PHARMACOLOGICAL SCIENCES, JOURNAL OF PHARMACOLOGICAL SCIENCES, 130(3), S235, Mar. 2016 , Refereed
  • Roles of macrophage-derived HMGB1 in a substance P-induced interstitial cystitis/bladder pain syndrome mouse model, Maeda Mariko, Irie Yuhei, Tsubota Maho, Kubo Lisa, Sekiguchi Fumiko, Ishikura Hiroyasu, Nishibori Masahiro, Kawabata Atsufumi, JOURNAL OF PHARMACOLOGICAL SCIENCES, JOURNAL OF PHARMACOLOGICAL SCIENCES, 130(3), S144, Mar. 2016 , Refereed
  • Anti-HMGB1 therapy for visceral pain, Tsubota Maho, Kawabata Atsufumi, JOURNAL OF PHARMACOLOGICAL SCIENCES, JOURNAL OF PHARMACOLOGICAL SCIENCES, 128(3), S40, Jul. 2015 , Refereed
  • Preventive and therapeutic effects of polaprezinc, a zinc-containing drug, on cyclophosphamide-induced cystitis and/or bladder pain, Hiruma Saki, Murakami, Nakayama) Masahiro, Tsubota Maho, Sekiguchi Fumiko, Matsuyama Kenji, Kimura Takeshi, Moriyama Masahiro, Kawabata Atsufumi, JOURNAL OF PHARMACOLOGICAL SCIENCES, JOURNAL OF PHARMACOLOGICAL SCIENCES, 128(3), S110, Jul. 2015 , Refereed
  • Macrophage-derived HMGB1 contributes to the inflammatory hyperalgegia through the NF-kappa B pathway, Yamasoba Daichi, Seki Yukari, Yamanishi Hiroki, Tsubota Maho, Sekiguchi Fumiko, Yagi Hideki, Masuko Takashi, Nishibori Masahiro, Kawabata Atsufumi, JOURNAL OF PHARMACOLOGICAL SCIENCES, JOURNAL OF PHARMACOLOGICAL SCIENCES, 128(3), S104, Jul. 2015 , Refereed
  • [Proteinase-activated receptor]., Matsunami M, Sekiguchi T, Kawabata A, Nihon yakurigaku zasshi. Folia pharmacologica Japonica, Nihon yakurigaku zasshi. Folia pharmacologica Japonica, 128(6), 434 - 436, Dec. 2006 , Refereed
  • Cystitis-Related Bladder Pain Involves ATP-Dependent HMGB1 Release from Macrophages and Its Downstream H2S/Cav3.2 Signaling in Mice., Shiori Hiramoto, Maho Tsubota, Kaoru Yamaguchi, Kyoko Okazaki, Aya Sakaegi, Yuki Toriyama, Junichi Tanaka, Fumiko Sekiguchi, Hiroyasu Ishikura, Hidenori Wake, Masahiro Nishibori, Huy Du Nguyen, Takuya Okada, Naoki Toyooka, Atsufumi Kawabata, Cells, Cells, 9(8), Jul. 22 2020 , Refereed
    Summary:Cystitis-related bladder pain involves RAGE activation by HMGB1, and increased Cav3.2 T-type Ca2+ channel activity by H2S, generated by upregulated cystathionine-γ-lyase (CSE) in mice treated with cyclophosphamide (CPA). We, thus, investigated possible crosstalk between the HMGB1/RAGE and CSE/H2S/Cav3.2 pathways in the bladder pain development. Bladder pain (nociceptive behavior/referred hyperalgesia) and immuno-reactive CSE expression in the bladder were determined in CPA-treated female mice. Cell signaling was analyzed in urothelial T24 and macrophage-like RAW264.7 cells. The CPA-induced bladder pain was abolished by pharmacological inhibition of T-type Ca2+ channels or CSE, and genetic deletion of Cav3.2. The CPA-induced CSE upregulation, as well as bladder pain was prevented by HMGB1 inactivation, inhibition of HMGB1 release from macrophages, antagonists of RAGE or P2X4/P2X7 receptors, and N-acetylcysteine, an antioxidant. Acrolein, a metabolite of CPA, triggered ATP release from T24 cells. Adenosine triphosphate (ATP) stimulated cell migration via P2X7/P2X4, and caused HMGB1 release via P2X7 in RAW264.7 cells, which was dependent on p38MAPK/NF-κB signaling and reactive oxygen species (ROS) accumulation. Together, our data suggest that CPA, once metabolized to acrolein, causes urothelial ATP-mediated, redox-dependent HMGB1 release from macrophages, which in turn causes RAGE-mediated CSE upregulation and subsequent H2S-targeted Cav3.2-dependent nociceptor excitation, resulting in bladder pain.
  • HMGB1 and its membrane receptors as therapeutic targets in an intravesical substance P-induced bladder pain syndrome mouse model., Yuhei Irie, Maho Tsubota, Mariko Maeda, Shiori Hiramoto, Fumiko Sekiguchi, Hiroyasu Ishikura, Hidenori Wake, Masahiro Nishibori, Atsufumi Kawabata, Journal of pharmacological sciences, Journal of pharmacological sciences, 143(2), 112 - 116, Jun. 2020 , Refereed
    Summary:HMGB1, a nuclear protein, once released to the extracellular space, promotes somatic and visceral pain signals. We thus analyzed the role of HMGB1 in an intravesical substance P-induced bladder pain syndrome (BPS) mouse model. Intravesical administration of substance P caused referred hyperalgesia/allodynia in the lower abdomen and hindpaw without producing severe urothelial damage, which was prevented by an anti-HMGB1-neutralizing antibody, thrombomodulin α capable of inactivating HMGB1 and antagonists of RAGE or CXCR4. The HMGB1 inactivation or RAGE blockade also reversed the established bladder pain symptoms. HMGB1 and RAGE are thus considered to serve as therapeutic targets for BPS.
  • Role of high-mobility group box 1 and its modulation by thrombomodulin/thrombin axis in neuropathic and inflammatory pain., Ryuichi Tsujita, Maho Tsubota, Fumiko Sekiguchi, Atsufumi Kawabata, British journal of pharmacology, British journal of pharmacology, May 06 2020 , Refereed
    Summary:High-mobility group box 1 (HMGB1), a nuclear protein, once released to the extracellular space, facilitates pain signals as well as inflammation. Intraplantar or intraspinal application of HMGB1 elicits hyperalgesia/allodynia in rodents by activating the advanced glycosylation end-product specific receptor (receptor for advanced glycation end-products; RAGE) or Toll-like receptor 4 (TLR4). Endogenous HMGB1 derived from neurons, perineuronal cells or immune cells accumulating in the dorsal root ganglion or sensory nerves participates in somatic and visceral pain consisting of neuropathic and/or inflammatory components. Endothelial thrombomodulin (TM) and recombinant human soluble TM, TMα, markedly increase thrombin-dependent degradation of HMGB1, and systemic administration of TMα prevents and reverses various HMGB1-dependent pathological pain. Low MW compounds that directly inactivate HMGB1 or antagonize HMGB1-targeted receptors would be useful to reduce various forms of intractable pain. Thus, HMGB1 and its receptors are considered to serve as promising targets in developing novel agents to prevent or treat pathological pain.
  • Tacrolimus, a calcineurin inhibitor, promotes capsaicin-induced colonic pain in mice., Kazuki Matsui, Yuka Terada, Maho Tsubota, Fumiko Sekiguchi, Atsufumi Kawabata, Journal of pharmacological sciences, Journal of pharmacological sciences, 143(1), 60 - 63, May 2020 , Refereed
    Summary:TRPV1 is phosphorylated and functionally upregulated by protein kinases, and negatively regulated by phosphatases including calcineurin. Since the clinical use of calcineurin-inhibiting immunosuppressants is commonly associated with chronic diarrhea, we examined if tacrolimus, a calcineurin inhibitor, promotes TRPV1-dependent colonic hypersensitivity in mice. Intracolonic administration of capsaicin, a TRPV1 agonist, caused referred hyperalgesia in the lower abdomen, an effect prevented by capsazepine, a TRPV1 blocker. Tacrolimus accelerated the intracolonic capsaicin-induced referred hyperalgesia. Similarly, intracolonic capsaicin caused spinal ERK phosphorylation, a marker for nociceptor excitation, an effect promoted by tacrolimus. Thus, tacrolimus may aggravate TRPV1-related colonic pain accompanying irritable bowel syndrome.
  • Role of non-macrophage cell-derived HMGB1 in oxaliplatin-induced peripheral neuropathy and its prevention by the thrombin/thrombomodulin system in rodents: negative impact of anticoagulants., Maho Tsubota, Ryotaro Fukuda, Yusuke Hayashi, Takaya Miyazaki, Shin Ueda, Rika Yamashita, Nene Koike, Fumiko Sekiguchi, Hidenori Wake, Shuji Wakatsuki, Yuka Ujiie, Toshiyuki Araki, Masahiro Nishibori, Atsufumi Kawabata, Journal of neuroinflammation, Journal of neuroinflammation, 16(1), 199 - 199, Oct. 30 2019 , Refereed
    Summary:BACKGROUND: Macrophage-derived high mobility group box 1 (HMGB1), a damage-associated molecular pattern (DAMP) protein, plays a key role in the development of chemotherapy-induced peripheral neuropathy (CIPN) caused by paclitaxel in rodents. Endothelial thrombomodulin (TM) promotes thrombin-induced degradation of HMGB1, and TMα, a recombinant human soluble TM, abolishes peripheral HMGB1-induced allodynia in mice. We thus examined whether HMGB1, particularly derived from macrophages, contributes to oxaliplatin-induced neuropathy in mice and analyzed the anti-neuropathic activity of the TM/thrombin system. METHODS: CIPN models were created by the administration of oxaliplatin in mice and rats, and the nociceptive threshold was assessed by von Frey test or paw pressure test. Macrophage-like RAW264.7 cells were stimulated with oxaliplatin in vitro. Proteins were detected and/or quantified by Western blotting, immunostaining, or enzyme-linked immunosorbent assay. RESULTS: Intraperitoneal administration of an anti-HMGB1-neutralizing antibody (AB) at 1 mg/kg prevented the oxaliplatin-induced allodynia in mice and rats. Antagonists of Toll-like receptor (TLR) 4, receptor for advanced glycation end products (RAGE) and CXCR4 among the HMGB1-targeted pro-nociceptive receptors, also mimicked the anti-neuropathic activity of AB in mice. Macrophage accumulation in the sciatic nerve was observed in mice treated with paclitaxel, but not oxaliplatin, and neither macrophage depletion nor inhibitors of macrophage activation affected oxaliplatin-induced allodynia. Oxaliplatin was 10- to 100-fold less potent than paclitaxel in releasing HMGB1 from macrophage-like RAW264.7 cells. Like AB, TMα at 10 mg/kg prevented the oxaliplatin-induced allodynia in mice as well as rats, an effect abolished by argatroban at 10 mg/kg, a thrombin inhibitor. The anti-neuropathic activity of TMα in oxaliplatin-treated mice was suppressed by oral anticoagulants such as warfarin at 1 mg/kg, dabigatran at 75 mg/kg, and rivaroxaban at 10 mg/kg, but not antiplatelet agents such as aspirin at 50 mg/kg and clopidogrel at 10 mg/kg. Repeated administration of the anticoagulants gradually developed neuropathic allodynia and elevated plasma HMGB1 levels in mice treated with a subeffective dose of oxaliplatin. CONCLUSIONS: Our data thus suggests a causative role of HMGB1 derived from non-macrophage cells in oxaliplatin-induced peripheral neuropathy and a thrombin-dependent anti-neuropathic activity of exogenous TMα and, most probably, endogenous TM.
  • NNC 55-0396, a T-type calcium channel blocker, protects against the brain injury induced by middle cerebral artery occlusion and reperfusion in mice., Sachi Matsuda, Hiroyuki Nishikawa, Anna Fukatsu, Yuko Kurokawa, Maho Tsubota, Fumiko Sekiguchi, Shogo Tokuyama, Atsufumi Kawabata, Journal of pharmacological sciences, Journal of pharmacological sciences, 140(2), 193 - 196, Jun. 2019 , Refereed
    Summary:We tested whether NNC 55-0396 (NNC), a T-type calcium channel (T-channel) blocker, reduces the brain injury caused by middle cerebral artery occlusion and reperfusion (MCAO/R) in mice. NNC, administered i.c.v. before the occlusion, greatly reduced the MCAO/R-induced brain infarct and neurological dysfunctions, although it, given toward the end of occlusion, was less effective. Systemic administration of NNC before the occlusion also attenuated the infarct and neurological dysfunctions. Our data imply that blood-brain-barrier-permeable T-channel blockers such as NNC are capable of reducing MCAO/R-induced brain damage, and that T-channels are involved in neuronal damage induced by ischemia rather than reperfusion.
  • Dietary ascorbic acid restriction in GNL/SMP30-knockout mice unveils the role of ascorbic acid in regulation of somatic and visceral pain sensitivity., Maho Tsubota, Kenta Uebo, Koki Miki, Fumiko Sekiguchi, Akihiko Ishigami, Atsufumi Kawabata, Biochemical and biophysical research communications, Biochemical and biophysical research communications, 511(3), 705 - 710, Apr. 09 2019 , Refereed
    Summary:Cav3.2 T-type Ca2+ channels are expressed in the primary afferents and play a pronociceptive role. The activity of Cav3.2 is enhanced by H2S, a gasotransmitter, and suppressed by ascorbic acid (vitamin C) through metal-catalyzed oxidation of the Zn2+-binding His191 in Cav3.2. Since rodents, but not humans, are capable of synthesizing ascorbic acid, the present study examined the role of ascorbic acid in nociceptive processing, using the mice lacking GNL/SMP30, an enzyme essential for ascorbic acid biosynthesis. Intraplantar and intracolonic administration of NaHS, an H2S donor, caused somatic allodynia and referred hyperalgesia, respectively, and repeated treatment with paclitaxel produced neuropathic allodynia in wild-type mice, all of which were suppressed by ascorbic acid or T-type Ca2+ channel blockers. Dietary ascorbic acid restriction caused dramatic decreases in plasma and tissue ascorbic acid levels in GNL/SMP30-knockout, but not wild-type, mice. The ascorbic acid restriction enhanced the somatic and visceral hypersensitivity following intraplantar and intracolonic NaHS, respectively, and paclitaxel-induced neuropathy in GNL/SMP30-knockout mice, while it had no such effect in wild-type mice. Together, our data unveil the critical role of ascorbic acid in regulating somatic and visceral pain sensitivity and support accumulating clinical evidence for the usefulness of ascorbic acid in pain management.
  • Critical role of Cav3.2 T-type calcium channels in the peripheral neuropathy induced by bortezomib, a proteasome-inhibiting chemotherapeutic agent, in mice., Shiori Tomita, Fumiko Sekiguchi, Tomoyo Deguchi, Takaya Miyazaki, Yuya Ikeda, Maho Tsubota, Shigeru Yoshida, Huy Du Nguyen, Takuya Okada, Naoki Toyooka, Atsufumi Kawabata, Toxicology, Toxicology, 413, 33 - 39, Feb. 01 2019 , Refereed
    Summary:Bortezomib, a first-line agent for treatment of multiple myeloma, exhibits anticancer activity through proteasome inhibition. However, bortezomib-induced peripheral neuropathy (BIPN) is one of the most serious side effects. Since decreased proteasomal degradation of Cav3.2 T-type calcium channels in the primary afferents is involved in persistent pain, we investigated whether BIPN involves increased protein levels of Cav3.2 in mice. Six repeated i.p. administrations of bortezomib for 12 days developed persistent mechanical allodynia. Systemic administration of novel T-type calcium channel blockers, (2R/S)-6-prenylnaringenin and KTt-45, and of TTA-A2, the well-known blocker, reversed the BIPN. Ascorbic acid, known to block Cav3.2, but not Cav3.1 or 3.3, and silencing of Cav3.2 gene also suppressed BIPN. Protein levels of Cav3.2 in the dorsal root ganglion (DRG) at L4-L6 levels increased throughout days 1-21 after the onset of bortezomib treatment. Protein levels of USP5, a deubiquitinating enzyme that specifically inhibits proteasomal degradation of Cav3.2, increased in DRG on days 3-21, but not day 1, in bortezomib-treated mice. In DRG-derived ND7/23 cells, bortezomib increased protein levels of Cav3.2 and T-channel-dependent currents, as assessed by a patch-clamp method, but did not upregulate expression of Cav3.2 mRNA or USP5 protein. MG-132, another proteasome inhibitor, also increased Cav3.2 protein levels in the cultured cells. Given the previous evidence for USP5 induction following nociceptor excitation, our data suggest that BIPN involves the increased protein levels of Cav3.2 in nociceptors through inhibition of proteasomal degradation of Cav3.2 by bortezomib itself and then by USP5 that is upregulated probably in an activity-dependent manner.
  • [Regulation of Cav3.2-mediated pain signals by hydrogen sulfide]., Maho Tsubota, Atsufumi Kawabata, Nihon yakurigaku zasshi. Folia pharmacologica Japonica, Nihon yakurigaku zasshi. Folia pharmacologica Japonica, 154(3), 128 - 132, 2019 , Refereed
    Summary:Hydrogen sulfide (H2S), an endogenous gasotransmitter, is generated from L-cysteine by 3 distinct enzymes including cystathionine-γ-lyase (CSE), and targets multiple molecules, thereby playing various roles in health and disease. H2S triggers or accelerates somatic pain and visceral nociceptive signals in the pancreas, colon and bladder by enhancing the activity of Cav3.2 T-type calcium channels. H2S also activates TRPA1, which participates in H2S-induced somatic pain signaling. However, Cav3.2 predominantly mediates colonic nociception by H2S, because genetic deletion of TRPA1 does not reduce H2S-induced colonic pain. The functional upregulation of the CSE/H2S/Cav3.2 system is involved in neuropathic pain and visceral pain accompanying pancreatitis and cystitis. Cav3.2 also appears to participate in irritable bowel syndrome (IBS), although the role of endogenous H2S generation by CSE in IBS is still open to question. In this review, we describe how H2S regulates pain signals, particularly by interacting with Cav3.2.
  • [HMGB1 as a target for prevention of chemotherapy-induced peripheral neuropathy]., Atsufumi Kawabata, Maho Tsubota, Fumiko Sekiguchi, Ryuichi Tsujita, Nihon yakurigaku zasshi. Folia pharmacologica Japonica, Nihon yakurigaku zasshi. Folia pharmacologica Japonica, 154(5), 236 - 240, 2019 , Refereed
    Summary:Chemotherapy-induced peripheral neuropathy (CIPN) considerably impairs cancer patients' QOL, and may lead to discontinuation of drug treatment of cancer. Currently, there is no effective strategy against CIPN. Therefore, it is an urgent issue to develop clinically available drugs that prevent or treat CIPN. We have shown that high mobility group box 1 (HMGB1), a damage-associated molecular pattern (DAMP) molecule, plays an essential role in the development of CIPN. Most interestingly, thrombomodulin α, approved as a medicine for treatment of disseminated intravascular coagulation (DIC) in Japan, causes thrombin-dependent degradation of extracellular HMGB1 that is released in response to chemotherapeutics, and prevents CIPN. Thus, we expect that targeting HMGB1 or its receptors would lead to prevention of CIPN in cancer patients in near future.
  • Paclitaxel-induced HMGB1 release from macrophages and its implication for peripheral neuropathy in mice: Evidence for a neuroimmune crosstalk., Fumiko Sekiguchi, Risa Domoto, Kana Nakashima, Daichi Yamasoba, Hiroki Yamanishi, Maho Tsubota, Hidenori Wake, Masahiro Nishibori, Atsufumi Kawabata, Neuropharmacology, Neuropharmacology, 141, 201 - 213, Oct. 2018 , Refereed
    Summary:Given our recent evidence for the role of high mobility group box 1 (HMGB1) in chemotherapy-induced peripheral neuropathy (CIPN) in rats, we examined the origin of HMGB1 and the upstream and downstream mechanisms of HMGB1 release involved in paclitaxel-induced neuropathy in mice. Paclitaxel treatment developed mechanical allodynia in mice, as assessed by von Frey test, which was prevented by an anti-HMGB1-neutralizing antibody or thrombomodulin alfa capable of inactivating HMGB1. RAGE or CXCR4 antagonists, ethyl pyruvate or minocycline, known to inhibit HMGB1 release from macrophages, and liposomal clodronate, a macrophage depletor, prevented the paclitaxel-induced allodynia. Paclitaxel caused upregulation of RAGE and CXCR4 in the dorsal root ganglia and macrophage accumulation in the sciatic nerve. In macrophage-like RAW264.7 cells, paclitaxel evoked cytoplasmic translocation of nuclear HMGB1 followed by its extracellular release, and overexpression of CBP and PCAF, histone acetyltransferases (HATs), known to cause acetylation and cytoplasmic translocation of HMGB1, which were suppressed by ethyl pyruvate, N-acetyl-l-cysteine, an anti-oxidant, and SB203580 and PDTC, inhibitors of p38 MAP kinase (p38MAPK) and NF-κB, respectively. Paclitaxel increased accumulation of reactive oxygen species (ROS) and phosphorylation of p38MAPK, NF-κB p65 and I-κB in RAW264.7 cells. In mice, N-acetyl-l-cysteine or PDTC prevented the paclitaxel-induced allodynia. Co-culture of neuron-like NG108-15 cells or stimulation with their conditioned medium promoted paclitaxel-induced HMGB1 release from RAW264.7 cells. Our data indicate that HMGB1 released from macrophages through the ROS/p38MAPK/NF-κB/HAT pathway participates in the paclitaxel-induced peripheral neuropathy in mice, and unveils an emerging therapeutic avenue targeting a neuroimmune crosstalk in CIPN.
  • Design and synthesis of novel anti-hyperalgesic agents based on 6-prenylnaringenin as the T-type calcium channel blockers., Huy Du Nguyen, Takuya Okada, Shun Kitamura, Sakura Yamaoka, Yamato Horaguchi, Yoshihito Kasanami, Fumiko Sekiguchi, Maho Tsubota, Shigeru Yoshida, Hiroyuki Nishikawa, Atsufumi Kawabata, Naoki Toyooka, Bioorganic & medicinal chemistry, Bioorganic & medicinal chemistry, 26(15), 4410 - 4427, Aug. 15 2018 , Refereed
    Summary:Since 6-prenylnaringenin (6-PNG) was recently identified as a novel T-type calcium channel blocker with the IC50 value around 1 µM, a series of flavanone derivatives were designed, synthesized and subsequently evaluated for T-channel-blocking activity in HEK293 cells transfected with Cav3.2 T-type channels using a patch-clamp technique. As a result, several new flavanones blocked Cav3.2-dependent T-currents more potently than 6-PNG. In the synthesized compounds, 6-(3-ethylpent-2-enyl)-5,7-dihydroxy-2-(2-hydroxyphenyl)chroman-4-one 8j, 6-(3-ethylpent-2-enyl)-5,7-dihydroxy-2-(4-hydroxyphenyl)chroman-4-one 11b, 6-(2-cyclopentylideneethyl)-5,7-dihydroxy-2-(4-hydroxyphenyl)chroman-4-one 11d, and 6-(2-Cyclopentylethyl)-5,7-dihydroxy-2-(4-hydroxyphenyl)chroman-4-one 12c were more potent blocker than 6-PNG with the IC50 value of 0.39, 0.26, 0.46, and 0.50 µM, respectively. Among the above four derivatives, the compound 8j provided the best result in the in vivo experiments; i.e. systemic administration of 8j at the minimum dose completely restored neuropathic pain induced by partial sciatic nerve ligation in mice.
  • Blockade of T-type calcium channels by 6-prenylnaringenin, a hop component, alleviates neuropathic and visceral pain in mice., Fumiko Sekiguchi, Tomoyo Fujita, Takahiro Deguchi, Sakura Yamaoka, Ken Tomochika, Maho Tsubota, Sumire Ono, Yamato Horaguchi, Maki Ichii, Mio Ichikawa, Yumiko Ueno, Nene Koike, Tadatoshi Tanino, Huy Du Nguyen, Takuya Okada, Hiroyuki Nishikawa, Shigeru Yoshida, Tsuyako Ohkubo, Naoki Toyooka, Kazuya Murata, Hideaki Matsuda, Atsufumi Kawabata, Neuropharmacology, Neuropharmacology, 138, 232 - 244, Aug. 2018 , Refereed
    Summary:Since Cav3.2 T-type Ca2+ channels (T-channels) expressed in the primary afferents and CNS contribute to intractable pain, we explored T-channel-blocking components in distinct herbal extracts using a whole-cell patch-clamp technique in HEK293 cells stably expressing Cav3.2 or Cav3.1, and purified and identified sophoraflavanone G (SG) as an active compound from SOPHORAE RADIX (SR). Interestingly, hop-derived SG analogues, (2S)-6-prenylnaringenin (6-PNG) and (2S)-8-PNG, but not naringenin, also blocked T-channels; IC50 (μM) of SG, (2S)-6-PNG and (2S)-8-PNG was 0.68-0.75 for Cav3.2 and 0.99-1.41 for Cav3.1. (2S)-6-PNG and (2S)-8-PNG, but not SG, exhibited reversible inhibition. The racemic (2R/S)-6-PNG as well as (2S)-6-PNG potently blocked Cav3.2, but exhibited minor effect on high-voltage-activated Ca2+ channels and voltage-gated Na+ channels in differentiated NG108-15 cells. In mice, the mechanical allodynia following intraplantar (i.pl.) administration of an H2S donor was abolished by oral or i.p. SR extract and by i.pl. SG, (2S)-6-PNG or (2S)-8-PNG, but not naringenin. Intraperitoneal (2R/S)-6-PNG strongly suppressed visceral pain and spinal ERK phosphorylation following intracolonic administration of an H2S donor in mice. (2R/S)-6-PNG, administered i.pl. or i.p., suppressed the neuropathic allodynia induced by partial sciatic nerve ligation or oxaliplatin, an anti-cancer agent, in mice. (2R/S)-6-PNG had little or no effect on open-field behavior, motor performance or cardiovascular function in mice, and on the contractility of isolated rat aorta. (2R/S)-6-PNG, but not SG, was detectable in the brain after their i.p. administration in mice. Our data suggest that 6-PNG, a hop component, blocks T-channels, and alleviates neuropathic and visceral pain with little side effects.
  • Role of Thrombin in Soluble Thrombomodulin-Induced Suppression of Peripheral HMGB1-Mediated Allodynia in Mice., Ryuichi Tsujita, Maho Tsubota, Yusuke Hayashi, Haruka Saeki, Fumiko Sekiguchi, Atsufumi Kawabata, Journal of neuroimmune pharmacology : the official journal of the Society on NeuroImmune Pharmacology, Journal of neuroimmune pharmacology : the official journal of the Society on NeuroImmune Pharmacology, 13(2), 179 - 188, Jun. 2018 , Refereed
    Summary:High mobility group box 1 (HMGB1), a nuclear protein, once released into the extracellular space under pathological conditions, plays a pronociceptive role in redox-dependent distinct active forms, all-thiol HMGB1 (at-HMGB1) and disulfide HMGB1 (ds-HMGB1), that accelerate nociception through the receptor for advanced glycation endproducts (RAGE) and Toll-like receptor 4 (TLR4), respectively. Thrombomodulin (TM), an endothelial membrane protein, and soluble TM, known as TMα, promote thrombin-mediated activation of protein C and also sequester HMGB1, which might facilitate thrombin degradation of HMGB1. The present study aimed at clarifying the role of thrombin in TMα-induced suppression of peripheral HMGB1-dependent allodynia in mice. Thrombin-induced degradation of at-HMGB1 and ds-HMGB1 was accelerated by TMα in vitro. Intraplantar (i.pl.) injection of bovine thymus-derived HMGB1 in an unknown redox state, at-HMGB1, ds-HMGB1 or lipopolysaccharide (LPS), known to cause HMGB1 secretion, produced long-lasting mechanical allodynia in mice, as assessed by von Frey test. TMα, when preadministered i.pl., prevented the allodynia caused by bovine thymus-derived HMGB1, at-HMGB1, ds-HMGB1 or LPS, in a dose-dependent manner. The TMα-induced suppression of the allodynia following i.pl. at-HMGB1, ds-HMGB1 or LPS was abolished by systemic preadministration of argatroban, a thrombin-inhibiting agent, and accelerated by i.pl. co-administered thrombin. Our data clearly indicate that TMα is capable of promoting the thrombin-induced degradation of both at-HMGB1 and ds-HMGB1, and suppresses the allodynia caused by either HMGB1 in a thrombin-dependent manner. Considering the emerging role of HMGB1 in distinct pathological pain models, the present study suggests the therapeutic usefulness of TMα for treatment of intractable and/or persistent pain.
  • Zinc deficiency promotes cystitis-related bladder pain by enhancing function and expression of Cav3.2 in mice., Tomoka Ozaki, Junki Matsuoka, Maho Tsubota, Shiori Tomita, Fumiko Sekiguchi, Takeshi Minami, Atsufumi Kawabata, Toxicology, Toxicology, 393, 102 - 112, Jan. 15 2018 , Refereed
    Summary:Cav3.2 T-type Ca2+ channel activity is suppressed by zinc that binds to the extracellular histidine-191 of Cav3.2, and enhanced by H2S that interacts with zinc. Cav3.2 in nociceptors is upregulated in an activity-dependent manner. The enhanced Cav3.2 activity by H2S formed by the upregulated cystathionine-γ-lyase (CSE) is involved in the cyclophosphamide (CPA)-induced cystitis-related bladder pain in mice. We thus asked if zinc deficiency affects the cystitis-related bladder pain in mice by altering Cav3.2 function and/or expression. Dietary zinc deficiency for 2 weeks greatly decreased zinc concentrations in the plasma but not bladder tissue, and enhanced the bladder pain/referred hyperalgesia (BP/RH) following CPA at 200mg/kg, a subeffective dose, but not 400mg/kg, a maximal dose, an effect abolished by pharmacological blockade or gene silencing of Cav3.2. Acute zinc deficiency caused by systemic N,N,N',N'-tetrakis-(2-pyridylmethyl)-ethylendiamine (TPEN), a zinc chelator, mimicked the dietary zinc deficiency-induced Cav3.2-dependent promotion of BP/RH following CPA at 200mg/kg. CPA at 400mg/kg alone or TPEN plus CPA at 200mg/kg caused Cav3.2 overexpression accompanied by upregulation of Egr-1 and USP5, known to promote transcriptional expression and reduce proteasomal degradation of Cav3.2, respectively, in the dorsal root ganglia (DRG). The CSE inhibitor, β-cyano-l-alanine, prevented the BP/RH and upregulation of Cav3.2, Egr-1 and USP5 in DRG following TPEN plus CPA at 200mg/kg. Together, zinc deficiency promotes bladder pain accompanying CPA-induced cystitis by enhancing function and expression of Cav3.2 in nociceptors, suggesting a novel therapeutic avenue for treatment of bladder pain, such as zinc supplementation.
  • Human soluble thrombomodulin-induced blockade of peripheral HMGB1-dependent allodynia in mice requires both the lectin-like and EGF-like domains., Yusuke Hayashi, Ryuichi Tsujita, Maho Tsubota, Haruka Saeki, Fumiko Sekiguchi, Goichi Honda, Atsufumi Kawabata, Biochemical and biophysical research communications, Biochemical and biophysical research communications, 495(1), 634 - 638, Jan. 01 2018 , Refereed
    Summary:Thrombomodulin (TM), an endothelial protein with anti-coagulant activity, is composed of 5 domains, D1-D5. Recombinant human soluble TM (TMα) consisting of D1-D3, which is generated in CHO cells, suppresses inflammatory and nociceptive signals by inactivating high mobility group box 1 (HMGB1), one of damage-associated molecular patterns. TMα sequesters HMGB1 with the lectin-like D1 and promotes its degradation by thrombin binding to the EGF-like D2. We prepared TM's D123, D1 and D2 by the protein expression system of yeast, and evaluated their effects on HMGB1 degradation in vitro and on the allodynia caused by HMGB1 in distinct redox forms in mice in vivo. TMα and TM's D123, but not D1, promoted the thrombin-dependent degradation of all-thiol (at-HMGB1) and disulfide HMGB1 (ds-HMGB1), an effect mimicked by TM's D2, though to a lesser extent. Intraplantar administration of TMα and TM's D123, but not D1, D2 or D1 plus D2, strongly prevented the mechanical allodynia caused by intraplantar at-HMGB1, ds-HMGB1 or lipopolysaccharide in mice. Our data suggest that, apart from the role of D3, TMα and TM's D123 require both lectin-like D1 capable of sequestering HMGB1 and EGF-like D2 responsible for thrombin-dependent degradation of HMGB1, in abolishing the allodynia caused by exogenous or endogenous HMGB1.
  • Prostanoid-dependent bladder pain caused by proteinase-activated receptor-2 activation in mice: Involvement of TRPV1 and T-type Ca2+ channels., Maho Tsubota, Tomoka Ozaki, Yuko Hayashi, Yasumasa Okawa, Ayaka Fujimura, Fumiko Sekiguchi, Hiroyuki Nishikawa, Atsufumi Kawabata, Journal of pharmacological sciences, Journal of pharmacological sciences, 136(1), 46 - 49, Jan. 2018 , Refereed
    Summary:We studied the pronociceptive role of proteinase-activated receptor-2 (PAR2) in mouse bladder. In female mice, intravesical infusion of the PAR2-activating peptide, SLIGRL-amide (SL), caused delayed mechanical hypersensitivity in the lower abdomen, namely 'referred hyperalgesia', 6-24 h after the administration. The PAR2-triggered referred hyperalgesia was prevented by indomethacin or a selective TRPV1 blocker, and restored by a T-type Ca2+ channel blocker. In human urothelial T24 cells, SL caused delayed prostaglandin E2 production and COX-2 upregulation. Our data suggest that luminal PAR2 stimulation in the bladder causes prostanoid-dependent referred hyperalgesia in mice, which involves the activation of TRPV1 and T-type Ca2+ channels.
  • Involvement of Voltage-Gated Calcium Channels in Inflammation and Inflammatory Pain., Fumiko Sekiguchi, Maho Tsubota, Atsufumi Kawabata, Biological & pharmaceutical bulletin, Biological & pharmaceutical bulletin, 41(8), 1127 - 1134, 2018 , Refereed
    Summary:Voltage-gated calcium channels (VGCCs) are classified into high-voltage-activated (HVA) channels and low-voltage-activated channels consisting of Cav3.1-3.3, known as T ("transient")-type VGCC. There is evidence that certain types of HVA channels are involved in neurogenic inflammation and inflammatory pain, in agreement with reports indicating the therapeutic effectiveness of gabapentinoids, ligands for the α2δ subunit of HVA, in treating not only neuropathic, but also inflammatory, pain. Among the Cav3 family members, Cav3.2 is abundantly expressed in the primary afferents, regulating both neuronal excitability at the peripheral terminals and spontaneous neurotransmitter release at the spinal terminals. The function and expression of Cav3.2 are modulated by a variety of inflammatory mediators including prostanoids and hydrogen sulfide (H2S), a gasotransmitter. The increased activity of Cav3.2 by H2S participates in colonic, bladder and pancreatic pain, and regulates visceral inflammation. Together, VGCCs are involved in inflammation and inflammatory pain, and Cav3.2 T-type VGCC is especially a promising therapeutic target for the treatment of visceral inflammatory pain in patients with irritable bowel syndrome, interstitial cystitis/bladder pain syndrome, pancreatitis, etc., in addition to neuropathic pain.
  • Macrophage-derived HMGB1 as a Pain Mediator in the Early Stage of Acute Pancreatitis in Mice: Targeting RAGE and CXCL12/CXCR4 Axis, Yuhei Irie, Maho Tsubota, Hiroyasu Ishikura, Fumiko Sekiguchi, Yuka Terada, Toshifumi Tsujiuchi, Keyue Liu, Masahiro Nishibori, Atsufumi Kawabata, JOURNAL OF NEUROIMMUNE PHARMACOLOGY, JOURNAL OF NEUROIMMUNE PHARMACOLOGY, 12(4), 693 - 707, Dec. 2017 , Refereed
    Summary:Extracellular high mobility group box 1 (HMGB1) activates the receptor for advanced glycation end products (RAGE) or Toll-like receptor 4 (TLR4) and forms a heterocomplex with CXCL12 that strongly activates CXCR4, promoting inflammatory and pain signals. In the present study, we investigated the role of HMGB1 in pancreatic pain accompanying cerulein-induced acute pancreatitis in mice. Abdominal referred hyperalgesia accompanying acute pancreatitis occurred within 1 h after 6 hourly injections of cerulein. The anti-HMGB1 neutralizing antibody or recombinant human soluble thrombomodulin (rhsTM), known to inactivate HMGB1, abolished the cerulein-induced referred hyperalgesia, but not pancreatitis itself. Plasma or pancreatic HMGB1 levels did not change, but macrophage infiltration into the pancreas occurred 1 h after cerulein treatment. Minocycline, a macrophage/microglia inhibitor, ethyl pyruvate that inhibits HMGB1 release from macrophages, or liposomal clodronate that depletes macrophages prevented the referred hyperalgesia, but not pancreatitis. Antagonists of RAGE or CXCR4, but not TLR4, strongly suppressed the cerulein-induced referred hyperalgesia, but not pancreatitis. Upregulation of RAGE, CXCR4 and CXCL12, but not TLR4, were detected in the pancreas 1 h after cerulein treatment. Our data suggest that HMGB1 regionally secreted by macrophages mediates pancreatic pain by targeting RAGE and CXCL12/CXCR4 axis in the early stage of acute pancreatitis.
  • Enhanced Hyperthermic Responses to Lipopolysaccharide in Mice Exposed to Repeated Cold Stress, Tomoyoshi Miyamoto, Yoshinori Funakami, Erika Kawashita, Shiori Tomita, Ai Nomura, Nanako Sugimoto, Haruka Saeki, Takaya Miyazaki, Maho Tsubota, Seiji Ichida, Atsufumi Kawabata, PHARMACOLOGY, PHARMACOLOGY, 99(3-4), 172 - 178, 2017 , Refereed
    Summary:Lipopolysaccharide (LPS) induces hyperthermia accompanied by various other systemic inflammatory symptoms. The rodents exposed to repeated cold (RC) stress according to a specific schedule are useful as experimental models for autonomic imbalance or fibromyalgia. It is now proven that RC-stressed mice exhibit tolerance to LPS, we examined thermal responses to LPS challenge in RC-stressed mice by monitoring core temperature using the telemetry system. Systemic administration of LPS caused bimodal hyperthermic responses in RC-stressed and unstressed mice. The magnitude of the LPS-induced hyperthermia was greater in RC-stressed mice than in unstressed mice. The RC stress-induced enhancement of hyperthermic responses to LPS was abolished by pretreatment with diclofenac, which is a cyclooxygenase (COX) inhibitor. LPS did not significantly increase COX-2 protein levels in the lung or hypothalamus of RC-stressed or unstressed mice. RC stress did not alter baseline serum corticosterone levels or their increases in response to LPS challenge. These results suggest that RC stress enhances the susceptibility of mice to LPS challenge, leading to greater prostanoid-dependent hyperthermia, which might contribute to tolerance to LPS in RC-stressed mice. (C) 2016 S. Karger AG, Basel.
  • Repeated Cold Stress Enhances the Acute Restraint Stress-Induced Hyperthermia in Mice, Tomoyoshi Miyamoto, Yoshinori Funakami, Erika Kawashita, Ai Nomura, Nanako Sugimoto, Haruka Saeki, Maho Tsubota, Seiji Ichida, Atsufumi Kawabata, BIOLOGICAL & PHARMACEUTICAL BULLETIN, BIOLOGICAL & PHARMACEUTICAL BULLETIN, 40(1), 11 - 16, Jan. 2017 , Refereed
    Summary:The rodents exposed to repeated cold stress according to a specific schedule, known as specific alternation of rhythm in temperature (SART), exhibit autonomic imbalance, and is now used as an experimental model of fibromyalgia. To explore the susceptibility of SART-stressed animals to novel acute stress, we tested whether exposure of mice to SART stress for 1 week alters the extent of acute restraint stress-induced hyperthermia. Mice were subjected to 7-d SART stress sessions; i.e., the mice were alternately exposed to 24 and 4 degrees C at 1-h intervals during the daytime (09:00-16:00) and kept at 4 degrees C overnight (16:00-09:00). SART-stressed and unstressed mice were exposed to acute restraint stress for 20-60 min, during which rectal temperature was monitored. Serum corticosterone levels were measured before and after 60-min exposure to restraint stress. SART stress itself did not alter the body temperature or serum corticosterone levels in mice. Acute restraint stress increased the body temperature and serum corticosterone levels, both responses being greater in SART-stressed mice than unstressed mice. The enhanced hyperthermic responses to acute restraint stress in SART-stressed mice were significantly attenuated by SR59230A, a beta(3) adrenoceptor antagonist, but unaffected by diazepam, an anxiolytic, mifepristone, a glucocorticoid receptor antagonist, or indomethacin, a cyclooxygenase inhibitor. These results suggest that SART stress enhances the susceptibility of mice to acute restraint stress, characterized by increased hyperthermia and corticosterone secretion, and that the increased hyperthermic responses to acute stress might involve accelerated activation of sympathetic beta(3) adrenoceptors, known to regulate non-shivering thermogenesis in the brown adipose tissue.
  • Tacrolimus Triggers Transient Receptor Potential Vanilloid-1-Dependent Relapse of Pancreatitis-Related Pain in Mice, Yuka Terada, Maho Tsubota, Hiiragi Sugo, Kohei Wakitani, Fumiko Sekiguchi, Kyoichi Wada, Mitsutaka Takada, Akira Oita, Atsufumi Kawabata, PHARMACOLOGY, PHARMACOLOGY, 99(5-6), 281 - 285, 2017 , Refereed
    Summary:Transient receptor potential vanilloid-1 (TRPV1) expressed in nociceptors is directly phosphorylated and activated by protein kinase C, and involved in the signaling of pancreatic pain. On the other hand, Ca(v)3.2 T-type Ca2+ channels expressed in nociceptors are functionally upregulated by phosphorylation with protein kinase A and also play a role in pancreatitis-related pain. Calcineurin, a phosphatase, negatively regulates various channel functions including TRPV1, and calcineurin inhibitor-induced pain syndrome by tacrolimus, a calcineurin inhibitor, used as an immunosuppressant, has been a clinical problem. We thus examined the effect of tacrolimus on pancreatitis-related pain in mice. Repeated treatment with cerulein caused referred hyperalgesia accompanying acute pancreatitis, which was unaffected by tacrolimus. Pancreatitis-related symptoms disappeared in 24 h, whereas the referred hyperalgesia recurred following the administration of tacrolimus, which was abolished by the blockers of TRPV1 but not T-type Ca2+ channels. Thus, tacrolimus appears to cause the TRPV1-dependent relapse of pancreatitis-related pain, suggesting the involvement of calcineurin in the termination of pancreatic pain. (C) 2017 S. Karger AG, Basel
  • Repeated Cold Stress Reduces Cyclophosphamide-Induced Cystitis/Bladder Pain and Macrophage Activity in Mice, Maho Tsubota, Tomoyoshi Miyamoto, Saki Hiruma, Haruka Saeki, Takaya Miyazaki, Fumiko Sekiguchi, Yoshinori Funakami, Atsufumi Kawabata, PHARMACOLOGY, PHARMACOLOGY, 99(5-6), 286 - 290, 2017 , Refereed
    Summary:We examined the effect of repeated cold (RC) stress on cyclophosphamide (CPA)-induced cystitis/bladder pain in mice, in relation to macrophage activity. CPA, given i.p. at 400 mg/kg, caused bladder pain symptoms accompanying cystitis in both unstressed and RC-stressed mice, which were prevented by the macrophage inhibitor minocycline. A low dose, that is, 200 mg/kg, of CPA still produced bladder pain symptoms in unstressed but not RC-stressed mice. Lipopoly-saccharide-induced cytokine production in peritoneal macrophages from RC-stressed mice was less than that from unstressed mice. Thus, RC stress appears to reduce CPA-induced bladder pain in mice, which may be associated with the decreased macrophage activity. (C) 2017 S. Karger AG, Basel
  • Therapeutic potential of RQ-00311651, a novel T-type Ca2+ channel blocker, in distinct rodent models for neuropathic and visceral pain, Fumiko Sekiguchi, Yuma Kawara, Maho Tsubota, Eri Kawakami, Tomoka Ozaki, Yudai Kawaishi, Shiori Tomita, Daiki Kanaoka, Shigeru Yoshida, Tsuyako Ohkubo, Atsufumi Kawabata, PAIN, PAIN, 157(8), 1655 - 1665, Aug. 2016 , Refereed
    Summary:T-type Ca2+ channels (T channels), particularly Ca(v)3.2 among the 3 isoforms, play a role in neuropathic and visceral pain. We thus characterized the effects of RQ-00311651 (RQ), a novel T-channel blocker, in HEK293 cells transfected with human Ca(v)3.1 or Ca(v)3.2 by electrophysiological and fluorescent Ca2+ signaling assays, and also evaluated the antiallodynic/antihyperalgesic activity of RQ in somatic, visceral, and neuropathic pain models in rodents. RQ-00311651 strongly suppressed T currents when tested at holding potentials of -65 similar to - 260 mV, but not -80 mV, in the Ca(v)3.1- or Ca(v)3.2-expressing cells. RQ-00311651 also inhibited high K+-induced Ca2+ signaling in those cells. In mice, RQ, administered intraperitoneally (i.p.) at 5 to 20 mg/kg or orally at 20 to 40 mg/kg, significantly suppressed the somatic hyperalgesia and visceral pain-like nociceptive behavior/referred hyperalgesia caused by intraplantar and intracolonic administration of NaHS or Na2S, H2S donors, respectively, which involve the enhanced activity of Ca(v)3.2 channels. RQ-00311651, given i.p. at 5 to 20 mg/kg, exhibited antiallodynic or antihyperalgesic activity in rats with spinal nerve injury-induced neuropathy or in rats and mice with paclitaxel-induced neuropathy. Oral and i.p. RQ at 10 to 20 mg/kg also suppressed the visceral nociceptive behavior and/or referred hyperalgesia accompanying cerulein-induced acute pancreatitis and cyclophosphamide-induced cystitis in mice. The analgesic and antihyperalgesic/antiallodynic doses of oral and i.p. RQ did not significantly affect the locomotor activity and motor coordination. Together, RQ is considered a state-dependent blocker of Ca(v)3.1/Ca(v)3.2 T channels and may serve as an orally available analgesic for treatment of neuropathic and inflammatory pain including distinct visceral pain with minimum central side effects.
  • Involvement of high mobility group box 1 in the development and maintenance of chemotherapy-induced peripheral neuropathy in rats, Takeshi Nishida, Maho Tsubota, Yudai Kawaishi, Hiroki Yamanishi, Natsuki Kamitani, Fumiko Sekiguchi, Hiroyasu Ishikura, Keyue Liu, Masahiro Nishibori, Atsufumi Kawabata, TOXICOLOGY, TOXICOLOGY, 365, 48 - 58, Jul. 2016 , Refereed
    Summary:Given that high mobility group box 1 (HMGB1), a nuclear protein, once released to the extracellular space, promotes nociception, we asked if inactivation of HMGB1 prevents or reverses chemotherapy-induced painful neuropathy in rats and also examined possible involvement of Toll-like receptor 4 (TLR4) and the receptor for advanced glycation endproduct (RAGE), known as targets for HMGB1. Painful neuropathy was produced by repeated i.p. administration of paclitaxel or vincristine in rats. Nociceptive threshold was determined by the paw pressure method and/or von Frey test in the hindpaw. Tissue protein levels were determined by immunoblotting. Repeated i.p. administration of the anti-HMGB1-neutralizing antibody or recombinant human soluble thrombomodulin (rhsTM), known to inactivate HMGB1, prevented the development of hyperalgesia and/or allodynia induced by paclitaxel or vincristine in rats. A single i.p. or intraplantar (i.pl.) administration of the antibody or rhsTM reversed the chemotherapy induced neuropathy. A single i.pl. administration of a TLR4 antagonist or low molecular weight heparin, known to inhibit RAGE, attenuated the hyperalgesia caused by i.pl. HMGB1 and also the chemotherapy induced painful neuropathy. Paclitaxel or vincristine treatment significantly decreased protein levels of HMGB1 in the dorsal root ganglia, but not sciatic nerves. HMGB1 thus participates in both development and maintenance of chemotherapy-induced painful neuropathy, in part through RAGE and TLR4. HMGB1 inactivation is considered useful to prevent and treat the chemotherapy-induced painful neuropathy. (C) 2016 Elsevier Ireland Ltd. All rights reserved.
  • Peripheral HMGB1-induced hyperalgesia in mice: Redox state-dependent distinct roles of RAGE and TLR4, Daichi Yamasoba, Maho Tsubota, Risa Domoto, Fumiko Sekiguchi, Hiroyuki Nishikawa, Keyue Liu, Masahiro Nishibori, Hiroyasu Ishikura, Tetsushi Yamamoto, Atsushi Taga, Atsufumi Kawabata, JOURNAL OF PHARMACOLOGICAL SCIENCES, JOURNAL OF PHARMACOLOGICAL SCIENCES, 130(2), 139 - 142, Feb. 2016 , Refereed
    Summary:Nuclear HMGB1 that contains 3 cysteine residues is acetylated and secreted to the extracellular space, promoting inflammation via multiple molecules such as RAGE and TLR4. We thus evaluated and characterized the redox state-dependent effects of peripheral HMGB1 on nociception. Intraplantar (i.pl.) administration of bovine thymus-derived HMGB1 (bt-HMGB1), all-thiol HMGB1 (at-HMGB1) or disulfide HMGB1 (ds-HMGB1) caused long-lasting mechanical hyperalgesia in mice. The hyperalgesia following i.pl. bt-HMGB1 or at-HMGB1 was attenuated by RAGE inhibitors, while the ds-HMGB1-induced hyperalgesia was abolished by a TLR4 antagonist. Thus, nociceptive processing by peripheral HMGB1 is considered dependent on its redox states. (C) 2016 Japanese Pharmacological Society. Production and hosting by Elsevier B.V.
  • Selective sensitization of C-fiber nociceptors by hydrogen sulfide, Yuka Aoki, Maho Tsubota, Yuta Nishimoto, Yumi Maeda, Fumiko Sekiguchi, Atsufumi Kawabata, JOURNAL OF PHARMACOLOGICAL SCIENCES, JOURNAL OF PHARMACOLOGICAL SCIENCES, 130(1), 38 - 41, Jan. 2016 , Refereed
    Summary:We examined the effects of intraplantar (i.pl.) administration of NaHS, an H2S donor, known to cause T-type Ca2+ channel (T-channel)-dependent mechanical hyperalgesia, on responsiveness to electric stimulation with 5, 250 and 2000 Hz sine waves (SW) that selectively excites C, A delta and A beta fibers, respectively. NaHS, given i.pl., caused behavioral hypersensitivity to SW stimulation at 5 Hz, but not 250 or 2000 Hz, in rats. NaHS also enhanced phosphorylation of spinal ERK following 5 Hz SW stimulation. Three distinct T-channel blockers abolished the NaHS-induced behavioral hypersensitivity to 5 Hz SW stimulation. Thus, H2S selectively sensitizes C-fiber nociceptors via T-channels. (C) 2016 Japanese Pharmacological Society. Production and hosting by Elsevier B.V.
  • Roles of Ca(v)3.2 and TRPA1 Channels Targeted by Hydrogen Sulfide in Pancreatic Nociceptive Processing in Mice With or Without Acute Pancreatitis, Yuka Terada, Mayuko Fujimura, Sachiyo Nishimura, Maho Tsubota, Fumiko Sekiguchi, Atsufumi Kawabata, JOURNAL OF NEUROSCIENCE RESEARCH, JOURNAL OF NEUROSCIENCE RESEARCH, 93(2), 361 - 369, Feb. 2015 , Refereed
    Summary:Hydrogen sulfide (H2S), formed by multiple enzymes, including cystathionine--lyase (CSE), targets Ca(v)3.2 T-type Ca2+ channels (T channels) and transient receptor potential ankyrin-1 (TRPA1), facilitating somatic pain. Pancreatitis-related pain also appears to involve activation of T channels by H2S formed by the upregulated CSE. Therefore, this study investigates the roles of the Ca(v)3.2 isoform and/or TRPA1 in pancreatic nociception in the absence and presence of pancreatitis. In anesthetized mice, AP18, a TRPA1 inhibitor, abolished the Fos expression in the spinal dorsal horn caused by injection of a TRPA1 agonist into the pancreatic duct. As did mibefradil, a T-channel inhibitor, in our previous report, AP18 prevented the Fos expression following ductal NaHS, an H2S donor. In the mice with cerulein-induced acute pancreatitis, the referred hyperalgesia was suppressed by NNC 55-0396 (NNC), a selective T-channel inhibitor; zinc chloride; or ascorbic acid, known to inhibit Ca(v)3.2 selectively among three T-channel isoforms; and knockdown of Ca(v)3.2. In contrast, AP18 and knockdown of TRPA1 had no significant effect on the cerulein-induced referred hyperalgesia, although they significantly potentiated the antihyperalgesic effect of NNC at a subeffective dose. TRPA1 but not Ca(v)3.2 in the dorsal root ganglia was downregulated at a protein level in mice with cerulein-induced pancreatitis. The data indicate that TRPA1 and Ca(v)3.2 mediate the exogenous H2S-induced pancreatic nociception in naive mice and suggest that, in the mice with pancreatitis, Ca(v)3.2 targeted by H2S primarily participates in the pancreatic pain, whereas TRPA1 is downregulated and plays a secondary role in pancreatic nociceptive signaling. (c) 2014 Wiley Periodicals, Inc.
  • Polaprezinc attenuates cyclophosphamide-induced cystitis and related bladder pain in mice, Masahiro Murakami-Nakayama, Maho Tsubota, Saki Hiruma, Fumiko Sekiguchi, Kenji Matsuyama, Takeshi Kimura, Masahiro Moriyama, Atsufumi Kawabata, JOURNAL OF PHARMACOLOGICAL SCIENCES, JOURNAL OF PHARMACOLOGICAL SCIENCES, 127(2), 223 - 228, Feb. 2015 , Refereed
    Summary:Ca(v)3.2 T-type Ca2+ channels targeted by H2S, a gasotransmitter, participate in cyclophosphamide-induced cystitis and bladder pain. Given that zinc selectively inhibits Ca(v)3.2 among T-channel isoforms and also exhibits antioxidant activity, we examined whether polaprezinc (zinc-L-carnosine), a medicine for peptic ulcer treatment and zinc supplementation, reveals preventive or therapeutic effects on bladder inflammation and/or pain in the mouse with cyclophosphamide-induced cystitis, a model for interstitial cystitis. Systemic administration of cyclophosphamide caused cystitis-related symptoms including increased bladder weight and vascular permeability, and histological signs of bladder edema, accompanied by bladder pain-like nociceptive behavior/referred hyperalgesia. All these symptoms were significantly attenuated by oral preadministration of polaprezinc at 400 mg/kg. The same dose of polaprezinc also prevented the increased malondialdehyde level, an indicator of lipid peroxidation, and protein upregulation of cystathionine-gamma-Iyase. an H2S-generating enzyme, but not occludin, a tight junction-related membrane protein, in the bladder tissue of cyclophosphamide-treated mice. Oral posttreatment with polaprezinc at 30-100 mg/kg reversed the nociceptive behavior/referred hyperalgesia in a dose-dependent manner without affecting the increased bladder weight. Together, our data show that zinc supplementation with polaprezinc prevents the cyclophosphamide-induced cystitis probably through the antioxidant activity, and, like T-channel blockers, reverses the established cystitis-related bladder pain in mice, suggesting novel therapeutic usefulness of polaprezinc. (C) 2015 Japanese Pharmacological Society. Production and hosting by Elsevier B.V. All rights reserved.
  • Role of Hydrogen Sulfide, a Gasotransmitter, in Colonic Pain and Inflammation, Maho Tsubota, Atsufumi Kawabata, YAKUGAKU ZASSHI-JOURNAL OF THE PHARMACEUTICAL SOCIETY OF JAPAN, YAKUGAKU ZASSHI-JOURNAL OF THE PHARMACEUTICAL SOCIETY OF JAPAN, 134(12), 1245 - 1252, Dec. 2014 , Refereed
    Summary:Hydrogen sulfide (H2S), the third known gaseous transmitter following nitric oxide and carbon monoxide, is generated by multiple enzymes including cystathionine-gamma-lyase (CSE) in vivo. We previously demonstrated that H2S activates Ca(v)3.2 T-type Ca2+ channels expressed on sensory neurons, leading to hyperalgesia and facilitation of inflammation. Here, we describe the role of H2S in processing of colonic pain and inflammation. Intracolonic (i.col.) administration of NaHS, an H2S donor, to mice evoked colonic pain-like nociceptive behavior and referred hyperalgesia accompanied by phosphorylation of ERK in the superficial layers of spinal dorsal horn, a marker for excitation of nociceptive neurons. The pronociceptive effect of NaHS was abolished by inhibitors or knockdown of Ca(v)3.2 and by an inhibitor of TRPA1, another target molecule of H2S. In rats with colitis induced by 2,4,6-trinitrobenzenesulfonic acid (TNBS), on the other hand, repeated i.col. administration of NaHS prevented colonic ulcer and inflammatory symptoms, which were inhibited by ablation of capsaicin-sensitive sensory neurons or T-type Ca2+ channel inhibitor. NaHS, given i.col., caused phosphorylation of ERK in the spinal dorsal horn of rats with TNBS-induced colitis, but not of naive rats. In TNBS-treated rats, Ca(v)3.2 was upregulated in the dorsal root ganglia, while CSE was downregulated in the colon. Taken together, these findings suggest that inhibitors of the CSE/H2S/Ca(v)3.2 or TRPA1 pathways might be useful for the treatment of colonic pain diseases such as irritable bowel syndrome, while H2S donors or Ca(v)3.2 activators might be useful for the treatment of inflammatory bowel disease including Crohn's disease.
  • Bladder pain relief by HMGB1 neutralization and soluble thrombomodulin in mice with cyclophosphamide-induced cystitis, Junichi Tanaka, Kaoru Yamaguchi, Hiroyasu Ishikura, Maho Tsubota, Fumiko Sekiguchi, Yukari Seki, Toshifumi Tsujiuchi, Akira Murai, Takehiro Umemura, Atsufumi Kawabata, NEUROPHARMACOLOGY, NEUROPHARMACOLOGY, 79, 112 - 118, Apr. 2014 , Refereed
    Summary:High mobility group box 1 (HMGB1), one of damage-associated molecular patterns (DAMPs), plays roles in not only inflammation but also processing of somatic pain. Given that no evidence for roles of HMGB1 in visceral pain signaling is available, we asked if HMGB1 participates in bladder pain accompanying cystitis caused by cyclophosphamide in mice, using the anti-HMGB1 neutralizing antibody and recombinant human soluble thrombomodulin (rhsTM) that sequesters HMGB1 and promotes its degradation by thrombin. Cyclophosphamide, administered i.p., caused bladder pain-like nociceptive behavior and referred hyperalgesia accompanying cystitis symptoms including increased bladder weight, an indicator of edema, in mice. The cyclophosphamide-induced bladder pain and referred hyperalgesia, but not increased bladder weight, were prevented by i.p. preadministration of the anti-HMGB1 neutralizing antibody or rhsTM. HMGB1, given i.p., facilitated the bladder pain and referred hyperalgesia caused by a subeffective dose of cyclophosphamide, an effect blocked by rhsTM. In the cyclophosphamide-treated mice, HMGB1 levels greatly decreased in the bladder tissue, particularly in the urothelial cells, but did not change in the plasma. Low molecular weight heparin, known to inhibit the receptor for advanced glycation end products (RAGE), but not lipopolysaccharide from Rhodobacter sphaeroides, an inhibitor of toll-like receptor 4 (TLR4), blocked the cyclophosphamide-induced bladder pain and referred hyperalgesia. Thus, our data indicate involvement of HMGB1 in the cyclophosphamide-induced bladder pain signaling, but not cystitis itself, and suggest that targeting HMGB1 with rhsTM or blocking RAGE might serve as a novel therapeutic strategy for the management of bladder pain. (C) 2013 Elsevier Ltd. All rights reserved.
  • Recombinant human soluble thrombomodulin prevents peripheral HMGB1-dependent hyperalgesia in rats, Junichi Tanaka, Yukari Seki, Hiroyasu Ishikura, Maho Tsubota, Fumiko Sekiguchi, Kaoru Yamaguchi, Akira Murai, Takehiro Umemura, Atsufumi Kawabata, British Journal of Pharmacology, British Journal of Pharmacology, 170(6), 1233 - 1241, Nov. 2013 , Refereed
    Summary:Background and Purpose High-mobility group box 1 (HMGB1), a nuclear protein, is actively or passively released during inflammation. Recombinant human soluble thrombomodulin (rhsTM), a medicine for treatment of disseminated intravascular coagulation (DIC), sequesters HMGB1 and promotes its degradation. Given evidence for involvement of HMGB1 in pain signalling, we determined if peripheral HMGB1 causes hyperalgesia, and then asked if rhsTM modulates the HMGB1-dependent hyperalgesia. Experimental Approach Mechanical nociceptive threshold and swelling in rat hindpaw were determined by the paw pressure test and by measuring paw thickness, respectively, and HMGB1 levels in rat hindpaw plantar tissue, dorsal root ganglion (DRG) and serum were determined by Western blotting or elisa. Key Results Intraplantar (i.pl.) administration of HMGB1 rapidly evoked paw swelling and gradually caused hyperalgesia in rats. Systemic administration of rhsTM abolished HMGB1-induced hyperalgesia, and partially blocked paw swelling. LPS, administered i.pl., rapidly produced mild paw swelling, and gradually caused hyperalgesia. The anti-HMGB1 neutralizing antibody abolished LPS-induced hyperalgesia, but partially inhibited paw swelling. rhsTM at a high dose, 10 mg kg-1, prevented both hyperalgesia and paw swelling caused by LPS. In contrast, rhsTM at low doses, 0.001-1 mg kg-1, abolished the LPS-induced hyperalgesia, but not paw swelling. HMGB1 levels greatly decreased in the hindpaw, but not DRG. Serum HMGB1 tended to increase after i.pl. LPS in rats pretreated with vehicle, but not rhsTM. Conclusion and Implications These data suggest that peripheral HMGB1 causes hyperalgesia, and that rhsTM abolishes HMGB1-dependent hyperalgesia, providing novel evidence for therapeutic usefulness of rhsTM as an analgesic. © 2013 The British Pharmacological Society.
  • Contribution of TRPA1 as a Downstream Signal of Proteinase-Activated Receptor-2 to Pancreatic Pain, Yuka Terada, Mayuko Fujimura, Sachiyo Nishimura, Maho Tsubota, Fumiko Sekiguchi, Hiroyuki Nishikawa, Atsufumi Kawabata, JOURNAL OF PHARMACOLOGICAL SCIENCES, JOURNAL OF PHARMACOLOGICAL SCIENCES, 123(3), 284 - 287, Nov. 2013 , Refereed
    Summary:We examined if TRPA1, like TRPV1, contributes to pancreatic nociceptor excitation following proteinase-activated receptor-2 (PAR2) stimulation and to pancreatitis-related pain in mice. A PAR2-activating peptide, infused into the pancreatic duct, caused spinal Fos expression, which was prevented by AP18, a TRPA1 inhibitor. Repeated administration of cerulein caused referred hyperalgesia accompanying pancreatitis, which was reversed by SB366791, a TRPV1 inhibitor, but not AP18. AP18, administered in combination with a subeffective dose of SB366791, significantly suppressed the referred hyperalgesia. Our findings suggest that TRPA1, like TRPV1, mediates PAR2-triggered pancreatic nociception and that TRPA1 in collaboration with TRPV1 latently contributes to pancreatitis-related pain.
  • Inhibition by Hydrogen Sulfide of Rabbit Platelet Aggregation and Calcium Mobilization, Hiroyuki Nishikawa, Hitomi Hayashi, Satoko Kubo, Maho Tsubota-Matsunami, Fumiko Sekiguchi, Atsufumi Kawabata, BIOLOGICAL & PHARMACEUTICAL BULLETIN, BIOLOGICAL & PHARMACEUTICAL BULLETIN, 36(8), 1278 - 1282, Aug. 2013 , Refereed
    Summary:Hydrogen sulfide (H2S), a gasotransmitter, plays a variety of roles in the mammalian body including the cardiovascular system. Given evidence that H2S donors including NaHS inhibit human platelet aggregation, we examined and characterized the effects of NaFIS on rabbit platelet aggregation and cytosolic Ca2+ mobilization. Rabbit platelet aggregation was determined in platelet-rich plasma (PRP) and washed platelets. Intracellular Ca2+ levels were monitored in Fura2-loaded washed platelets. NaHS prevented rabbit platelet aggregation induced by collagen or ADP, and the effective concentration range of NaHS was 0.1-0.3 mM in PRP and 1-3 mM in washed platelets. In washed platelets, NaHS attenuated cytosolic Ca2+ mobilization induced by collagen or ADP and also reduced platelet aggregation induced by ionomycin, a Ca2+ ionophore. The antiplatelet effect of NaHS was blocked by an adenylyl cyclase inhibitor and enhanced by a phosphodiesterase inhibitor. H2S thus suppresses rabbit platelet aggregation by interfering with both upstream and downstream signals of cytosolic Ca2+ mobilization in a cAMP-dependent manner.
  • AKAP-dependent sensitization of Cav3.2 channels via the EP4 receptor/cAMP pathway mediates PGE2-induced mechanical hyperalgesia, Fumiko Sekiguchi, Yuka Aoki, Maiko Nakagawa, Daiki Kanaoka, Yuta Nishimoto, Maho Tsubota-Matsunami, Rumi Yamanaka, Shigeru Yoshida, Atsufumi Kawabata, BRITISH JOURNAL OF PHARMACOLOGY, BRITISH JOURNAL OF PHARMACOLOGY, 168(3), 734 - 745, Feb. 2013 , Refereed
    Summary:Background and Purpose The Cav3.2 isoform of T-type Ca2+ channels (T channels) is sensitized by hydrogen sulfide, a pro-nociceptive gasotransmitter, and also by PKA that mediates PGE2-induced hyperalgesia. Here we examined and analysed Cav3.2 sensitization via the PGE2/cAMP pathway in NG108-15 cells that express Cav3.2 and produce cAMP in response to PGE2, and its impact on mechanical nociceptive processing in rats. Experimental Approach In NG108-15 cells and rat dorsal root ganglion (DRG) neurons, T-channel-dependent currents (T currents) were measured with the whole-cell patch-clamp technique. The molecular interaction of Cav3.2 with A-kinase anchoring protein 150 (AKAP150) and its phosphorylation were analysed by immunoprecipitation/immunoblotting in NG108-15 cells. Mechanical nociceptive threshold was determined by the paw pressure test in rats. Key Results In NG108-15 cells and/or rat DRG neurons, dibutyryl cAMP (db-cAMP) or PGE2 increased T currents, an effect blocked by AKAP St-Ht31 inhibitor peptide (AKAPI) or KT5720, a PKA inhibitor. The effect of PGE2 was abolished by RQ-00015986-00, an EP4 receptor antagonist. AKAP150 was co-immunoprecipitated with Cav3.2, regardless of stimulation with db-cAMP, and Cav3.2 was phosphorylated by db-cAMP or PGE2. In rats, intraplantar (i.pl.) administration of db-cAMP or PGE2 caused mechanical hyperalgesia, an effect suppressed by AKAPI, two distinct T-channel blockers, NNC 55-0396 and ethosuximide, or ZnCl2, known to inhibit Cav3.2 among T channels. Oral administration of RQ-00015986-00 suppressed the PGE2-induced mechanical hyperalgesia. Conclusion and Implications Our findings suggest that PGE2 causes AKAP-dependent phosphorylation and sensitization of Cav3.2 through the EP4 receptor/cAMP/PKA pathway, leading to mechanical hyperalgesia in rats.
  • Antihyperalgesic effect of buprenorphine involves nociceptin/orphanin FQ peptide-receptor activation in rats with spinal nerve injury-induced neuropathy, Tomoko Takahashi, Kazumasa Okubo, Shota Kojima, Hiroyuki Nishikawa, Motohide Takemura, Maho Tsubota-Matsunami, Fumiko Sekiguchi, Atsufumi Kawabata, Journal of Pharmacological Sciences, Journal of Pharmacological Sciences, 122(1), 51 - 54, 2013 , Refereed
    Summary:We evaluated the effect of buprenorphine, a mixed agonist for μ-opioid receptors and nociceptin/orphanin FQ peptide (NOP) receptors, in neuropathic rats, using the paw pressure test. Buprenorphine, administered i.p. at 50, but not 20, μg/kg, exhibited naloxone-reversible analgesic activity in naïve rats. In contrast, buprenorphine at 0.5-20 μg/kg produced a naloxone-sensitive antihyperalgesic effect in the L5 spinal nerve-injured neuropathic rats. Intrathecal injection of [N-Phe1]nociceptin(1-13) NH2, a NOP-receptor antagonist, reversed the effect of buprenorphine in neuropathic rats, but not in naïve rats. Together, buprenorphine suppresses neuropathic hyperalgesia by activating NOP and opioid receptors, suggesting its therapeutic usefulness in treatment of neuropathic pain. © The Japanese Pharmacological Society.
  • Involvement of the endogenous hydrogen sulfide/Cav3.2 T-type Ca2+channel pathway in cystitis-related bladder pain in mice, Maho Matsunami, Takahiro Miki, Kanae Nishiura, Yuko Hayashi, Yasumasa Okawa, Hiroyuki Nishikawa, Fumiko Sekiguchi, Lisa Kubo, Tomoka Ozaki, Toshifumi Tsujiuchi, Atsufumi Kawabata, BRITISH JOURNAL OF PHARMACOLOGY, BRITISH JOURNAL OF PHARMACOLOGY, 167(4), 917 - 928, Oct. 2012 , Refereed
    Summary:BACKGROUND AND PURPOSE Hydrogen sulfide (H2S), generated by enzymes such as cystathionine-?-lyase (CSE) from L-cysteine, facilitates pain signals by activating the Cav3.2 T-type Ca2+ channels. Here, we assessed the involvement of the CSE/H2S/Cav3.2 pathway in cystitis-related bladder pain. EXPERIMENTAL APPROACH Cystitis was induced by i.p. administration of cyclophosphamide in mice. Bladder pain-like nociceptive behaviour was observed and referred hyperalgesia was evaluated using von Frey filaments. Phosphorylation of ERK in the spinal dorsal horn was determined immunohistochemically following intravesical administration of NaHS, an H2S donor. KEY RESULTS Cyclophosphamide caused cystitis-related symptoms including increased bladder weight, accompanied by nociceptive changes (bladder pain-like nociceptive behaviour and referred hyperalgesia). Pretreatment with DL-propargylglycine, an inhibitor of CSE, abolished the nociceptive changes and partly prevented the increased bladder weight. CSE protein in the bladder was markedly up-regulated during development of cystitis. Mibefradil or NNC 550396, blockers of T-type Ca2+ channels, administered after the symptoms of cystitis appeared, reversed the nociceptive changes. Further, silencing of Cav3.2 protein by repeated intrathecal administration of mouse Cav3.2-targeting antisense oligodeoxynucleotides also significantly attenuated the nociceptive changes, but not the increased bladder weight. Finally, the number of cells staining positive for phospho-ERK was increased in the superficial layer of the L6 spinal cord after intravesical administration of NaHS, an effect inhibited by NNC 550396. CONCLUSION AND IMPLICATIONS Endogenous H2S, generated by up-regulated CSE, caused bladder pain and referred hyperalgesia through the activation of Cav3.2 channels, one of the T-type Ca2+ channels, in mice with cyclophosphamide-induced cystitis.
  • Hydrogen sulfide-induced mechanical hyperalgesia and allodynia require activation of both Cav3.2 and TRPA1 channels in mice, Kazumasa Okubo, Midori Matsumura, Yudai Kawaishi, Yuka Aoki, Maho Matsunami, Yasumasa Okawa, Fumiko Sekiguchi, Atsufumi Kawabata, BRITISH JOURNAL OF PHARMACOLOGY, BRITISH JOURNAL OF PHARMACOLOGY, 166(5), 1738 - 1743, Jul. 2012 , Refereed
    Summary:BACKGROUND AND PURPOSE Hydrogen sulfide, a gasotransmitter, facilitates somatic pain signals via activation of Cav3.2 T-type calcium channels in rats. Given evidence for the activation of transient receptor potential ankyrin-1 (TRPA1) channels by H2S, we asked whether TRPA1 channels, in addition to Cav3.2 channels, contribute to the H2S-induced mechanical hyperalgesia and allodynia in mice. EXPERIMENTAL APPROACH Mechanical hyperalgesia and allodynia were evaluated by the von Frey test in mice. Cav3.2 or TRPA1 channels in the sensory neurons were silenced by repeated intrathecal administration of antisense oligodeoxynucleotides in mice. KEY RESULTS Intraplantar administration of NaHS evoked hyperalgesia and allodynia in mice, an effect attenuated or abolished by NNC 550396 or mibefradil, T-type calcium channel blockers, and by ascorbic acid or zinc chloride, known to selectively inhibit Cav3.2 channels, out of the three isoforms of T-type calcium channels. Silencing of Cav3.2 channels in the sensory neurons also prevented the NaHS-induced hyperalgesia and allodynia in mice. The NaHS-induced hyperalgesia and allodynia in mice were significantly suppressed by AP18, a TRPA1 channel blocker, and by silencing of TRPA1 channels in the sensory neurons. CONCLUSIONS AND IMPLICATIONS Mechanical hyperalgesia and allodynia induced by NaHS/H2S required activation of both Cav3.2 and TRPA1 channels in mice.
  • Colonic Hydrogen Sulfide-Induced Visceral Pain and Referred Hyperalgesia Involve Activation of Both Ca(v)3.2 and TRPA1 Channels in Mice, Maho Tsubota-Matsunami, Yumi Noguchi, Yasumasa Okawa, Fumiko Sekiguchi, Atsufumi Kawabata, JOURNAL OF PHARMACOLOGICAL SCIENCES, JOURNAL OF PHARMACOLOGICAL SCIENCES, 119(3), 293 - 296, Jul. 2012 , Refereed
    Summary:Luminal hydrogen sulfide (H2S), a gasotransmitter, causes colonic pain / referred hyperalgesia in mice, most probably via activation of T-type Ca2+ channels. Here we analyzed the mechanisms for H2S-induced facilitation of colonic pain signals. Intracolonic administration of NaHS, an H2S donor, evoked visceral pain-like nociceptive behavior and referred hyperalgesia in mice, an effect abolished by NNC 55-0396, a selective T-type Ca2+-channel blocker, or by knockdown of Ca(v)3.2. AP18, a TRPA1 blocker, also prevented the NaHS-induced colonic pain and referred hyperalgesia. These findings demonstrate that H2S-induced colonic pain and referred hyperalgesia require activation of both Ca(v)3.2 and TRPA1 channels in mice.
  • Topical application of disodium isostearyl 2-O-L-ascorbyl phosphate, an amphiphilic ascorbic acid derivative, reduces neuropathic hyperalgesia in rats, Kazumasa Okubo, Hiroki Nakanishi, Maho Matsunami, Hiroharu Shibayama, Atsufumi Kawabata, BRITISH JOURNAL OF PHARMACOLOGY, BRITISH JOURNAL OF PHARMACOLOGY, 166(3), 1058 - 1068, Jun. 2012 , Refereed
    Summary:BACKGROUND AND PURPOSE Cav3.2 T-type calcium channels, targeted by H2S, are involved in neuropathic hyperalgesia in rats and ascorbic acid inhibits Cav3.2 channels. Therefore, we evaluated the effects of intraplantar (i.pl.) administration of ascorbic acid or topical application of disodium isostearyl 2-O-L-ascorbyl phosphate (DI-VCP), a skin-permeable ascorbate derivative on hyperalgesia induced by NaHS, an H2S donor, and on neuropathic hyperalgesia. EXPERIMENTAL APPROACH In rats mechanical hyperalgesia was evoked by i.pl. NaHS, and neuropathic hyperalgesia was induced by L5 spinal nerve cutting (L5SNC) or by repeated administration of paclitaxel, an anti-cancer drug. Dermal ascorbic acid levels were determined colorimetrically. KEY RESULTS The NaHS-evoked Cav3.2 channel-dependent hyperalgesia was inhibited by co-administered ascorbic acid. Topical application of DI-VCP, but not ascorbic acid, prevented the NaHS-evoked hyperalgesia, and also increased dermal ascorbic acid levels. Neuropathic hyperalgesia induced by L5SNC or paclitaxel was reversed by i.pl. NNC 550396, a selective T-type calcium channel blocker, ascorbic acid or DI-VCP, and by topical DI-VCP, but not by topical ascorbic acid. The effects of i.pl. ascorbic acid and topical DI-VCP in the paclitaxel-treated rats were characterized by the faster onset and greater magnitude, compared with their effects in the L5SNC rats. Dermal ascorbic acid levels in the hindpaw significantly decreased after paclitaxel treatment, but not L5SNC, which was reversed by topical DI-VCP. CONCLUSIONS AND IMPLICATIONS Ascorbic acid, known to inhibit Cav3.2 channels, suppressed neuropathic hyperalgesia. DI-VCP ointment for topical application may be of benefit in the treatment of neuropathic pain.
  • HYDROGEN SULFIDE-INDUCED COLONIC MUCOSAL CYTOPROTECTION INVOLVES T-TYPE CALCIUM CHANNEL-DEPENDENT NEURONAL EXCITATION IN RATS, M. Matsunami, S. Kirishi, T. Okui, A. Kawabata, JOURNAL OF PHYSIOLOGY AND PHARMACOLOGY, JOURNAL OF PHYSIOLOGY AND PHARMACOLOGY, 63(1), 61 - 68, Feb. 2012 , Refereed
    Summary:Hydrogen sulfide (H2S) is generated from L-cysteine by certain enzymes including cystathionine-gamma-lyase (CSE) and cystathionine-beta-synthase (CBS), and causes excitation of nociceptors mainly via activation of Ca(v)3.2 T-type Ca2+ channels in the peripheral tissue, facilitating somatic and colonic pain. Here, we investigated whether sensory nerves and Ca(v)3.2 are involved in the H2S-induced mucosal cytoprotection against 2,4,6-trinitrobenzenesulfonic acid (TNBS)-induced colitis in rats. Colitis was evaluated 3 days after intracolonic (i.e.) TNBS in the rat. Phosphorylation of ER K in the spinal dorsal horn was detected by immunohistochemistry. Protein expression of Ca(v)3.2 in the dorsal root ganglia (DRG) and of CSE and CBS in the colon was determined by Western blotting. Repeated i.e. NaHS significantly suppressed the TNBS-induced colitis in rats, an effect prevented by ablation of sensory nerves with repeated administration of capsaicin. Repeated pretreatment with T-type Ca2+ channel blockers including ethosuximide significantly reduced the protective effects of repeated i.e. NaHS in the rats with TNBS-induced colitis. A single i.e. administration of NaHS induced ethosuximide-sensitive prompt phosphorylation of ERK in the spinal dorsal horn at T13 and L6-S1 levesl in the rats 1 day or 3 days after TNBS treatment, but not in naive rats. Ca(v)3.2 protein was upreuulated in DRG 1 day after i.e. TNBS in rats, while CSE, but not CBS, protein was downregulated in the colon. Our findings suggest that luminal H2S causes excitation of sensory nerves most probably by activating Ca(v)3.2 T-type Ca2+ channels that are upregulated in the early stage of colitis, leading to colonic mucosal cytoprotection in rats.
  • Roles of the Hydrogen Sulfide/T-Type Calcium Channel System in Somatic and Visceral Pain Processing, Atsufumi Kawabata, Maho Matsunami, CELL/TISSUE INJURY AND CYTOPROTECTION/ORGANOPROTECTION IN THE GASTROINTESTINAL TRACT: MECHANISMS, PREVENTION AND TREATMENT, CELL/TISSUE INJURY AND CYTOPROTECTION/ORGANOPROTECTION IN THE GASTROINTESTINAL TRACT: MECHANISMS, PREVENTION AND TREATMENT, 30, 212 - 218, 2012 , Refereed
    Summary:Hydrogen sulfide (H2S), a gasotransmitter, is endogenously formed from L-cysteine by certain enzymes including cystathionine-gamma-lyase (CSE) in the mammalian body. H2S sensitizes Ca(v)3.2 T-type calcium channels, leading to excitation of sensory neurons followed by somatic hyperalgesia in rats and mice. The enhanced activity of the H2S/Ca(v)3.2 system is involved in the neuropathic pain/hyperalgesia induced by repeated administration of paclitaxel, an anti-cancer drug, or by spinal nerve injury. It is also noteworthy that the H2S-induced mechanical hyperalgesia requires activation of both Ca(v)3.2 and transient receptor potential A1 (TRPA1) channels in mice. H2S and Ca(v)3.2 T-type calcium channels are also involved in processing of visceral nociception including colonic, pancreatic and bladder pain. Endogenous H2S formed by upregulated CSE contributes to the pancreatitis-related pain. Further, the excitation of sensory nerves by H2S through T-type calcium channels exerts mucosal cytoprotection against colitis in rats. Together, endogenous H2S formed by CSE appears to stimulate sensory nerves by targeting Ca(v)3.2 1-type calcium channels and, in some cases, TRPA1 channels, leading to facilitation of somatic and visceral pain signals and also contributing to colonic mucosal cytoprotection. Thus, the CSE/H2S/Ca(v)3.2 system may serve as therapeutic targets for treatment of neuropathic or visceral pain and of colitis. Copyright (C) 2012 S. Karger AG, Basel
  • Inhibition of T-type calcium channels and hydrogen sulfide-forming enzyme reverses paclitaxel-evoked neuropathic hyperalgesia in rats, K. Okubo, T. Takahashi, F. Sekiguchi, D. Kanaoka, M. Matsunami, T. Ohkubo, J. Yamazaki, N. Fukushima, S. Yoshida, A. Kawabata, Neuroscience, Neuroscience, 188, 148 - 156, Aug. 11 2011
    Summary:Hydrogen sulfide (H2S), a gasotransmitter, facilitates pain sensation by targeting Cav3.2 T-type calcium channels. The H2S/Cav3.2 pathway appears to play a role in the maintenance of surgically evoked neuropathic pain. Given evidence that chemotherapy-induced neuropathic pain is blocked by ethosuximide, known to block T-type calcium channels, we examined if more selective T-type calcium channel blockers and also inhibitors of cystathionine-γ-lyase (CSE), a major H2S-forming enzyme in the peripheral tissue, are capable of reversing the neuropathic pain evoked by paclitaxel, an anti-cancer drug. It was first demonstrated that T-type calcium channel blockers, NNC 55-0396, known to inhibit Cav3.1, and mibefradil inhibited T-type currents in Cav3.2-transfected HEK293 cells. Repeated systemic administration of paclitaxel caused delayed development of mechanical hyperalgesia, which was reversed by single intraplantar administration of NNC 55-0396 or mibefradil, and by silencing of Cav3.2 by antisense oligodeoxynucleotides. Systemic administration of dl-propargylglycine and β-cyanoalanine, irreversible and reversible inhibitors of CSE, respectively, also abolished the established neuropathic hyperalgesia. In the paclitaxel-treated rats, upregulation of Cav3.2 and CSE at protein levels was not detected in the dorsal root ganglia (DRG), spinal cord or peripheral tissues including the hindpaws, whereas H2S content in hindpaw tissues was significantly elevated. Together, our study demonstrates the effectiveness of NNC 55-0396 in inhibiting Cav3.2, and then suggests that paclitaxel-evoked neuropathic pain might involve the enhanced activity of T-type calcium channels and/or CSE in rats, but not upregulation of Cav3.2 and CSE at protein levels, differing from the previous evidence for the neuropathic pain model induced by spinal nerve cutting in which Cav3.2 was dramatically upregulated in DRG. © 2011 IBRO.
  • ONO-8130, a selective prostanoid EP1 receptor antagonist, relieves bladder pain in mice with cyclophosphamide-induced cystitis, Takahiro Miki, Maho Matsunami, Saori Nakamura, Hiroki Okada, Hidekazu Matsuya, Atsufumi Kawabata, PAIN, PAIN, 152(6), 1373 - 1381, Jun. 2011 , Refereed
    Summary:Given the previous evidence for involvement of prostanoid EP1 receptors in facilitation of the bladder afferent nerve activity and micturition reflex, the present study investigated the effect of ONO-8130, a selective EP1 receptor antagonist, on cystitis-related bladder pain in mice. Cystitis in mice was produced by intraperitoneal administration of cyclophosphamide at 300 mg/kg. Bladder pain-like nociceptive behavior and referred hyperalgesia were assessed in conscious mice. Phosphorylation of extracellular signal-regulated kinase (ERK) in the L6 spinal cord was determined by immunohistochemistry in anesthetized mice. Cyclophosphamide treatment caused bladder pain-like nociceptive behavior and referred hyperalgesia accompanying cystitis symptoms, including increased bladder weight and vascular permeability and upregulation of cyclooxygenase-2 in the bladder tissue. Oral preadministration of ONO-8130 at 0.3-30 mg/kg strongly prevented both the bladder pain-like behavior and referred hyperalgesia in a dose-dependent manner, but had slight effect on the increased bladder weight and vascular permeability. Oral ONO-8130 at 30 mg/kg also reversed the established cystitis-related bladder pain. Intravesical administration of prostaglandin E-2 caused prompt phosphorylation of ERK in the L6 spinal cord, an effect blocked by ONO-8130. Our findings strongly suggest that the prostaglandin E-2/EP1 system participates in processing of cystitis-related bladder pain, and that EP1 antagonists including ONO-8130 are useful for treatment of bladder pain, particularly in interstitial cystitis. (C) 2011 International Association for the Study of Pain. Published by Elsevier B.V. All rights reserved.
  • CHELATING LUMINAL ZINC MIMICS HYDROGEN SULFIDE-EVOKED COLONIC PAIN IN MICE: POSSIBLE INVOLVEMENT OF T-TYPE CALCIUM CHANNELS, M. Matsunami, S. Kirishi, T. Okui, A. Kawabata, NEUROSCIENCE, NEUROSCIENCE, 181, 257 - 264, May 2011 , Refereed
    Summary:Luminal hydrogen sulfide (H2S) causes colonic pain and referred hyperalgesia in mice through activation of T-type Ca2+ channels. To test a hypothesis that H2S might chelate and remove endogenous Zn2+ that inhibits the Ca(v)3.2 isoform of T-type Ca2+ channels, facilitating visceral nociception, we asked if intracolonic (i.col.) administration of Zn2+ chelators mimics H2S-induced visceral nociception. Visceral nociceptive behavior and referred abdominal allodynia/hyperalgesia were determined after i.col. administration of NaHS, a donor for H2S, or Zn2+ chelators in mice. Phospholylation of extracellular signal-regulated protein kinase (ERK) in the spinal cord was analyzed by immunohistochemistry. The visceral nociceptive behavior and referred abdominal allodynia/hyperalgesia caused by i.col. NaHS in mice were abolished by i.col. preadministration of zinc chloride (ZnCl2), known to selectively inhibit Ca(v)3.2, but not Ca(v)3.1 or Ca-v,3.3, isoforms of T-type Ca2+ channels, and by i.p. preadministration of mibefradil, a pan-T-type Ca2+ channel blocker. Two distinct Zn2+ chelators, N,N,N',N'-tetrakis(2-pyridylmethyl)-ehylenediamine (TPEN) and dipicolinic acid, when administered i.col., mimicked the NaHS-evoked visceral nociceptive behavior and referred abdominal allodynia/hyperalgesia, which were inhibited by mibefradil and by NNC 55-0396, another T-type Ca2+ channel blocker. Like i.col. NaHS, i.col. TPEN caused prompt phosphorylation of ERK in the spinal dorsal horn, an effect blocked by mibefradil. Removal of luminal Zn2+ by H2S and other Zn2+ chelators thus produces colonic pain through activation of T-type Ca2+ channels, most probably of the Cav3.2 isoform. Hence, endogenous Zn2+ is considered to play a critical role in modulating visceral pain. (C) 2011 Published by Elsevier Ltd on behalf of IBRO.
  • Phosphorylation of ERK in the Spinal Dorsal Horn Following Pancreatic Pronociceptive Stimuli With Proteinase-Activated Receptor-2 Agonists and Hydrogen Sulfide in Rats: Evidence for Involvement of Distinct Mechanisms, Osamu Fukushima, Sachiyo Nishimura, Maho Matsunami, Yuka Aoki, Hiroyuki Nishikawa, Hiroyasu Ishikura, Atsufumi Kawabata, JOURNAL OF NEUROSCIENCE RESEARCH, JOURNAL OF NEUROSCIENCE RESEARCH, 88(14), 3198 - 3205, Nov. 2010 , Refereed
    Summary:Noxious stimuli cause prompt phosphorylation of extracellular signal-regulated kinase (ERK) in the spinal dorsal horn that contributes to facilitation of pain sensation and is often used as an immediate marker for excitation of spinal neurons following somatic and colonic nociception. Here we asked whether two distinct pronociceptive stimuli with proteinase-activated receptor-2 (PAR2) agonists and hydrogen sulfide (H(2)S) in the pancreas cause phosphorylation of ERK in the spinal dorsal horn and also examined involvement of their possible downstream signaling molecules, transient receptor potential vanilloid-1 (TRPV1) and T-type Ca(2+) channels, respectively. Capsaicin (a TRPV1 agonist), trypsin (an endogenous PAR2 agonist), SLIGRL-NH(2) (a PAR2-activating peptide), and NaHS (an H2S donor) were infused into the pancreatic duct in anesthetized rats, and phosphorylated ERK in the spinal cord was detected by immunohistochemistry. Intraductal administration of capsaicin and trypsin caused prompt phosphorylation of ERK in the superficial layers of T9, but not T5 or T12, spinal dorsal horn. SLIGRL-NH(2) and NaHS, administered in the same manner, also produced ERK phosphorylation in the corresponding spinal regions. Mibefradil, a T-type Ca(2+) channel blocker, abolished the phosphorylation of ERK caused by intraductal NaHS but not SLIGRL-NH(2). In contrast, capsazepine, an inhibitor of TRPV1, suppressed the phosphorylation of ERK caused by intraductal SLIGRL-NH(2) but not NaHS. Our data thus demonstrate that pancreatic pronociceptive stimuli with PAR2 agonists and H(2)S cause ERK phosphorylation in the spinal dorsal horn, through activation of TRPV1 and T-type Ca(2+) channels, respectively, and that those two pronociceptive pathways are independent of each other. (c) 2010 Wiley-Liss, Inc.
  • The proteinase/proteinase-activated receptor-2/transient receptor potential vanilloid-1 cascade impacts pancreatic pain in mice, Sachiyo Nishimura, Hiroyasu Ishikura, Maho Matsunami, Yui Shinozaki, Fumiko Sekiguchi, Mitsuhide Naruse, Taisuke Kitamura, Ryukichi Akashi, Kenji Matsumura, Atsufumi Kawabata, LIFE SCIENCES, LIFE SCIENCES, 87(19-22), 643 - 650, Nov. 2010 , Refereed
    Summary:Aims: Proteinase-activated receptor-2 (PAR2) and transient receptor potential vanilloid-1 (TRPV1) are co-localized in the primary afferents, and the trans-activation of TRPV1 by PAR2 activation is involved in processing of somatic pain. Given evidence for contribution of PAR2 to pancreatic pain, the present study aimed at clarifying the involvement of TRPV1 in processing of pancreatic pain by the proteinase/PAR2 pathway in mice. Main methods: Acute pancreatitis was created by repeated administration of cerulein in conscious mice, and the referred allodynia/hyperalgesia was assessed using von Frey filaments. Injection of PAR2 agonises into the pancreatic duct was achieved in anesthetized mice, and expression of Fos in the spinal cord was determined by immunohistochemistry. Key findings: The established referred allodynia/hyperalgesia following cerulein treatment was abolished by post-treatment with nafamostat mesilate, a proteinase inhibitor, and with capsazepine, a TRPV1 antagonist, in mice. Injection of trypsin, an endogenous PAR2 agonist, or SLIGRL-NH(2), a PAR2-activating peptide, into the pancreatic duct caused expression of Fos protein in the spinal superficial layers at T8-T10 levels in the mice. The spinal Fos expression caused by trypsin and by SLIGRL-NH(2) was partially blocked by capsazepine, the former effect abolished by nafamostat mesilate. Significance: Our data thus suggest that the proteinase/PAR2/TRPV1 cascade might impact pancreatic pain, in addition to somatic pain, and play a role in the maintenance of pancreatitis-related pain in mice. (C) 2010 Elsevier Inc. All rights reserved.
  • Rhodanese, but not cystathionine-gamma-lyase, is associated with dextran sulfate sodium-evoked colitis in mice: A sign of impaired colonic sulfide detoxification?, Eiichi Taniguchi, Maho Matsunami, Takeshi Kimura, Daiki Yonezawa, Tsuyoshi Ishiki, Fumiko Sekiguchi, Hiroyuki Nishikawa, Yuma Maeda, Hiroyasu Ishikura, Atsufumi Kawabata, TOXICOLOGY, TOXICOLOGY, 264(1-2), 96 - 103, Oct. 2009 , Refereed
    Summary:Clinical studies suggest that colonic luminal hydrogen sulfide (H(2)S), produced by sulfate-reducing bacteria or through other pathways, might be involved in the pathogenesis of inflammatory bowel disease (IBD). Nonetheless, this hypothesis has been poorly investigated by basic studies using laboratory animals. We thus focused on two enzymes, cystathionine-gamma-lyase (CSE) that generates H(2)S from L-cysteine, and rhodanese that directly or indirectly detoxifies H(2)S, particularly in relation to the colitis induced by dextran sulfate sodium (DSS) in mice. CSE was a major H(2)S-forming enzyme in colonic and renal homogenates from mice and rats, and the rhodanese activity was also detectable in both tissues. Colitis-related symptoms including decreased body weight gain, diarrhea, hematochezia and shortening of colon length were observed in the mice drinking DSS. Those symptoms were not or only slightly attenuated by repeated administration of a CSE inhibitor. CSE activity and protein levels in the colonic tissue did not notably change in the mice with colitis. In contrast, the activity and protein/mRNA levels of rhodanese in the colon, but not kidney, significantly decreased nearly in parallel with the development of colitis, followed by elevation of rhodanese activity in red blood cells (RBCs). These data show that rhodanese, but not CSE, is associated with DSS-induced colitis in mice, leading to a hypothesis that impaired detoxification of H(2)S due to down-regulation or suppression of colonic rhodanese is involved in IBD. The delayed enhancement of rhodanese activity in RBCs, a possible compensative event, might be available as a disease marker for IBD. (C) 2009 Elsevier Ireland Ltd. All rights reserved.
  • Luminal hydrogen sulfide plays a pronociceptive role in mouse colon, M. Matsunami, T. Tarui, K. Mitani, K. Nagasawa, O. Fukushima, K. Okubo, S. Yoshida, M. Takemura, A. Kawabata, GUT, GUT, 58(6), 751 - 761, Jun. 2009 , Refereed
    Summary:Objective: Given recent evidence that hydrogen sulfide (H(2)S), a gasotransmitter, promotes somatic pain through redox modulation of T-type Ca(2+) channels, the roles of colonic luminal H(2)S in visceral nociceptive processing in mice were examined. Methods: After intracolonic administration of NaHS, an H(2)S donor, visceral pain-like behaviour and referred abdominal allodynia/hyperalgesia were evaluated. Phosphorylation of extracellular signal-regulated protein kinase (ERK) in the spinal dorsal horn was determined immunohistochemically. The whole-cell recording technique was used to evaluate T-type Ca(2+) currents (T-currents) in cultured dorsal root ganglion (DRG) neurons. Results: Like capsaicin, NaHS, administered intracolonically at 0.5-5 nmol per mouse, triggered visceral nociceptive behaviour accompanied by referred allodynia/hyperalgesia in mice. Phosphorylation of ERK in the spinal dorsal horn was detected following intracolonic NaHS or capsaicin. The behavioural effects of intracolonic NaHS were abolished by a T-type channel blocker or an oxidant, but not inhibitors of L-type Ca(2+) channels or ATP-sensitive K(+) (K(ATP)) channels. Intraperitoneal NaHS at 60 mu mol/kg facilitated intracolonic capsaicin-evoked visceral nociception, an effect abolished by the T-type channel blocker, although it alone produced no behavioural effect. In DRG neurons, T-currents were enhanced by NaHS. Conclusions: These findings suggest that colonic luminal H(2)S/NaHS plays pronociceptive roles, and imply that the underlying mechanisms might involve sensitisation/activation of T-type channels probably in the primary afferents, aside from the issue of the selectivity of mibefradil.
  • Hydrogen sulfide as a novel mediator for pancreatic pain in rodents, S. Nishimura, O. Fukushima, H. Ishikura, T. Takahashi, M. Matsunami, T. Tsujiuchi, F. Sekiguchi, M. Naruse, Y. Kamanaka, A. Kawabata, GUT, GUT, 58(6), 762 - 770, Jun. 2009 , Refereed
    Summary:Objective: Hydrogen sulfide (H(2)S) is formed from L-cysteine by multiple enzymes including cystathionine-gamma-lyase (CSE) in mammals, and plays various roles in health and disease. Recently, a pronociceptive role for H(2)S in the processing of somatic pain was identified. Here, the involvement of H(2)S in pancreatic pain is examined. Methods: Anaesthetised rats or mice received an injection of NaHS, a donor for H(2)S, or capsaicin into the pancreatic duct, and the expression of spinal Fos protein was detected by immunohistochemistry. Pancreatitis was created by 6 hourly doses of caerulein in unanaesthetised mice, and pancreatitis-related allodynia/hyperalgesia was evaluated using von Frey hairs. CSE activity and protein levels in pancreatic tissues were measured using the colorimetric method and western blotting, respectively. Results: Either NaHS or capsaicin induced the expression of Fos protein in the superficial layers of the T8 and T9 spinal dorsal horn of rats or mice. The induction of Fos by NaHS but not capsaicin was abolished by mibefradil, a T-type Ca(2+) channel blocker. In conscious mice, repeated doses of caerulein produced pancreatitis accompanied by abdominal allodynia/hyperalgesia. Pretreatment with an inhibitor of CSE prevented the allodynia/hyperalgesia, but not the pancreatitis. A single dose of mibefradil reversed the established pancreatitis-related allodynia/hyperalgesia. Either the activity or protein expression of pancreatic CSE increased after the development of caerulein-induced pancreatitis in mice. Conclusions: The data suggest that pancreatic NaHS/H(2)S most probably targets T-type Ca(2+) channels, leading to nociception, and that endogenous H(2)S produced by CSE and possibly T-type Ca(2+) channels are involved in pancreatitis-related pain.
  • Hyperalgesia induced by spinal and peripheral hydrogen sulfide: Evidence for involvement of Ca(v)3.2 T-type calcium channels, Yumi Maeda, Yuka Aoki, Fumiko Sekiguchi, Maho Matsunami, Tomoko Takahashi, Hiroyuki Nishikawa, Atsufumi Kawabata, PAIN, PAIN, 142(1-2), 127 - 132, Mar. 2009 , Refereed
    Summary:Hydrogen sulfide (H2S), a gasotransmitter, facilitates membrane currents through T-type Ca2+ channels, and intraplantar (i.pl.) administration of NaHS, a donor of H2S, causes prompt hyperalgesia in rats. In this. context, we asked whether intrathecal (i.t.) administration of NaHS could mimic the hyperalgesic effect of i.pl. NaHS in rats, and then examined if Ca(v)3.2 isoform of T-type Ca2+ channels contributed to the pronociceptive effects of i.t. and i.pl. NaHS. Either i.t. or i.pl. administration of NaHS rapidly decreased nociceptive threshold in rats, as determined by the paw pressure method. The hyperalgesia caused by i.t. and i.pl. NaHS was abolished by co-administration of mibefradil, a pan-T-type Ca2+ channel inhibitor, and also Suppressed by pretreatment with i.t. and i.pl, zinc chloride, known to preferentially inhibit Ca(v)3.2 among T-type Ca2+ channel isoforms, respectively. Repeated i.t. administration of antisense oligodeoxynucleotides (ODNs) targeting rat Cav3.2, but not mismatch ODNs, caused silencing of Ca(v)3.2 protein in the dorsal root ganglia and spinal cord, and then attenuated the hyperalgesia induced by either i.t. or i.pl, NaHS. Our findings thus establish that spinal and peripheral NaHS/H2S activates or sensitizes Ca(v)3.2 T-type Ca2+ channels expressed in the primary afferents and/or spinal nociceptive neurons, leading to sensitization of nociceptive processing and hyperalgesia. (C) 2008 International Association for the Study of Pain. Published by Elsevier B.V. All rights reserved.
  • Hydrogen sulfide evokes neurite outgrowth and expression of high-voltage-activated Ca2+ currents in NG108-15 cells: involvement of T-type Ca2+ channels, Keita Nagasawa, Takeshi Tarui, Shigeru Yoshida, Fumiko Sekiguchi, Maho Matsunami, Ai Ohi, Kazuki Fukami, Seiji Ichida, Hiroyuki Nishikawa, Atsufumi Kawabata, JOURNAL OF NEUROCHEMISTRY, JOURNAL OF NEUROCHEMISTRY, 108(3), 676 - 684, Feb. 2009 , Refereed
    Summary:We investigated if stimulation of T-type Ca2+ channels with sodium hydrosulfide (NaHS), a donor of hydrogen sulfide (H2S), could cause neuronal differentiation of NG108-15 cells. Like dibutyryl cyclic AMP (db-cAMP), treatment with NaHS at 1.5-13.5 mM for 16 h enhanced neurite outgrowth in a concentration-dependent manner. Synergistic neuritogenic effect was obtained in the cells stimulated with NaHS in combination with db-cAMP at subeffective concentrations. Exposure to NaHS or db-cAMP for 2 days resulted in enhancement of expression of high-voltage-activated currents consisting of N-, P/Q-, L- and also other types, but not of T-type currents. Mibefradil, a pan-T-type channel blocker, abolished the neuritogenesis induced by NaHS, but not by db-cAMP. The NaHS-evoked neuritogenesis was also completely blocked by pretreatment with BAPTA/AM, a chelator of intracellular Ca2+, and by zinc chloride at a concentration known to selectively inhibit Ca(v)3.2 isoform of T-type Ca2+ channels, but not Ca(v)3.1 or Ca(v)3.3. Further, l-ascorbate, recently proven to selectively inhibit Ca(v)3.2, abolished the neuritogenic effect of NaHS, but not db-cAMP. Our data thus demonstrate that NaHS/H2S is a novel inducer of neuronal differentiation in NG108-15 cells, as characterized by neuritogenesis and expression of high-voltage-activated currents, and suggest the involvement of T-type Ca2+ channels, especially Ca(v)3.2.
  • Gastrointestinal roles for proteinase-activated receptors in health and disease, A. Kawabata, M. Matsunami, F. Sekiguchi, BRITISH JOURNAL OF PHARMACOLOGY, BRITISH JOURNAL OF PHARMACOLOGY, 153, S230 - S240, Mar. 2008 , Refereed
    Summary:It has been almost a decade since the molecular cloning of all four members of the proteinase-activated receptor ( PAR) family was completed. This unique family of G protein-coupled receptors (GPCRs) mediates specific cellular actions of various endogenous proteinases including thrombin, trypsin, tryptase, etc. and also certain exogenous enzymes. Increasing evidence has been clarifying the emerging roles played by PARs in health and disease. PARs, particularly PAR1 and PAR2, are distributed throughout the gastrointestinal (GI) tract, modulating various GI functions. One of the most important GI functions of PARs is regulation of exocrine secretion in the salivary glands, pancreas and GI mucosal epithelium. PARs also modulate motility of GI smooth muscle, involving multiple mechanisms. PAR2 appears to play dual roles in pancreatitis and related pain, being pro-inflammatory/pro-nociceptive and anti-inflammatory/anti-nociceptive. Similarly, dual roles for PAR1 and PAR2 have been demonstrated in mucosal inflammation/damage throughout the GI tract. There is also fundamental and clinical evidence for involvement of PAR2 in colonic pain. PARs are thus considered key molecules in regulation of GI functions and targets for development of drugs for treatment of various GI diseases.
  • The proteinase inhibitor camostat mesilate suppresses pancreatic pain in rodents, Hiroyasu Ishikura, Sachiyo Nishimura, Maho Matsunami, Toshifumi Tsujiuchi, Tsuyoshi Ishiki, Fumiko Sekiguchi, Mitsuhide Naruse, Toshio Nakatani, Yoshihisa Kamanaka, Atsufumi Kawabata, LIFE SCIENCES, LIFE SCIENCES, 80(21), 1999 - 2004, May 2007 , Refereed
    Summary:Camostat mesilate, an orally available proteinase inhibitor, is clinically used for treatment of pancreatitis. Given recent evidence that pancreatic proteinases including trypsin and/or proteinase-activated receptor-2 (PAR2) might be involved in pancreatic pain, we examined if camostat mesilate could suppress spinal Fos expression, a marker for neuronal activation, following specific application of trypsin to the pancreas, and pancreatitis-related referred allodynia. Trypsin, administered into the pancreatic duct, caused delayed expression of Fos proteins in the superficial layer of the bilateral T8 and T9 spinal dorsal horns in rats. The trypsin-induced spinal Fos expression was completely abolished by oral preadministration of camostat mesilate at 300 mg/kg. After hourly repeated (6 times in total) administration of caerulein, mice showed typical symptoms of pancreatitis, accompanied by mechanical allodynia in the upper abdomen (i.e., referred hyperalgesia/allodynia), as assessed by use of von Frey filaments. Camostat mesilate at 100-300 mg/kg, given orally twice before the 1st and 4th doses of cacrulein, abolished the pancreatitis-related abdominal allodynia, while it partially prevented the inflammatory signs. The same doses of camostat mesilate, when administered once after the final dose of caerulein, also revealed significant anti-allodynic effect. These data suggest that camostat mesilate prevents and/or depresses pancreatitis-induced pain and/or referred hyperalgesia/allodynia, in which proteinases including trypsin would play a critical role. (C) 2007 Elsevier Inc. All rights reserved.
  • Antiallodynic effect of etidronate, a bisphosphonate, in rats with adjuvant-induced arthritis: Involvement of ATP-sensitive K+ channels, Atsufumi Kawabata, Naoyuki Kawao, Yoshimi Hironaka, Tsuyoshi Ishiki, Maho Matsunami, Fumiko Sekiguchi, NEUROPHARMACOLOGY, NEUROPHARMACOLOGY, 51(2), 182 - 190, Aug. 2006 , Refereed
    Summary:Bisphosphonates, pyrophosphate analogues, known as inhibitors of bone resorption, appear to cause analgesia in certain clinical painful situations. To detect clinically relevant analgesic property of etidronate, a non-aminobisphosphonate, we examined and characterized its antiallodynic effect in the rat with adjuvant-induced arthritis, in comparison with alendronate, an aminobisphosphonate, as determined by the von Frey test. Repeated systemic administration of etidronate at 10-40 mg/kg/day suppressed the adjuvant-induced mechanical allodynia in rat hindpaw, an effect reaching a plateau in approximately 10 days. Systemic or intraplantar (i.pl.) administration of ATP-sensitive K+ (K-ATP(+)) channel inhibitors, glibenclamide and/or tolbutamide, completely reversed the antiallodynic effect of etidronate within 1 h in the arthritic rats, without affecting the nociceptive scores in naive or arthritic animals that had not received etidronate. Alendronate, administered repeatedly, also revealed similar glibenclamide-reversible antiallodynic effect. In contrast, the antiallodynic effect of repeated systemic indomethacin was resistant to i.pl. glibenclamide in the arthritic rats. Repeated administration of etidronate or alendronate only slightly attenuated the adjuvant-evoked hindpaw edema. Among K-ATP(+) channel subunits, mRNAs for Kir6.1, SUR1, SUR2A and SUR2B were abundant in rat dorsal root ganglia, while Kir6.2 mRNA was poor. Our data demonstrate that repeated etidronate as well as alendronate exhibits antiallodynic activity in arthritic rats, which might be clinically relevant, and suggest involvement of K-ATP(+) channels in the underlying mechanisms. (c) 2006 Elsevier Ltd. All rights reserved.
  • Colonic hyperalgesia triggered by proteinase-activated receptor-2 in mice: Involvement of endogenous bradykinin, A Kawabata, N Kawao, T Kitano, M Matsunami, R Satoh, T Ishiki, T Masuko, T Kanke, N Saito, NEUROSCIENCE LETTERS, NEUROSCIENCE LETTERS, 402(1-2), 167 - 172, Jul. 2006 , Refereed
    Summary:Intracolonic (i.col.) administration of the PAR2-activating peptide (PAR2AP) SLIGRL-NH2 Slowly develops visceral hypersensitivity to i.col. capsaicin in ddY mice. Thus, we further analyzed roles of PAR2 in colonic hypersensitivity, using the novel potent PAR2AP, 2-furoyl-LIGRL-NH2 and PAR2-knockout (KO) mice. In ddY mice, i.col. 2-furoyl-LIGRL-NH2 produced delayed (6 h later) facilitation of capsaicin-evoked visceral nociception, an effect,being much more potent than SLIGRL-NH2. Such effects were mimicked by i.col. trypsin. In wild-type (WT), but not PAR2-KO, mice of C57BL/6 background, i.col. PAR2 agonists caused delayed facilitation of sensitivity to capsaicin. The PAR2-triggered visceral hypersensitivity was abolished by a bradykinin B2 receptor antagonist, HOE-140. Our data thus provide ultimate evidence for role of PAR2 in colonic hypersensitivity, and suggest involvement of the bradykinin-B2 pathway. (c) 2006 Elsevier Ireland Ltd. All rights reserved.
  • Suppression of pancreatitis-related allodynia/hyperalgesia by proteinase-activated receptor-2 in mice, A Kawabata, M Matsunami, M Tsutsumi, T Ishiki, O Fukushima, F Sekiguchi, N Kawao, T Minami, T Kanke, N Saito, BRITISH JOURNAL OF PHARMACOLOGY, BRITISH JOURNAL OF PHARMACOLOGY, 148(1), 54 - 60, May 2006 , Refereed
    Summary:1 Proteinase-activated receptor-2 (PAR2), a receptor activated by trypsin and tryptase, is abundantly expressed in the gastrointestinal tract including the C-fiber terminal, and might play a role in processing of visceral pain. In the present study, we examined and characterized the roles of PAR2 in pancreatitis-related abdominal hyperalgesia/allodynia in mice. 2 Caerulein, administered i.p. once, caused a small increase in abdominal sensitivity to stimulation with von Frey hairs, without causing pancreatitis, in PAR2-knockout (KO) mice, but not wild-type (WT) mice. 3 Caerulein, given hourly six times in total, caused more profound abdominal hyperalgesia/allodynia in PAR2-KO mice, as compared with WT mice, although no significant differences were detected in the severity of pancreatitis between the KO and WT animals. 4 The PAR2-activating peptide, 2-furoyl-LIGRL-NH2, coadministered repeatedly with caerulein six times in total, abolished the caerulein-evoked abdominal hyperalgesia/allodynia in WT, but not PAR2-KO, mice. Repeated doses of 2-furoyl-LIGRL-NH2 moderately attenuated the severity of caerulein-induced pancreatitis in WT animals. 5 Our data from experiments using PAR2-KO mice provide evidence that PAR2 functions to attenuate pancreatitis-related abdominal hyperalgesia/allodynia without affecting pancreatitis itself, although the PAR2AP applied exogenously is not only antinociceptive but also anti-inflammatory.

Conference Activities & Talks

  • HMGB1 による痛みの増強に対するヒト型可溶性トロンボモジュリンの抑制作用に関与する分子メカニズム, 辻田隆一, 坪田真帆, 林佑亮, 佐伯晴香, 本田剛一, 川畑篤史, 第40回日本神経科学大会,   2017 12 14
  • A role of macrophage-derived HMGB1 in paclitaxel-induced peripheral neuropathy in mice., Domoto, R, Yamasoba, D, Yamanishi, H, Sekiguchi, F, Tsubota, M, Nishibori, M, Kawabata, A,   2016 10 27
  • Cav3.2 T-type calcium channels as therapeutic targets for the oxaliplatin-induced peripheral neuropathy., Miyazaki, T, Fukuda, R, Tsubota, M, Kawabata, A,   2016 10 27
  • Molecular mechanisms for the upregulation of Cav3.2 T-type calcium channels in the neuropathic pain., Tomita, S, Sekiguchi, F, Tsubota, M, Kawabata, A,   2016 10 27
  • Targeting HMGB1 and its downstream molecules for treatment of oxaliplatin-induced peripheral neuropathy., Tsubota, M, Fukuda, R, Miyazaki, T, Domoto, R, Kamitani, N, Nishida, T, Sekiguchi, F, Ishikura, H, Nishibori, M, Kawabata, A,   2016 10 27
  • Involvement of Cav3.2 T-type Ca2+channels in the oxaliplatin-induced neuropathic pain., Tsubota, M,   2016 10 02
  • Macrophage-derived HMGB1 participates in lipopolysaccharide-induced inflammatory hyperalgesia and paclitaxel-induced neuropathic pain in mice., Domoto, R, Yamasoba, D, Yamanishi, H, Sekiguchi, F, Tsubota, M, Nishibori, M, Kawabata, A,   2016 09 26
  • Cav3.2 T-type calcium channels contribute to oxaliplatin-induced neuropathic pain in mice., Miyazaki, T, Fukuda, R, Tsubota, M, Kawabata, A,   2016 09 26
  • Tacrolimus causes relapse of pancreatic pain through TRPV1 activation during the recovery from cerulein-induced pancreatitis in mice., Terada, Y, Tsubota, M, Sekiguchi, F, Wada, K, Kuwahara, T, Takada, M, Kawabata, A,   2016 09 26
  • High mobility group box 1 mediates substance P-induced bladder pain in mice, a model for bladder pain syndrome., Maeda, M, Irie, Y, Tsubota, M, Kubo, L, Sekiguchi, F, Ishikura, H, Nishibori, M, Kawabata, A,   2016 07 28
  • Macrophage-derived high mobility group box 1 mediates H2S-dependent bladder pain in mice with cyclophosphamide-induced cystitis., Hiramoto, S, Tsubota, M, Yamguchi, K, Tanaka, J, Sekiguchi, F, Ishikura, H, Nishibori, M, Kawabata, A,   2016 07 28
  • Involvement of macrophage-derived high mobility group box 1 in paclitaxel-induced neuropathic pain in mice., Domoto, R, Yamasoba, D, Yamanishi, H, Sekiguchi, F, Tsubota, M, Nishibori, M, Kawabata, A,   2016 07 28
  • High mobility group box 1 mediates pancreatic pain in mice., Kawabata, A, Irie, Y, Tsubota, M, Sekiguchi, F, Ishikura, H, Nishibori, M,   2016 07 28
  • Molecular mechanisms for the upregulation of Cav3.2 T-type calcium channels in the dorsal root ganglion of rats with spinal nerve injury-induced neuropathy: involvement of Egr-1 and USP5., Tomita, S, Sekiguchi, F, Tsubota, M, Kawabata, A, Neuroscience 2015,   2015 10 17
  • The electrophysiological property and antihyperalgesic activity of sophoraflavanone G and 6prenylnaringenin, novel T-type calcium channel blockers., Ono, S, Ichii, M, Yamaoka, S, Sekiguchi, F, Fujita, T, Deguchi, T, Tsubota, M, Nishikawa, H, Yoshida, S, Murata, K, Matsuda, H, Toyooka, N, Ohkubo, T, Kawabata, A, Neuroscience 2015,   2015 10 17
  • Zinc deficiency aggravates bladder pain accompanying cyclophosphamide-induced cystitis through the enhanced activity of Cav3.2 T-type Ca2+ channels in mice., Ozaki, T, Matsuoka, J, Tsubota, M, Tomita, S, Sekiguchi, F, Minami, T, Kawabata, A, Neuroscience 2015,   2015 10 17
  • Macrophage-derived high mobility group box 1 participates in the development and maintenance of pancreatic pain through the activation of RAGE and CXCR4 in mice with cerulein-induced acute pancreatitis., Irie, Y, Tsubota, M, Sekiguchi, F, Ishikura, H, Nishibori, M, Kawabata, A, Neuroscience 2015,   2015 10 17
  • Bladder pain accompanying cyclophosphamide-induced mouse cystitis involves HMGB1 release upstream of the cystathionine-gamma-lyase/H2S/Cav3.2 pathway in the bladder tissue., Kawabata, A, Tsubota, M, Yamaguchi, K, Hiramoto, S, Sekiguchi, F, Tanaka, J, Ishikura, H, Nishibori, M, Neuroscience 2015,   2015 10 17
  • Vitamin C deficiency aggravates hydrogen sulfide-induced pain/hyperalgesia and chemotherapy-induced neuropathy in mice: possible involvement of T-type calcium channels., Kawabata, A, Tsubota, M, Uebo, K, Miki, K, Sekiguchi, F, Fukuda, R, Kondo, Y, Takahashi, K, Masutomi, H, Ishigami, A,   2015 09 02
  • Involvement of high mobility group box 1 in substance P-induced cystitis-related bladder pain in mice, Irie, Y, Kubo, L, Tsubota, M, Sekiguchi, F, Ishimura, H, Nishibori, M, Kawabata, A, Pharmacology 2014,   2014 12 16
  • The NK1 receptor antagonist prevents the cyclophosphamide-induced cystitis-related bladder pain and upregulation of cystathionine-γ-lyase, an H2S-generating enzyme, in mice, Ozaki, T, Tsubota, M, Kawabata, A, Pharmacology 2014,   2014 12 16
  • Effect of tacrolimus on caerulein-induced pancreatitis-related pain in mice, Terada, Y, Tsubota, M, Sekiguchi, F, Wada, K, Kuwahara, T, Takada, M, Kawabata, K, Pharmacology 2014,   2014 12 16
  • High mobility group box 1 as a target for prevention and therapeutic treatment of chemotherapy-induced neuropathic pain., Kawabata, A, Kawaishi, Y, Nishida, T, Yamanishi, H, Kamitani, N, Tsubota, M, Sekiguchi, F, Ishikura, H, Nishibori, M, Pharmacology 2014,   2014 12 16
  • Mechanims for upregulation of cystathionine-gamma-lyase, a hydrogen sulfide-generating enzyme, in mice with cyclophosphamide-induced cystititis: Involvement of substance P/NK1 pathway and NF-kappaB signals., Ozaki, T, Tsubota, M, Kawabata, A, International Symposium “Gasotransmitters: Physiology and Pathophysiology”,   2014 09 21
  • Prevention and reversal of chemotherapy-induced neuropathic pain by HMGB1 neutralization in rodents, Kawabata, A, Kawaishi, Y, Nishida, T, Yamanishi, H, Kamitani, N, Tsubota, M, Ishikura, H, Sekiguchi, F, Nishibori, M, 9th FENS Forum of European Neuroscience,   2014 07 05
  • Ascorbic acid deficiency augments hyperalgesia induced by hydrogen sulfide in mice, Tsubota, M, Uebo, K, Miki, K, Sekiguchi, F, Kondo, Y, Takahashi, K, Masutomi, H, Ishigami, A, Kawabata, A, Third International Conference on Hydrogen Sulfide in Biology and Medicine,   2014 06 04
  • Involvement of high mobility group box 1 in cyclophosphamide-induced cystitis-related bladder pain in mice., Kawabata, A, Tanaka, J, Yamaguchi, K, Ishikura, H, Tsubota, M, Pharmacology 2013,   2013 12 17
  • Recombinant human soluble thrombomodulin abolishes bladder pain accompanying cyclophosphamide-induced cystitis in mice, Tanaka, J, Ishikura, H, Yamaguchi, K, Tsubota, M, Sekiguchi, F, Seki, Y, Kawabata, A, Neuroscience 2013,   2013 11
  • Roles of TRPA1 channels in addition to Cav3.2 or TRPV1 as the downstream signal of hydrogen sulfide or proteinase-activated receptor-2 in the development of pancreatic pain, Terada, Y, Fujimura, M, Nishimukra, S, Tsubota, M, Kawabata, A, Neuroscience 2013,   2013 11
  • RQ-00311651, a novel T-type Ca2+ channel blocker: electrophysiological characterization and anti-hyperalgesic/anti-allodynic activity in somatic and visceral pain models., Sekiguchi, F, Kawara, Y, Kanaoka, D, Kawaishi, Y, Tsubota, M, Kawakami, E, Ozaki, T, Yoshida, S, Okubo, T, Kawabata, A, Neuroscience 2013,   2013 11
  • Recombinant soluble human thrombomodulin abolishes inflammatory hyperalgesia in rats, Tanaka, J, Seki, Y, Yamaguchi, K, Ishikura, H, Tsubota, M, Sekiguchi, F, Kawabata, A, Neuroscience 2012,   2012 10 13
  • Blocking T-type calcium channels protects against the brain injury induced by middle cerebral artery occlusion and reperfusion in mice, Kawabata, A, Nishikawa, H, Matsuda, S, Fukatsu, A, Kurokawa, Y, Tsubota-Matsunami, M, Tokuyama, S, Neuroscience 2012,   2012 10 13

Misc

  • 内臓痛におけるマクロファージ由来HMGB1の役割と治療標的分子としての可能, 坪田 真帆, 川畑 篤史, PAIN RESEARCH, 34, 1, 24, 30,   2019 03
    Summary:これまでの研究により、マクロファージ由来核内蛋白high mobility group box1(HMGB1)は多様な内臓痛の発症に関与する他、癌化学療法の副作用として生じる有痛性末梢神経障害の発症に重要な役割を果たすことが明らかになっている。すなわち、HMGB1は免疫細胞と神経細胞のクロストークを媒介する分子として、難治性疼痛の発症に深く関わっていると思われる。膀胱、膵臓、大腸などにおける内臓痛へのHMGB1の関与に焦点を絞り概説した。
  • 活性硫黄種の新展開-硫化水素から過硫化物まで- 内臓痛発現における硫化水素および過硫化物の役割, 坪田 真帆, 川畑 篤史, 日本生化学会大会プログラム・講演要旨集, 91回, [1S06p, 04],   2018 09
  • 急性および慢性術後痛におけるHMGB1の役割, 川端 柚希, 林 愛理沙, 坪田 真帆, 中武 ゆい, 辻田 隆一, 関口 富美子, 西堀 正洋, 川畑 篤史, PAIN RESEARCH, 33, 2, 139, 139,   2018 06
  • 中分子ヘパリニルフェニルアラニンは化学療法誘起末梢神経障害を抑制する, 西川 裕之, 上野山 桐子, 関口 富美子, 坪田 真帆, 岡田 卓哉, 豊岡 尚樹, 川畑 篤史, PAIN RESEARCH, 33, 2, 156, 156,   2018 06
  • RAGEを標的とした化学療法誘起末梢神経障害治療薬の探索 in silicoドラッグ・リプロファイリング/リポジショニングからのアプローチ, 脇谷 航平, 関口 富美子, 坪田 真帆, 中村 真也, 仲西 功, 川畑 篤史, PAIN RESEARCH, 33, 2, 157, 157,   2018 06
  • Butyrate誘起過敏性腸症候群モデルマウスの知覚神経過敏におけるCav3.2T型Ca2+チャネルの役割, 坪田 真帆, 川畑 篤史, 潰瘍, 45, 117, 117,   2018 05
  • AzelastineはRAGEが関与する化学療法誘起末梢神経障害の発症を抑制する―ドラッグ・リプロファイリング/リポジショニング研究からの知見, 脇谷航平, 関口富美子, 坪田真帆, 中村真也, 仲西功, 川畑篤史, 日本薬理学会近畿部会プログラム・要旨集, 133rd, 44,   2018 , http://jglobal.jst.go.jp/public/201802250128640323
  • T型Ca2+チャネル阻害活性を有するホップ成分6‐prenylnaringeninは結腸痛を抑制する, 坪田真帆, 松井和樹, 中野真希, 友近拳, 関口富美子, 川畑篤史, 日本薬理学会近畿部会プログラム・要旨集, 133rd, 34,   2018 , http://jglobal.jst.go.jp/public/201802257910940137
  • がん化学療法誘起末梢神経障害モデルマウスにおける中分子ヘパリニルフェニルアラニンの抗アロディニア作用, 西川裕之, 西川裕之, 上野山桐子, 関口富美子, 坪田真帆, 岡田卓哉, 豊岡尚樹, 豊岡尚樹, 川畑篤史, 日本薬理学会近畿部会プログラム・要旨集, 133rd, 33,   2018 , http://jglobal.jst.go.jp/public/201802264948184448
  • Butyrate誘起過敏性腸症候群モデルマウスの知覚神経過敏におけるCa v3.2 T型Ca2+チャネルの役割, 坪田真帆, 川畑篤史, 日本潰瘍学会プログラム・抄録集, 45th, 101,   2017 10 20 , http://jglobal.jst.go.jp/public/201802233395721741
  • プロテアソーム阻害作用を有する多発性骨髄腫治療薬bortezomibによって誘起されるマウスの神経障害性疼痛には一次知覚神経におけるCav3.2T型Ca2+チャネルの発現量増加が関与する, 関口 富美子, 冨田 詩織, 出口 智代, 坪田 真帆, 吉田 繁, 川畑 篤史, 日本生物学的精神医学会・日本神経精神薬理学会合同年会プログラム・抄録集, 39回・47回, 187, 187,   2017 09
  • マウスにおいてHMGB1足底内投与により誘起される機械的アロディニアにはNF‐κBシグナルとマクロファージが関与する, 堂本莉紗, 中島夏奈, 関口富美子, 坪田真帆, 川畑篤史, 次世代を担う創薬・医療薬理シンポジウムプログラム・要旨集, 2017, 57,   2017 08 25 , http://jglobal.jst.go.jp/public/201702234832286886
  • Butyrate誘起過敏性腸症候群モデルマウスにおける結腸痛へのマクロファージ由来HMGB1の関与, 坪田真帆, 梶谷梨絵, 野中結, 石井優子, 関口富美子, 西堀正洋, 川畑篤史, 次世代を担う創薬・医療薬理シンポジウムプログラム・要旨集, 2017, 46,   2017 08 25 , http://jglobal.jst.go.jp/public/201702242670072548
  • Bortezomib誘起神経障害性疼痛へのマクロファージ由来HMGB1の関与, 宮崎貴也, 坪田真帆, 冨田詩織, 出口智代, 関口富美子, 西堀正洋, 川畑篤史, 次世代を担う創薬・医療薬理シンポジウムプログラム・要旨集, 2017, 63,   2017 08 25 , http://jglobal.jst.go.jp/public/201702278169871275
  • オキサリプラチン誘起神経障害性疼痛の発症メカニズムの解析:Cav3.2 T型カルシウムチャネルとHMGB1の関与, 坪田真帆, 福田亮太郎, 関口富美子, 宮崎貴也, 堂本莉紗, 安井洋樹, 山下莉加, 上田慎, 西田武司, 西田武司, 石倉宏恭, 西堀正洋, 川畑篤史, Pain Research, 32, 2, 131,   2017 06 16 , http://jglobal.jst.go.jp/public/201702276801996364
  • トロンボモジュリンアルファはトロンビン依存性にHMGB1誘起痛覚過敏を抑制する, 辻田隆一, 辻田隆一, 林佑亮, 坪田真帆, 本田剛一, 川畑篤史, Pain Research, 32, 2, 154, 154,   2017 06 16 , http://jglobal.jst.go.jp/public/201702278643432421
  • プロテアソーム阻害薬bortezomib誘起神経障害性疼痛には一次知覚神経のCav3.2T型カルシウムチャネルタンパク量増加が関与する, 関口富美子, 冨田詩織, 出口智代, 坪田真帆, 吉田繁, 川畑篤史, Pain Research, 32, 2, 154,   2017 06 16 , http://jglobal.jst.go.jp/public/201702286774040419
  • プロテアソーム阻害薬bortezomib誘起神経障害性疼痛には一次知覚神経のCaV3.2T型カルシウムチャネルタンパク量増加が関与する, 関口 富美子, 冨田 詩織, 出口 智代, 坪田 真帆, 吉田 繁, 川畑 篤史, PAIN RESEARCH, 32, 2, 154, 154,   2017 06
  • トロンボモジュリンアルファの内臓痛治療への応用, 坪田 真帆, 川畑 篤史, 潰瘍, 44, 48, 53,   2017 05
  • Protease-activated receptor (PAR), MATSUNAMI Maho, SEKIGUCHI Fumiko, KAWABATA Atsufumi, Folia pharmacologica Japonica, 128, 6, 434, 436,   2006 12 01 , http://ci.nii.ac.jp/naid/10018553553
  • Ascorbic acid deficiency augments hyperalgesia induced by hydrogen sulfide in mice, Maho Tsubota, Kenta Uebo, Koki Miki, Fumiko Sekiguchi, Yoshitaka Kondo, Keita Takahashi, Hirofumi Masutomi, Akihito Ishigami, Atsufumi Kawabata, NITRIC OXIDE-BIOLOGY AND CHEMISTRY, 39, S29, S29,   2014 05 , 10.1016/j.niox.2014.03.093
  • Tacrolimus, a calcineurin inhibitor, potentiates capsaicin-evoked colonic pain in mice, Yuka Terada, Maho Tsubota, Fumiko Sekiguchi, Kyoichi Wada, Takeshi Kuwahara, Mitsutaka Takada, Atsufumi Kawabata, JOURNAL OF PHARMACOLOGICAL SCIENCES, 124, 185P, 185P,   2014
  • Preventive or therapeutic effects of anti-HMGB1 neutralizing antibody and recombinant human soluble thrombomodulin in rats with neuropathy induced by vincristine, Takeshi Nishida, Yudai Kawaishi, Hiroki Yamanishi, Maho Tsubota, Fumiko Sekiguchi, Hiroyasu Ishikura, Masahiro Nishibori, Atsufumi Kawabata, JOURNAL OF PHARMACOLOGICAL SCIENCES, 124, 101P, 101P,   2014
  • Inactivation of HMGB1 with its neutralizaing antibody and recombinant human soluble thrombomodulin suppresses paclitaxel-induced neuropathy in rats and mice, Yudai Kawaishi, Hiroki Yamanishi, Maho Tsubota, Fumiko Sekiguchi, Takeshi Nishida, Hiroyasu Ishikura, Masahiro Nishibori, Atsufumi Kawabata, JOURNAL OF PHARMACOLOGICAL SCIENCES, 124, 154P, 154P,   2014
  • Therapeutic effect of RQ-00311651, a novel T-type Ca2+. channel blocker, on distinct visceral pain, Eri Kawakami, Maho Tsubota-Matsunami, Tomoka Ozaki, Fumiko Sekiguchi, Tadashi Inoue, Takako Okumura, Atsufumi Kawabata, JOURNAL OF PHARMACOLOGICAL SCIENCES, 121, 225P, 225P,   2013
  • Physiological and pathological roles of hydrogen sulfide : a focus on its implication in visceral pain and inflammation, Maho Tsubota, Atsufumi Kawabata, JOURNAL OF PHYSIOLOGICAL SCIENCES, 63, S39, S39,   2013
  • RQ-00311651, a novel T-type calcium channel blocker: the inhibitory effects on membrane currents or calcium influx, and analgesic activity, Yuma Kawara, Fumiko Sekiguchi, Daiki Kanaoka, Yudai Kawaishi, Maho Tsubota-Matsunami, Shigeru Yoshida, Tsuyako Ohkubo, Mio Yamasawa, Norikazu Gaja, Hiroyuki Ohshiro, Tadashi Inoue, Atsufumi Kawabata, JOURNAL OF PHARMACOLOGICAL SCIENCES, 121, 224P, 224P,   2013
  • Recombinant human soluble thrombomodulin suppresses bladder pain in mice with cyclophosphamide-induced cystitis, Kaoru Yamaguchi, Junichi Tanaka, Yukari Seki, Hiroyasu Ishikura, Maho Tsubota-Matsunami, Fumiko Sekiguchi, Atsufumi Kawabata, JOURNAL OF PHARMACOLOGICAL SCIENCES, 121, 225P, 225P,   2013
  • Recombinant human soluble thrombomodulin abolishes HMGB1-dependent inflammatory hyperalgesia in rats, Yukari Seki, Junichi Tanaka, Kaoru Yamaguchi, Hiroyasu Ishikura, Maho Tsubota-Matsunami, Fumiko Sekiguchi, Atsufumi Kawabata, JOURNAL OF PHARMACOLOGICAL SCIENCES, 121, 225P, 225P,   2013
  • Application of T-type calcium channel blockers to treatment of ischemic brain injury, Hiroyuki Nishikawa, Sachi Matsuda, Anna Fukatsu, Yuko Kurokawa, Maho Matsunami, Shogo Tokuyama, Atsufumi Kawabata, JOURNAL OF PHARMACOLOGICAL SCIENCES, 118, 142P, 142P,   2012
  • Study on molecular mechanisms for interstitial cystitis using a substance P-induced delayed bladder pain model in mice: roles of endogenous hydrogen sulfide and Ca(v)3.2 T-type Ca2+ channels, Maho Matsunami, Yasumasa Okawa, Kazumasa Okubo, Fumiko Sekiguchi, Atsufumi Kawabata, JOURNAL OF PHARMACOLOGICAL SCIENCES, 118, 193P, 193P,   2012
  • Analgesic effects of a novel amphiphilic ascorbic derivative, disodium isostearyl 2-O-L-ascorbyl phosphate, as an ointment in rats with neuropathy induced by L5 spinal nerve cutting, Kazumasa Okubo, Hiroki Nakanishi, Maho Matsunami, Hiroharu Shibayama, Atsufumi Kawabata, JOURNAL OF PHARMACOLOGICAL SCIENCES, 118, 65P, 65P,   2012
  • Distinct roles of T-type Ca2+ channels and TRPA1 channels in facilitation of mechanical pain by hydrogen sulfide in mice, Kazumasa Okubo, Midori Matsumura, Maho Matsunami, Atsufumi Kawabata, JOURNAL OF PHARMACOLOGICAL SCIENCES, 115, 199P, 199P,   2011
  • Mechanisms for mucosal protection induced by hydrogen sulfide in rats with TNBS-evoked colitis: involvement of sensory neurons and T-type Ca2+ channels, Satoko Kirishi, Maho Matsunami, Takahiro Okui, Atsufumi Kawabata, JOURNAL OF PHARMACOLOGICAL SCIENCES, 115, 217P, 217P,   2011
  • Involvement of T-type Ca2+ channels, but not TRPA1 channels, in the maintenance of bladder pain related to cyclophosphamide-induced cystitis in mice, Kanae Nishiura, Maho Matsunami, Takahiro Miki, Atsufumi Kawabata, JOURNAL OF PHARMACOLOGICAL SCIENCES, 115, 254P, 254P,   2011
  • Involvement of endogenous hydrogen sulfide and T-type calcium channels in intravesical substance P-induced cystitis and referred hyperalgesia in mice, Maho Matsunami, Yasumasa Okawa, Atsufumi Kawabata, JOURNAL OF PHARMACOLOGICAL SCIENCES, 115, 253P, 253P,   2011
  • Application of disodium isostearyl 2-O-l-ascorbyl phosphate, a novel skin-permeable prodrug of ascorbic acid, as an ointment to treatment of neuropathic pain, Kazumasa Okubo, Tomoko Takahashi, Kazuki Fukami, Maho Matsunami, Hiroharu Shibayama, Atsufumi Kawabata, JOURNAL OF PHARMACOLOGICAL SCIENCES, 112, 130P, 130P,   2010
  • Effects of ascorbic acid and a novel amphiphilic ascorbic derivative on the hydrogen sulfide-induced hyperalgesia and paclitaxel-induced neuropathic pain, Kazumasa Okubo, Tomoko Takahashi, Kazuki Fukami, Maho Matsunami, Hiroharu Shibayama, Atsufumi Kawabata, JOURNAL OF PHARMACOLOGICAL SCIENCES, 112, 56P, 56P,   2010
  • Roles of proteinases, proteinase-activated receptor-2 and transient receptor potential vanilloid-1 channels in processing of pancreatic pain, Sachiyo Nishimura, Hiroyasu Ishikura, Maho Matsunami, Yui Shinozaki, Fumiko Sekiguchi, Mitsuhide Naruse, Taisuke Kitamura, Ryukichi Akashi, Kenji Matsumura, Atsufumi Kawabata, JOURNAL OF PHARMACOLOGICAL SCIENCES, 112, 114P, 114P,   2010
  • Involvement of hydrogen sulfide-targeted T-type Ca2+ channels in neurogenic inflammation and bladder pain in mice with cyclophosphamide-induced cystitis, Kanae Nishiura, Maho Matsunami, Takahiro Miki, Atsufumi Kawabata, JOURNAL OF PHARMACOLOGICAL SCIENCES, 112, 130P, 130P,   2010
  • ONO-8130, an EP1 antagonist, strongly attenuates cystitis-related bladder pain caused by cyclophosphamide in mice, Takahiro Miki, Maho Matsunami, Hiroki Okada, Hidekazu Matsuya, Atsufumi Kawabata, JOURNAL OF PHARMACOLOGICAL SCIENCES, 112, 130P, 130P,   2010
  • Roles of hydrogen sulphide and T-type calcium channels in chemotherapy-evoked neuropathic hyperalgesia in rats, K. Okubo, M. Matsunami, T. Takahashi, A. Kawabata, EUROPEAN JOURNAL OF NEUROLOGY, 16, 87, 87,   2009 10
  • Prostaglandin EP1 receptors participate in the development of bladder pain in mice with cyclophosphamide-induced cystitis, T. Miki, M. Matsunami, Y. Hayashi, K. Akaogi, Y. Kamanaka, A. Kawabata, EUROPEAN JOURNAL OF NEUROLOGY, 16, 86, 86,   2009 10
  • Hydrogen sulphide is a key mediator for bladder pain during cyclophosphamide-induced cystitis in mice, M. Matsunami, T. Miki, Y. Hayashi, A. Kawabata, EUROPEAN JOURNAL OF NEUROLOGY, 16, 85, 85,   2009 10
  • Activation of ERK in the spinal dorsal horn neurons by distinct pro-nociceptive stimuli in the pancreas: roles of hydrogen sulphide and proteinase-activated receptor-2, S. Nishimura, O. Fukushima, M. Matsunami, H. Ishikura, A. Kawabata, EUROPEAN JOURNAL OF NEUROLOGY, 16, 86, 86,   2009 10
  • Buprenorphine strongly attenuates neuropathic pain induced by spinal nerve injury or chemotherapy in laboratory animals, A. Kawabata, T. Takahashi, K. Okubo, M. Matsunami, S. Kojima, H. Nishikawa, M. Takemura, EUROPEAN JOURNAL OF NEUROLOGY, 16, 80, 80,   2009 10
  • Peripheral hydrogen sulphide induces hypersensitivity to C-fibre-preferential sine-wave stimulation and concomitant potentiation of ERK phosphorylation in the spinal dorsal horn neurons, Y. Aoki, Y. Maeda, M. Matsunami, A. Kawabata, EUROPEAN JOURNAL OF NEUROLOGY, 16, 75, 75,   2009 10
  • Peripheral hydrogen sulfide enhances C-fiber-mediated nociception and phosphorylation of ERK in spinal dorsal horn neurons, Yuka Aoki, Yumi Maeda, Maho Matsunami, Atsufumi Kawabata, JOURNAL OF PHARMACOLOGICAL SCIENCES, 109, 222P, 222P,   2009
  • Ascorbic acid inhibits hydrogen sulfide-induced hyperalgesia and paclitaxel-induced neuropathic pain, Kazumasa Okubo, Maho Matsunami, Kimie Onchi, Atsufumi Kawabata, JOURNAL OF PHARMACOLOGICAL SCIENCES, 109, 222P, 222P,   2009
  • INVOLVEMENT OF HYDROGEN SULFIDE AND T-TYPE CALCIUM CHANNELS IN THE MAINTENANCE OF HYPERALGESIA IN RATS WITH CHEMOTHERAPY-INDUCED NEUROPATHY, Kazumasa Okubo, Maho Matsunami, Tornoko Takahashi, Atsufumi Kawabata, JOURNAL OF PHYSIOLOGICAL SCIENCES, 59, 423, 423,   2009
  • POTENT ANTI-HYPERALGESIC EFFECT OF BUPRENORPHINE IN TWO DISTINCT NEUROPATHIC PAIN MODELS INDUCED BY SPINAL NERVE INJURY AND BY PACLITAXEL, Atsufumi Kawabata, Tomoko Takahashi, Maho Matsunami, Shota Kojima, Kazumasa Okubo, Motohide Takemura, Hiroyuki Nishikawa, JOURNAL OF PHYSIOLOGICAL SCIENCES, 59, 424, 424,   2009
  • Involvement of the endogenous gaseous messenger hydrogen sulfide in cyclophosphamide-induced cystitis and concomitant bladder pain, Maho Matsunami, Takahiro Miki, Atsufumi Kawabata, JOURNAL OF PHARMACOLOGICAL SCIENCES, 109, 60P, 60P,   2009
  • Correlation of enzymes responsible for generation and detoxification of hydrogen sulfide with inflammatory bowel disease (IBD) in rodents, Eiichi Taniguchi, Daiki Yonezawa, Tsuyoshi Isiki, Maho Matsunami, Fumiko Sekiguchi, Atsufumi Kawabata, JOURNAL OF PHARMACOLOGICAL SCIENCES, 103, 66P, 66P,   2007
  • Hydrogen sulfide is a novel nociceptive messenger in processing of colonic pain, Maho Matsunami, Kazumasa Okubo, Osamu Fukushima, Motohide Takemura, Atsufumi Kawabata, JOURNAL OF PHARMACOLOGICAL SCIENCES, 103, 82P, 82P,   2007
  • Involvement of the gas messenger hydrogen sulfide in pancreatic pain, Osamu Fukushima, Sachiyo Nishimura, Hiroyasu Ishikura, Tomoko Takahashi, Maho Matsunami, Fumiko Sekiguchi, Mitsuhide Naruse, Yoshihisa Kamanaka, Atsufumi Kawabata, JOURNAL OF PHARMACOLOGICAL SCIENCES, 103, 66P, 66P,   2007
  • Antinociceptive effect of camostat mesilate on pancreatitis-related pain in rodents, S Nishimura, H Ishikura, M Matsunami, T Tsujiuchi, F Sekiguchi, N Naruse, T Nakatani, Y Kamanaka, A Kawabata, JOURNAL OF PHARMACOLOGICAL SCIENCES, 100, 156P, 156P,   2006
  • Modulation of visceral pain by PAR-2: antinociceptive/protective roles during pancreatitis and involvement in delayed colonic hyperalgesia, A Kawabata, M Matsunami, N Kawao, T Kitano, M Tsutsumi, T Ishiki, R Satoh, F Sekiguchi, T Minami, T Kanke, JOURNAL OF PHARMACOLOGICAL SCIENCES, 97, 197P, 197P,   2005

Research Grants & Projects

  • Japan Society for the Promotion of Science, Grants-in-Aid for Scientific Research Grant-in-Aid for Young Scientists (B), The functional changes of Cav3.2 T-type Ca2+ channel and its regulatory factors in a mouse model for irritable bowel syndrome, This study suggest that functional upreguation of Cav3.2 T-channel has occurred in the mice with butyrate-induced IBS, and thereby direct stimulation of TRPV1, indirect activation via PAR2 stimulation, and TRPA1 stimulation might contribute to colonic hyperalgesia. These findings demonstrate that an important suggestion for elucidating the molecular mechanism of the onset of IBS.
  • Japan Society for the Promotion of Science, Grants-in-Aid for Scientific Research Grant-in-Aid for Scientific Research (C), Prevention and treatment of chemotherapy-induced neuropathic pain and visceral pain by targeting HMBG1, a nuclear protein, HMGB1, a nuclear protein, is released into the extracellular space and plays a role as one of damage-associated molecular patterns in pathological conditions. Here we analyzed the role of HMGB1 in chemotherapy-induced neuropathic pain and visceral pain. Our data indicate that HMGB1 participates in the development and maintenance of chemotherapy-induced neuropathic pain and pancreatitis-related pain.
  • Japan Society for the Promotion of Science, Grants-in-Aid for Scientific Research Grant-in-Aid for Young Scientists (B), Analysis of the mechanisms for the bladder pain in interstitial cystitis: the role of substance P and hydrogen sulfide, This study clarified that, in the bladder tissues, substance P (SP) upregulates cystathione-γ-lyase (CSE) via NK1, and endogenous hydrogen sulfide generated by SP/NK1/CSE pathway facilitates processing of inflammation and participates in the development and maintenance of bladder pain through the activation of Cav3.2. These results suggest that the drug targeting these signaling pathway appears to serve as novel therapeutic strategy for treatment of painful bladder diseases including interstitial cystitis.
  • Japan Society for the Promotion of Science, Grants-in-Aid for Scientific Research Grant-in-Aid for Scientific Research (C), Pain regulation by PKA and calcineurin through functional modulation of T-type calcium channels, Among three isoforms of T-type calcium channels, Cav3.2 expressed abundantly in the peripheral ending of nociceptors plays a critical role in nociceptive processing. We thus examined molecular mechanisms for functional regulation of Cav3.2 by phosphorylation and dephosphorylation, and its impact on pain signals. Our data show that PKA, activated by stimulation of prostaglandin EP4 receptors in an AKAP150-dependent manner, phosphorylates and functionally upregulates Cav3.2, leading to hyperalgesia, and that calcineurin, a phosphatase, negatively regulates Cav3.2 functions.