Department of Pharmacy Lecturer
Last Updated :2023/12/05

Researcher Information

J-Global ID

Research Areas

  • Life sciences / Clinical pharmacy

Association Memberships


Published Papers

  • Atsushi Kawase; Kota Irie; Naoya Matsuda; Yuzuki Takai; Hiroaki Shimada; Masahiro Iwaki
    Molecular medicine reports 26 (5) 2022/11 
    The role of high mobility group box 1 (HMGB1) in the regulation of efflux transporters in the liver and kidney remains unclear, although it has been reported that HMGB1 can increase P‑glycoprotein (P‑gp) expression in the brain. The present study aimed to clarify the involvement of HMGB1 in the regulation of P‑gp expression in the liver and kidney of mice with lipopolysaccharide (LPS)‑induced inflammation. Mice were treated with LPS or LPS + glycyrrhizin (GL); GL is as an HMGB1 inhibitor. Subsequently, the expression levels of transporters, such as P‑gp, and HMGB1 receptors, such as toll‑like receptor (TLR)4 and receptor for advanced glycation end‑products (RAGE), were determined by quantitative PCR and LC‑MS/MS‑based targeted proteomics. For the in vitro study, HepG2 and KMRC‑1 cells were used, as was a co‑culture of KMRC‑1 and differentiated THP‑1 cells. The mRNA and protein expression levels of Mdr1a and Tlr4 in the kidneys of LPS + GL‑treated mice were significantly decreased compared with those in LPS mice. The results indicated that HMGB1 had little effect on the expression of Mdr1a and Tlr4 in the liver, since there was little change in of Mdr1a and Mdr1b expression between the LPS and LPS + GL‑treated mice. Notably, regarding MDR1 mRNA expression, KMRC‑1 cells were more responsive to LPS than HepG2 cells, and KMRC‑1 cells treated with LPS exhibited increased levels compared with control KMRC‑1 cells. In differentiated THP‑1 cells, LPS treatment decreased the mRNA expression levels of TLR4, whereas they were restored to control levels by HMGB1. In conclusion, HMGB1 in the plasma and TLR4 in macrophages may be involved in the regulation of P‑gp expression in the kidneys of inflamed mice.
  • Hiroyuki Ikuta; Hiroaki Shimada; Kenjiro Sakamoto; Rena Nakamura; Atsushi Kawase; Masahiro Iwaki
    Xenobiotica; the fate of foreign compounds in biological systems 52 (7) 1 - 28 2022/10 
    Acyl glucuronides (AGs) are known as one of the causes of idiosyncratic drug toxicity (IDT). Although AGs can be enzymatically hydrolysed by β-glucuronidase and esterase, much information on their characteristics and species differences is lacking. This study was aimed to clarify species differences in AG hydrolysis between human and rat liver microsomes (HLM and RLM).To evaluate the AG hydrolysis profile, and the contribution of β-glucuronidase and esterase towards AG hydrolysis in HLM and RLM, nonsteroidal anti-inflammatory drugs (NSAIDs) were used. AGs were incubated with 0.1 M Tris-HCl buffer (pH 7.4) and 0.3 mg/mL HLM or RLM in the absence or presence of β-glucuronidase inhibitor, D-saccharic acid 1,4-lactone (D-SL) and esterase inhibitor, phenylmethylsulfonyl fluoride (PMSF).AGs of zomepirac (ZOM-AG), mefenamic acid (MEF-AG), and etodolac (ETO-AG) showed significantly higher AG hydrolysis rates in RLM than in HLM. Esterases were found to serve as AG hydrolases dominantly in HLM, whereas both esterases and β-glucuronidase equally contribute to AG hydrolysis in RLM. However, MEF-AG and ETO-AG were hydrolysed only by β-glucuronidase.We demonstrated for the first time that the activity of AG hydrolases towards NSAID-AGs differs between humans and rats.
  • Atsushi Kawase; Ouka Takashima; Satsuki Tanaka; Hiroaki Shimada; Masahiro Iwaki
    International journal of molecular sciences 23 (15) 2022/08 
    Non-steroidal anti-inflammatory drugs (NSAIDs) such as diclofenac (DIC) frequently induce drug-induced liver injury (DILI). It is unclear whether macrophages such as M1 and M2 participate in NSAID-associated DILI; elucidating this relationship could lead to a better understanding of the detailed mechanism of DILI. We co-cultured human hepatoma HepG2 cells with M1 or M2 derived from human monocytic leukemia THP-1 cells to examine the roles of M1 and M2 in DIC-induced cytotoxicity. DIC was added to the direct or indirect co-cultures of HepG2 cells with M1 or M2 (HepG2/M1 or HepG2/M2, respectively) at cell ratios of (1:0, 1:0.1, 1:0.4, and 1:1). In both direct and indirect HepG2/M2 co-cultures (1:0.4), there was lower lactate dehydrogenase release compared with HepG2/M1 co-cultures. Other NSAIDs as well as DIC showed similar protective effects of DIC-induced cytotoxicity. There were only slight differences in mRNA levels of apoptosis- and endoplasmic reticulum stress-associated factors between M1 and M2 after DIC treatment, suggesting that other factors determined the protective effects of M2 on DIC-induced cytotoxicity. Levels of high mobility group box 1 (HMGB1) in the medium and the mRNA expression levels of HMGB1 receptors were different between M1 and M2 after DIC treatment. Increased HMGB1 concentrations and expression of toll-like receptor 2 mRNA in M1 were observed compared with M2 after DIC treatment. In conclusion, these results suggested that the HMGB1/TLR2 signaling axis can be suppressed in M2 but not M1, leading to the different roles of M1 and M2 in NSAID-induced cytotoxicity.
  • Atsushi Kawase; Momoko Hatanaka; Naoya Matsuda; Hiroaki Shimada; Masahiro Iwaki
    International journal of molecular sciences 23 (15) 2022/08 
    SLC25A39/40, involved in mitochondrial GSH (mGSH) import from the cytoplasm, is essential for protection against oxidative stress and mitochondrial dysfunction. We examined the effects of cholestasis, through bile duct ligation (BDL) and lipopolysaccharide (LPS)-induced inflammation in mice, on Slc25a39/40 expression. Additionally, we used human clear cell renal carcinoma (KMRC-1) cells to elucidate the mechanism of regulation of SLC25A39/40 expression in the kidneys after LPS treatment. BDL resulted in a decrease in Slc25a39 mRNA in the liver and a decrease in Slc25a39/40 mRNA and protein in the kidneys. Consequently, there was a significant decrease in mGSH levels in the kidneys of BDL mice compared with those in sham mice. LPS treatment resulted in increased Slc25a40 expression in the kidneys. In KMRC-1 cells, the combination treatment of LPS-RS or FPS-ZM1 with LPS suppressed the LPS-induced increase in SLC25A40, suggesting that SLC25A40 expression could be regulated by the signaling pathway via toll-like receptor 4 and the receptor for advanced glycation end products, respectively. Our findings contribute to understanding the role of mGSH in the maintenance of the mitochondrial redox state. To the best of our knowledge, this is the first study that demonstrates the changes in Slc25a39/40 expression in mice with cholestasis-associated renal injury and LPS-induced inflammation.
  • Atsushi Kawase; Rio Yamashita; Tsubasa Yoshizato; Mashiro Yoshikawa; Hiroaki Shimada; Masahiro Iwaki
    International journal of molecular sciences 23 (9) 2022/04 
    A reactive metabolite of nonsteroidal anti-inflammatory drugs (NSAIDs), acyl-β-D-glucuronide (AG), covalently binds to endogenous proteins. The covalent adduct formation of NSAIDs-AG may lead to the dysfunction of target proteins. Therefore, it is important to clarify the detailed characterization of the formation of covalent protein adducts of NSAID-AG. UDP-glucuronosyltransferase (UGT) catalyzes the conversion of NSAIDs to NSAIDs-AG. The aim of this study was to perform a quantitative analysis of the covalent adduct formation of NSAIDs-AG with UGT. Diclofenac-AG and ketoprofen-AG formed covalent adducts with organelle proteins. Next, the number of covalent adducts formed between NSAIDs-AG and UGT isoforms (UGT1A1, UGT1A9, UGT2B4, and UGT2B9) was determined. The capacity of diclofenac-AG to form covalent adducts with UGT1A9 or UGT2B7 was approximately 10 times higher than that of mefenamic acid-AG. The amounts of covalent adducts of AG of propionic acid derivative NSAIDs with UGT2B were higher than those with UGT1A. Stereoselectivity was observed upon covalent binding to UGT. A significant negative correlation between the half-lives of NSAIDs-AG in phosphate buffers and the amount of covalent adduct with UGT2B7 was observed, suggesting the more labile NSAID-AG forms higher irreversible bindings to UGT. This report provides comprehensive information on the covalent adduct formation of NSAIDs-AGs with UGT.
  • Hiroaki Shimada; Hiroyuki Ikuta; Keisuke Kumazawa; Manato Nomi; Mayumi Shiojiri; Atsushi Kawase; Masahiro Iwaki
    European journal of pharmaceutical sciences : official journal of the European Federation for Pharmaceutical Sciences 174 106193 - 106193 2022/04 
    Acyl glucuronides (AGs) are considered to cause idiosyncratic drug toxicity (IDT), and evaluating the chemical instability of AGs may be useful for predicting the IDT risk of novel drug candidates. However, AGs show variations in their chemical instability, degree of formation, and enzymatic hydrolysis. Therefore, we evaluated the degree of AG formation, enzymatic hydrolysis, and chemical instability in liver microsomes and their relationship with IDT risk. Nonsteroidal anti-inflammatory drugs (NSAIDs) were classified into three categories in terms of their IDT risk as parent drugs: safe (SA), warning (WA), and withdrawn (WDN). To evaluate the enzymatic and non-enzymatic degradation of AG, the parent drugs were incubated with rat liver microsomes in the absence or presence of AG hydrolase inhibitors. The degree of AG formation and disappearance was considered as the rate constant. For all NSAIDs investigated, the number of AGs formed notably increased following addition of AG hydrolase inhibitors. Particularly, AG was produced by WDN drugs at a lower level than that produced by WA and SA drugs in the absence of AG hydrolase inhibitors but was significantly increased after adding AG hydrolase inhibitors. The rate constants of AG formation and non-enzymatic AG disappearance did not significantly differ among the WDN, WA, and SA drugs, whereas the rate constant of enzymatic AG disappearance of WDN drugs tended to be higher than those of WA and SA drugs. In conclusion, we evaluated the enzymatic degradation and chemical instability of AG by simultaneously producing it in liver microsomes. This method enables evaluation of AG degradation without preparing AG. Moreover, we determined the relationship between enzymatic AG degradation in rat liver microsomes and IDT risk.
  • Ilex latifolia葉抽出物の食後血糖上昇抑制作用
    笹井 剛一; 島田 紘明; 上野 省一; 川瀬 篤史; 森川 敏生; 岩城 正宏
    日本薬学会年会要旨集 (公社)日本薬学会 142年会 27T - pm06S 0918-9823 2022/03
  • Ilex latifolia葉抽出物の食後血糖上昇抑制作用
    笹井 剛一; 島田 紘明; 上野 省一; 川瀬 篤史; 森川 敏生; 岩城 正宏
    日本薬学会年会要旨集 (公社)日本薬学会 142年会 27T - pm06S 0918-9823 2022/03
  • Ken-Ichi Oba; Hiroaki Shimada; Ryota Hashimoto; Atsushi Kawase; Takeo Nakanishi; Masahiro Iwaki
    Endocrine regulations 56 (1) 22 - 30 2022/02 
    Objective. Carbamazepine (CBZ), a widely used antiepileptic drug, is one major cause of the idiosyncratic liver injury along with immune reactions. Conversely, prostaglandin E2 (PGE2) demonstrates a hepatoprotective effect by regulating immune reactions and promoting liver repair in various types of liver injury. However, the amount of hepatic PGE2 during CBZ-induced liver injury remains elusive. In this study, we aimed to evaluate the hepatic PGE2 levels during CBZ-induced liver injury using a mouse model. Methods. Mice were orally administered with CBZ at a dose of 400 mg/kg for 4 days, and 800 mg/kg on the 5th day. Results. Plasma alanine transaminase (ALT) level increased in some of mice 24 h after the last CBZ administration. Although median value of hepatic PGE2 amount in the CBZ-treated mice showed same extent as vehicle-treated control mice, it exhibited significant elevated level in mice with severe liver injury presented by a plasma ALT level >1000 IU/L. According to these results, mice had a plasma ALT level >1000 IU/L were defined as responders and the others as non-responders in this study. Even though, the hepatic PGE2 levels increased in responders, the hepatic expression and enzyme activity related to PGE2 production were not upregulated when compared with vehicle-treated control mice. However, the hepatic 15-hydroxyprostaglandin dehydrogenase (15-PGDH) expression and activity decreased significantly in responders when compared with control mice. Conclusions. These results indicate that elevated hepatic PGE2 levels can be attributed to the downregulation of 15-PGDH expression under CBZ-induced liver injury.
  • Atsushi Kawase; Akira Kazaoka; Hiroaki Shimada; Masahiro Iwaki
    Brain research 1768 147581 - 147581 2021/10 
    Brain penetration of cationic drugs is an important determinant of their efficacy and side effects. However, the effects of alterations in the activity of uptake transporters in the brain under inflammatory conditions on the brain penetration of cationic drugs are not fully understood. The aim of this study was to examine changes in brain penetration of cationic drugs, including diphenhydramine (DPHM), memantine (MMT), and cimetidine (CMD), and changes in the expression of uptake transporters such as organic cation transporter (Oct) in brain microvascular endothelial cells (BMECs) under inflammatory conditions. To clarify the effects of inflammation on the brain penetration of DPHM, MMT, and CMD, we performed brain microdialysis studies in a rat model of adjuvant-induced arthritis (AA). Further, differences in transporter mRNA expression levels between BMECs from control and AA rats were evaluated. Brain microdialysis showed that the unbound brain-to-plasma partition coefficient (Kp,uu,brain) for DPHM and MMT was significantly lower in AA rats compared with control rats. OCT mRNA levels were increased and proton-coupled organic cation (H+/OC) antiporter mRNA levels were decreased in AA rats compared with control rats. Taken together, our findings suggest that inflammation decreases the brain penetration of H+/OC antiporter substrates such as DPHM and MMT.
  • Atsushi Kawase; Hideyuki Mukai; Shunsuke Tateishi; Shintaro Kuroda; Akira Kazaoka; Ryosuke Satoh; Hiroaki Shimada; Reiko Sugiura; Masahiro Iwaki
    The Journal of pharmacology and experimental therapeutics 379 (1) 53 - 63 2021/07 
    In receptor-type transcription factors-mediated cytochrome P450 (P450)s induction, few studies have attempted to clarify the roles of protein kinase N (PKN) in the transcriptional regulation of P450s. This study aimed to examine the involvement of PKN in the transcriptional regulation of P450s by receptor-type transcription factors, including the aryl hydrocarbon receptor, constitutive androstane receptor (CAR), and pregnane X receptor. The mRNA and protein levels, and metabolic activity, of P450s in the livers of wild-type (WT) and double-mutant (D) mice harboring both PKN1 kinase-negative knock-in and PKN3 knockout mutations [PKN1 T778A/T778A; PKN3 -/-] were determined following treatment with activators for receptor-type transcription factors. mRNA and protein levels, and metabolic activity, of CYP2B10 were significantly higher in D mice treated with the CAR activator phenobarbital (PB), but not with 1,4-bis((3,5-dichloropyridin-2-yl)oxy)benzene, compared with WT mice. We examined the CAR-dependent pathway regulated by PKN following PB treatment, because the extent of CYP2B10 induction in WT and D mice was notably different in response to treatment with different CAR activators. The mRNA levels of Cyp2b10 in primary hepatocytes from WT and D mice treated with PB alone or in combination with SKI-1 (a Src inhibitor), or U0126 (a MEK inhibitor), were evaluated. Treatment of hepatocytes from D mice with the combination of PB with U0126, but not SKI-1, significantly increased the mRNA levels of Cyp2b10 compared with those from the corresponding WT mice. These findings suggest that PKN may have inhibitory effects on the Src-RACK1 pathway in the CAR-mediated induction of Cyp2b10 in mice livers. Significance Statement This is the first report of involvement of PKN in the transcriptional regulation of P450s. The elucidation of mechanisms responsible for induction of P450s could help optimize the pharmacotherapy and improve drug development. We examined whether the mRNA and protein levels, and activities of P450s were altered in double-mutant mice harboring both PKN1 kinase-negative knock-in and PKN3 knockout mutations. PKN1/3 negatively regulates CAR-mediated induction of Cyp2b10 through phosphorylation of a signaling molecule in the Src-RACK1 pathway.
  • Atsushi Kawase; Miho Hirosoko; Yuka Sugihara; Yunosuke Koyama; Ayaka Fukae; Hiroaki Shimada; Masahiro Iwaki
    Pharmaceuticals (Basel, Switzerland) 14 (3) 2021/03 
    As increased expression and activities of efflux transporters (ETs) often cause drug resistance in cancers, we tried modulating ET activity in cancer cells, using scaffold proteins such as ezrin/radixin/moesin (ERM) proteins, and Na+/H+ exchanger regulatory factor-1 (NHERF1)/ERM-binding phosphoprotein of 50 kDa (EBP50). To see whether EBP50 modulated ET activities in human liver cancer HepG2 cells, we used EBP50 siRNA and a designed TAT-PDZ1 peptide. The EBP50 knockdown (EBP50KD) cells had significantly higher intracellular accumulations of Rho123 and carboxy-dichlorofluorescein (CDF), but not H33342 (i.e., the respective substrates of P-glycoprotein (P-gp), multidrug resistance-associated protein (MRP), and breast cancer resistance protein (BCRP)), compared with control HepG2, suggesting that EBP50 knockdown in HepG2 cells decreased activity of P-gp and MRP but not BCRP. Treatment with TAT-PDZ1 peptide (>1 pM) resulted in significantly higher CDF accumulation in HepG2 cells, which persisted for ≥180 min after TAT-PDZ1 peptide treatment. These results imply that EBP50 can modulate ET activities. To our knowledge, this is the first report on using a competitive peptide to modulate interactions between MRP and EBP50.
  • Atsushi Kawase; Taihei Chuma; Kota Irie; Akira Kazaoka; Asuka Kakuno; Naoya Matsuda; Hiroaki Shimada; Masahiro Iwaki
    Brain, behavior, & immunity - health 10 100188 - 100188 2021/01 
    Uptake transporters in brain microvascular endothelial cells (BMECs) are involved in the penetration of basic (cationic) drugs such as diphenhydramine (DPHM) into the brain. Lipopolysaccharide (LPS)-induced inflammation alters the expression levels and activities of uptake transporters, which change the penetration of DPHM into the brain. A brain microdialysis study showed that the unbound brain-to-plasma partition coefficient (K p,uu,brain) for DPHM in LPS rats was approximately two times higher than that in control rats. The transcellular transport of DPHM to BMECs was increased when BMECs were cultured with serum from LPS rats. Compared with control rats or BMECs, the brain uptake of DPHM in LPS rats was increased and the intracellular accumulation of DPHM was increased under a high intracellular pH in BMECs from LPS rats, respectively. Treatment of BMECs with transporter inhibitors or inflammatory cytokines had little impact on the intracellular accumulation of DPHM in BMECs. This study suggests that LPS-induced inflammation promotes unidentified proton-coupled organic cation (H+/OC) antiporters that improve the penetration of DPHM into rat brain via the blood-brain barrier.
  • Noriaki Shimada; Hiroaki Shimada; Yoshiaki Itaya; Yasuhiko Tomino
    Therapeutic apheresis and dialysis : official peer-reviewed journal of the International Society for Apheresis, the Japanese Society for Apheresis, the Japanese Society for Dialysis Therapy 25 (5) 544 - 550 2020/10 [Refereed]
    The novel coronavirus disease outbreak started in Wuhan, China in December 2019 and has since spread rapidly worldwide. As almost all patients with end-stage kidney disease have been treated with hemodialysis in Japan, they have a higher risk of infection than the healthy population. Moreover, the complications of renal failure, such as hypertension and cardiovascular diseases, appear to be a risk factor of death owing to novel coronavirus disease. The reported morbidity and mortality rates of novel coronavirus disease are significantly higher in dialysis patients than in the healthy population. No treatment for novel coronavirus disease has yet been developed; thus, countermeasures to prevent the spread of coronavirus disease in dialysis facilities must be rapidly established. The latest findings on novel coronavirus disease in patients with end-stage kidney disease and the guidelines for countermeasures against the spread of novel coronavirus disease worldwide are summarized in this review.
  • Mai Inagaki; Tomohiro Nishimura; Takeo Nakanishi; Hiroaki Shimada; Saki Noguchi; Shin-Ichi Akanuma; Masanori Tachikawa; Ken-Ichi Hosoya; Ikumi Tamai; Emi Nakashima; Masatoshi Tomi
    iScience 23 (5) 101098 - 101098 2020/05 [Refereed]
    We evaluated the contribution of organic anion transporting polypeptide 2A1 (OATP2A1/SLCO2A1), a high-affinity carrier for prostaglandins (PGs), to the parturition process. At gestational day (GD) 15.5, OATP2A1 is co-localized with 15-hydroxy-PG dehydrogenase in the mouse placental junctional zone and facilitates PG degradation by delivering PGs to the cytoplasm. Slco2a1 (+/-) females mated with Slco2a1 (-/-) males frequently showed elevated circulating progesterone at GD18.5 and delayed parturition. Progesterone receptor inhibition by RU486 treatment at GD18.5 blocked the delay of parturition. In the junctional zone, PGE2 stimulated placental lactogen II (PL-II) production, resulting in higher expression of PL-II in Slco2a1 (-/-) placenta at GD18.5. Indomethacin treatment at GD15.5 suppressed the PL-II overproduction at GD18.5 in Slco2a1 (-/-) embryo-bearing dams, which promoted progesterone withdrawal and corrected the delayed parturition. These results suggest that extracellular PGE2 reduction by OATP2A1 at mid-pregnancy would be associated with progesterone withdrawal by suppressing PL-II production, triggering parturition onset.
  • Hiroaki Shimada; Ryota Hashimoto; Aya Aoki; Saya Yamada; Ken-Ichi Oba; Atsushi Kawase; Takeo Nakanishi; Masahiro Iwaki
    Prostaglandins, leukotrienes, and essential fatty acids 155 102081 - 102081 2020/04 [Refereed]
    Prostaglandin E2 (PGE2) exhibits hepatoprotective effects against various types of liver injury. However, there is little information on the disposition of endogenous PGE2 during liver injury. In the present study, we attempted to elucidate the mechanism involved in regulating PGE2 distribution during liver injury. Carbon tetrachloride (CCl4) was used to establish a liver injury mouse model. PGE2 was measured by LC-MS/MS. The plasma and hepatic PGE2 levels were significantly increased at 6 to 48 h after CCl4 treatment. The ratio of plasma levels of 13,14-dihydro-15-ketoPGE2 (PGEM), a major PGE2 metabolite, to PGE2 decreased significantly after CCl4 treatment. PGE2 synthesis and expression of enzymes related to PGE2 production were not induced, while the activity and mRNA expression of 15-prostaglandin dehydrogenase (15-PGDH/Hpgd), a major enzyme for PGE2 inactivation, decreased significantly in the liver of CCl4-treated mice compared to that of vehicle-treated control. The plasma and hepatic PGE2 levels were negatively correlated with the hepatic mRNA expression levels of Hpgd. Although the mRNA expression of organic anion transporting polypeptide 2A1 (OATP2A1/Slco2a1), a major PGE2 transporter, was upregulated, other hepatic OATPs decreased significantly at 24 h after CCl4 treatment. Immunohistochemical analysis indicated that 15-PGDH was mainly expressed in endothelial cells and that OATP2A1 was expressed at least in endothelial cells and Kupffer cells in the liver. These results suggest that the decreased 15-PGDH expression in hepatic endothelial cells is the principal mechanism for the increase in hepatic and plasma PGE2 levels due to the CCl4-induced liver injury.
  • マウスにおけるシトクロムP450誘導過程におけるプロテインキナーゼNファミリーの役割
    川瀬 篤史; 向井 秀幸; 立石 駿介; 黒田 真太郎; 松田 尚也; 佐藤 亮介; 島田 紘明; 杉浦 麗子; 岩城 正宏
    日本薬学会年会要旨集 (公社)日本薬学会 140年会 27X - pm02 0918-9823 2020/03
  • Atsushi Kawase; Misaki Nakasaka; Hatsune Bando; Saori Yasuda; Hiroaki Shimada; Masahiro Iwaki
    Inflammation 43 (1) 85 - 94 0360-3997 2020/02 [Refereed]
    Scaffold proteins such as radixin help to modulate the plasma membrane localization and transport activity of the multidrug resistance-associated protein 2 (MRP2/ABCC2) and P-glycoprotein (P-gp/ABCB1) efflux transporters in the liver. We examined changes in radixin expression and interaction with efflux transporters in adjuvant-induced arthritic (AA) rats, an animal model of rheumatoid arthritis, as well as in human liver cancer (HepG2) cells because inflammation affects drug pharmacokinetics via the efflux transporters. The expression levels of radixin and phosphorylated radixin (p-radixin) were measured 24 h after treatment with inflammatory cytokines comprising tumor necrosis factor (TNF)-α, interleukin (IL)-1β, IL-6 or sodium nitroprusside (SNP; a nitric oxide donor). The protein levels of radixin, MRP2, and P-gp in the rat liver were next examined. We also investigated whether inflammation affected the formation of complexes between radixin and MRP2 or P-gp. The mRNA and protein levels of radixin in HepG2 cells were significantly decreased by TNF-α treatment, while minimal changes were observed after treatment with IL-1β, IL-6 or SNP. TNF-α also significantly decreased the protein levels of p-radixin, suggesting that TNF-α inhibited the activation of radixin and thereby reduced the activity of the efflux transporters. Complex formation of radixin with MRP2 and P-gp was significantly decreased in AA rats but this was reversed by prednisolone and dexamethasone treatment, indicating that decreased interactions of radixin with MRP2 and P-gp likely occur during liver inflammation. These data suggest that liver inflammation reduces radixin function by decreasing its interactions with MRP2 and P-gp.
  • Shimada, Hiroaki; Kuma, Chiaki; Iseri, Taichi; Matsumura, Shin-ichi; Kawase, Atsushi; Matsuura, Masayoshi; Iwaki, Masahiro
    The tea of Ocimum gratissimum (OG) leaves has been commonly consumed by people living in Ishigaki Island, Okinawa prefecture, Japan, and is considered to be effective for improving diabetes mellitus. In this study, we aimed to clarify the inhibitory potential of OG leaves extract (OG-ext) on gastrointestinal glucose absorption and to provide theoretical evidence for the anti-hyperglycemic effect of OG-ext. The increase of blood glucose after oral administration of alpha-starch and glucose in mice was suppressed by co-administration of OG-ext. An in vitro enzymatic assay suggested that amylase and maltase were inhibited weakly by the addition of OG-ext. In Caco-2 cells, a human intestinal epithelial model, the sodium-dependent glucose transporter (SGLT) 1-mediated uptake of fluorescence glucose analog was inhibited significantly by the addition of OG-ext in a concentration-dependent manner. These results indicate that the inhibitory effect on SGLT1 is one of the mechanisms of the anti-hyperglycemic effect of the tea of OG leaves.
  • Atsushi Kawase; Yuta Inoue; Miho Hirosoko; Yuka Sugihara; Hiroaki Shimada; Masahiro Iwaki
    Journal of pharmacy & pharmaceutical sciences : a publication of the Canadian Society for Pharmaceutical Sciences, Societe canadienne des sciences pharmaceutiques 22 (1) 576 - 584 2019 [Refereed]
    PURPOSE: The plasma membrane localization and transport activity of multidrug resistance- associated protein 2 (MRP2/ABCC2) and P-glycoprotein (P-gp/ABCB1) efflux transporters are governed by transporter-associated proteins. Phosphatidylinositol 4,5-bisphosphate (PIP2) formed by phosphatidylinositol 4-phosphate 5-kinase type 1 (PIP5K1) activates the linker function of radixin for efflux transporters. Radixin is involved in the plasma membrane localization of efflux transporters. We examined whether PIP5K1 could be a target for the modulation of transporter activities in hepatocytes and cancer cells. METHODS: The effects of PIP5K1 depletion by siRNA in mouse primary hepatocytes, PANC1 human pancreatic carcinoma cells, and HepG2 human hepatocellular carcinoma cells on the intracellular accumulation of MRP2 and P-gp substrates were examined. RESULTS: PIP5K1A depletion resulted in increased intracellular accumulation of carboxydichlorofluorescein, a MRP2 fluorescent substrate, in mouse primary hepatocytes, PANC1 cells, and HepG2 cells. In PANC1 and HepG2 cells, the transport activities of MRP2 were significantly decreased by PIP5K1C depletion. However, the transport activities of P-gp were unchanged by PIP5K1 depletion. PIP2 levels were unchanged between control and PIP5K1A- or PIP5K1C-depleted HepG2 cells. MRP2 mRNA levels showed few changes in HepG2 cells following PIP5K1A or PIP5K1C depletion. The expression of phosphorylated radixin was decreased by PIP5K1A and PIP5K1C depletion, although total radixin levels were unchanged. CONCLUSIONS: These data suggest that PIP5K1A and PIP5K1C could be target proteins for modulating MRP2 function, partly because of the resulting changes of the linker function of radixin.
  • Atsushi Kawase; Akira Kazaoka; Rei Yamamoto; Risa Minakata; Hiroaki Shimada; Masahiro Iwaki
    Journal of pharmacy & pharmaceutical sciences : a publication of the Canadian Society for Pharmaceutical Sciences, Societe canadienne des sciences pharmaceutiques 22 (1) 457 - 465 2019 [Refereed]
    PURPOSE: Bile duct ligation (BDL) in experimental animals is widely used as an animal model of liver cholestasis and fibrosis. The transcriptional process and plasma membrane localization of transporters are regulated by nuclear receptors and scaffold proteins, respectively. However, the detailed changes of these factors in the livers of BDL rats remain unclear. To clarify the effects of BDL on the levels of transporters and metabolizing enzymes, nuclear receptors, and scaffold proteins, we investigated changes in mRNA and protein levels of livers from BDL rats. METHODS: Membrane proteins and microsomes were prepared from rats with BDL. The mRNA levels of transporters and nuclear receptors in livers of control and BDL rats were examined by real-time reverse transcription polymerase chain reaction. The protein levels of transporters, metabolizing enzymes and scaffold proteins in membrane proteins and microsomes were determined by liquid chromatography-tandem mass spectrometry-based targeted proteomics. RESULTS: Mdr1a mRNA was significantly decreased at 1 and 2 weeks of BDL. The mRNA levels of MRP2 were significantly decreased. The mRNA levels of nuclear receptors were significantly decreased in livers of 1-week BDL rats. The protein levels of P-gp were significantly increased by BDL. Regarding scaffold proteins, the protein levels of ezrin, moesin and EBP50 were significantly decreased at 2 weeks of BDL. The protein levels of radixin were significantly increased at 1 week of BDL. In 1-week BDL rats, the protein levels of metabolizing enzymes such as CYP and UGT were significantly decreased. CONCLUSIONS: This study reports the comprehensive changes of transporters, metabolizing enzymes, nuclear receptors, and ezrin/radixin/moesin proteins in the livers of BDL rats. The expression levels of nuclear receptors and radixin that regulate the transcription and localization of CYP and/or transporters were decreased by BDL.
  • Kawase A; Kaneto A; Ishibashi M; Kobayashi A; Shimada H; Iwaki M
    Toxicology mechanisms and methods 1 - 30 1537-6516 2018/11 [Refereed]
  • Inagaki Mai; Nishimura Tomohiro; Nakanishi Takeo; Shimada Hiroaki; Noguchi Saki; Akanuma Shin-ichi; Tachikawa Masanori; Nakashima Emi; Hosoya Ken-ichi; Tamai Ikumi; Tomi Masatoshi
    PLACENTA 69 E86  0143-4004 2018/09 [Refereed]
  • Yoshinobu Nakamura; Takeo Nakanishi; Hiroaki Shimada; Junya Shimizu; Rika Aotani; Shio Maruyama; Kei Higuchi; Takashi Okura; Yoshiharu Deguchi; Ikumi Tamai
    The Journal of neuroscience : the official journal of the Society for Neuroscience 38 (24) 5584 - 5595 0270-6474 2018/06 [Refereed]
    Prostaglandin E2 (PGE2) in the hypothalamus is a principal mediator of the febrile response. However, the role of organic anion transporting polypeptide 2A1 (OATP2A1/SLCO2A1), a prostaglandin transporter, in facilitating this response is unknown. Here, we investigated the effect of Slco2a1 deficiency on the body core temperature (Tc) and on the PGE2 concentration in hypothalamus interstitial fluid (Cisf) and CSF (Ccsf) of lipopolysaccharide (LPS; 100 μg/kg, i.p.)-treated mice of both sexes. Slco2a1-/- mice did not develop a febrile response. Ccsf was increased in Slco2a1+/+ and Slco2a1-/- mice, and Ccsf of Slco2a1-/- mice was well maintained at 5 h after LPS injection (1160 pg/ml) compared with Slco2a1+/+ mice (316 pg/ml). A microdialysis study revealed that Cisf peaked at 2 h after LPS injection in Slco2a1+/+ mice (841 pg/ml), whereas the increase in Cisf was negligible in Slco2a1-/- mice. The PGE2 plasma concentration in Slco2a1-/- mice (201 pg/ml) was significantly higher than that in Slco2a1+/+ mice (54 pg/ml) at 1 h after LPS injection, whereas the two groups showed similar PGE2 concentrations in the hypothalamus. Strong Oatp2a1 immunoreactivity was observed in F4/80-positive microglia and perivascular cells and in brain capillary endothelial cells. The changes in Tc and Cisf seen in LPS-injected Slco2a1+/+ mice were partially attenuated in monocyte-/macrophage-specific Slco2a1-/- (Slco2a1Fl/Fl/LysMCre/+) mice. Thus, OATP2A1 facilitates the LPS-induced febrile response by maintaining a high level of Cisf, possibly by regulating PGE2 secretion from F4/80-positive glial cells and/or facilitating PGE2 transport across the blood-brain barrier. These findings suggest that OATP2A1 is a useful therapeutic target for neuroinflammation.SIGNIFICANCE STATEMENT Fever is a physiological response caused by pyrogen-induced release of prostaglandin E2 (PGE2) in the hypothalamus, which plays a central role in regulating the set-point of body temperature. However, it is unclear whether the prostaglandin transporter OATP2A1/SLCO2A1 is involved in this response. We show here that LPS-induced fever is associated with increased PGE2 concentration in hypothalamus interstitial fluid (Cisf), but not in CSF (Ccsf), by means of a microdialysis study in global Slco2a1-knock-out mice and monocyte-/macrophage-specific Slco2a1-knock-out mice. The results suggest that OATP2A1 serves as a regulator of Cisf in F4/80-positive glial cells. OATP2A1 was detected immunohistochemically in brain capillary endothelial cells and, therefore, may also play a role in PGE2 transport across the blood-brain barrier.
  • Hiroaki Shimada; Yuri Kobayashi; Sakiko Tanahashi; Atsushi Kawase; Taro Ogiso; Masahiro Iwaki
    European Journal of Pharmaceutical Sciences Elsevier B.V. 112 132 - 138 1879-0720 2018/01 [Refereed]
    Nonsteroidal anti-inflammatory drugs (NSAIDs) can cause idiosyncratic liver injury. Mechanisms involved in NSAID-induced liver injury are complex. Previous studies have suggested that acyl glucuronide of NSAIDs (NSAIDs-Glu) plays an important role in the development of liver injury via covalently binds to proteins and the resultant adduct induces immunological toxicity. As only some NSAIDs-Glu are commercially available, the evaluation of covalent protein adduct formation using ready-made NSAIDs-Glu is difficult and inconvenient. Moreover, glucuronidation potency varies with the NSAID, including stereoisomers. Therefore, in this study, we simultaneously examined the glucuronidation and covalent adduct formation using enantiomers of parent NSAIDs (ibuprofen, naproxen, pranoprofen, ketoprofen, and flurbiprofen) in rat liver microsomes. Glucuronides and covalent adducts were quantified by HPLC. The amount of covalent adduct increased with NSAIDs-Glu formation in the rat liver microsomes in a time-dependent manner. A significant positive correlation was observed between the AUC of NSAIDs-Glu and that of covalent adduct, except ketoprofen. Although ketoprofen exhibited the highest glucuronidation rate among the NSAIDs investigated, the amount of covalent adduct was similar to that for pranoprofen, which had the lowest glucuronidation rate. Thus, it may be difficult for ketoprofen glucuronide to covalently bind with proteins in the rat liver microsomes. Our results suggested that the amount of glucuronide formed is a key factor in predicting covalent bond formation with protein in NSAIDs, in addition to degradability and bindability with proteins of NSAIDs-Glu. Further studies are required to confirm the relationship between the tendency of glucuronidation and the formation of covalent adducts of NSAIDs.
  • 各世代のとろみ調整剤が血糖値に与える影響
    松浦正佳; 島田紘明; 岸本理咲; 藤本和佳; 大鳥徹; 川瀬篤史; 岩城正宏
    薬局薬学 10 131 - 139 2018 [Refereed]
  • Shimada, Hiroaki; Urabe, Yuichi; Okamoto, Yuhei; Li, Zheng; Kawase, Atsushi; Morikawa, Toshio; Tu, Pengfei; Muraoka, Osamu; Iwaki, Masahiro
    JOURNAL OF FUNCTIONAL FOODS ELSEVIER SCIENCE BV 39 91 - 95 1756-4646 2017/12 [Refereed]
    Echinacoside (ECH) and acteoside (ACT), the major constituents of Cistanche tubulosa, suppress the increase in postprandial blood glucose level. Although ECH and ACT have been reported to weakly inhibit alpha-glucosidases, the underlying mechanism remains unclear. Therefore, we focused on the regulatory mechanism of dietary glucose absorption: In this study, we aimed to clarify the inhibitory effects of ECH and ACT on sodium-dependent glucose cotransporter (SGLT) 1-mediated gastrointestinal glucose absorption. Uptake experiments were performed using human intestinal Caco-2 cells and the fluorescence glucose analogue, 2-deoxy-2-[(7-nitro-2,1,3benzoxadiazol-4-yDaminc]-n-glucose (2-NBDG). Sodium-dependent 2-NBDG uptake was successfully estimated and this uptake was completely inhibited by an SGLT inhibitor phlorizin. ECH and ACT inhibited sodium-dependent 2-NBDG uptake in a concentration-dependent manner. However, this inhibition was not observed under sodium-free condition. This study suggested that the inhibitory effects of ECH and ACT on SGLT1-mediated glucose uptake contribute to suppression of increased postprandial blood glucose level.
  • Takeo Nakanishi; Yasuhiro Ohno; Rika Aotani; Shio Maruyama; Hiroaki Shimada; Shunsuke Kamo; Hiroko Oshima; Masanobu Oshima; John D. Schuetz; Ikumi Tamai
    SCIENTIFIC REPORTS NATURE PUBLISHING GROUP 7 (1) 16567  2045-2322 2017/11 [Refereed]
    Prostaglandin E-2 (PGE(2)) is associated with proliferation and angiogenesis in colorectal tumours. The role of prostaglandin transporter OATP2A1/SLCO2A1 in colon cancer tumorogenesis is unknown. We evaluated mice of various Slco2a1 genotypes in a murine model of colon cancer, the adenomatous polyposis (APC) mutant (Apc(Delta 716/+)) model. Median lifespan was significantly extended from 19 weeks in Slco2a1(+/+)/Apc(Delta 716/+) mice to 25 weeks in Slco2a1(-/-)/Apc(Delta 716/+) mice. Survival was directly related to a reduction in the number of large polyps in the Slco2a1(-/-)/Apc(Delta 716/+) compared to the Slco2a1(+/+)/Apc(Delta 716/+) or Slco2a1(+/-)/Apc(Delta 716/+) mice. The large polyps from the Slco2a1(-/-)/Apc(Delta 716/+) mice had significant reductions in microvascular density, consistent with the high expression of Slco2a1 in the tumour-associated vascular endothelial cells. Chemical suppression of OATP2A1 function significantly reduced tube formation and wound-healing activity of PGE2 in human vascular endothelial cells (HUVECs) although the amount of extracellular PGE2 was not affected by an OATP2A1 inhibitor. Further an in vivo model of angiogenesis, showed a significant reduction of haemoglobin content (54.2%) in sponges implanted into Slco2a1(-/-), compared to wildtype mice. These studies indicate that OATP2A1 is likely to promote tumorogenesis by PGE2 uptake into the endothelial cells, suggesting that blockade of OATP2A1 is an additional pharmacologic strategy to improve colon cancer outcomes.
  • Masahiro Iwaki; Hiroaki Shimada; Yuri Irino; Manami Take; Sachiko Egashira
    BIOLOGICAL & PHARMACEUTICAL BULLETIN PHARMACEUTICAL SOC JAPAN 40 (6) 926 - 931 0918-6158 2017/06 [Refereed]
    Combination therapy of non-steroidal anti-inflammatory drugs (NSAIDs) and methotrexate (MTX) sometimes triggers adverse effects, such as liver injury, renal failure, gastrointestinal disorders, and myelosuppression, owing to the reduction of MTX clearance. Previous reports have suggested that NSAIDs inhibit renal MTX uptake via organic anion transporters (OATs) and reduced folate transporter (RFC)-1 and efflux via multidrug resistance-associated proteins (MRPs). Recently, our laboratory found inhibitory effects of NSAIDs-glucuronide (NSAIDs-Glu), a major metabolite of NSAIDs, on MRP-mediated MTX transport as a new site of interaction between MTX and NSAIDs. However, it remains unclear that whether NSAIDs-Glu inhibit renal uptake of MTX. Therefore, the present study aimed to evaluate inhibitory effects of several NSAIDs-Glu (diclofenac, R- and S-ibuprofen, R- and S-flurbiprofen, and R- and S-naproxen) on human OAT1 and OAT3-mediated MTX transport. In this study, [H-3]MTX uptake was observed by using human OAT1 and OAT3-overexpressing HEK293 cells in the presence or absence of NSAIDs-Glu. All examined NSAIDs-Glu exhibited concentration-dependent inhibitory effects on MTX uptake via OAT1 and OAT3. Our results indicated that NSAIDs-Glu are more potent (5- to 15-fold) inhibitors of OAT3 than OAT1. Moreover, stereoselective inhibitory effects of NSAIDs-Glu on OATs-mediated MTX uptake were not observed, unlike on MRPs-mediated transport. These findings suggest that inhibition of OAT1 and OAT3-mediated renal uptake of MTX by plasma NSAIDs-Glu may be one of the competitive sites underlying complex drug interaction between MTX and NSAIDs.
  • Atsushi Kawase; Ryota Hashimoto; Mai Shibata; Hiroaki Shimada; Masahiro Iwaki
    INTERNATIONAL JOURNAL OF TOXICOLOGY SAGE PUBLICATIONS INC 36 (3) 260 - 267 1091-5818 2017/05 [Refereed]
    Background and Objectives: Diclofenac (DIC) is metabolized to reactive metabolites such as diclofenac acyl--d-glucuronide (DIC-AG). It is possible that such reactive metabolites could cause tissue damage by formation of covalent protein adducts and other modification of cellular proteins or by induction of immune responses against its covalent protein adducts. However, the detailed mechanisms of idiosyncratic drug-induced liver injury (DILI) have been unclear. The objective is to clarify the involvement of DIC-AG and 4hydroxydiclofenac (4OH-DIC) in acute DILI.Methods:We examined the effects of inhibiting DIC-AG and 4OH-DIC production on covalent protein adduct formation and lactate dehydrogenase leakage using sandwich-cultured rat hepatocytes (SCRHs).Results:After pretreatment of SCRH with (-)-borneol (BOR, a uridine diphosphate (UDP)-glucuronosyltransferase inhibitor) or sulfaphenazole (SUL, a cytochrome P450 2C9 inhibitor) for 30 minutes, intracellular concentrations of DIC, DIC-AG, and 4OH-DIC were determined after further treating cells with 300 M DIC for 3 hours. The decreased levels of reactive metabolites caused by BOR or SUL pretreatment resulted in decreased lactate dehydrogenase leakage from SCRH, although the formation of covalent protein adducts was not affected.Conclusion:These results suggested that both DIC-AG and 4OH-DIC may be involved in acute cytotoxicity by DIC.
  • KAWASE Atsushi; KANETO Ayaka; ISHIBASHI Mao; KOBAYASHI Akihiro; NAKAMURA Hikaru; SHIMADA Hiroaki; IWAKI Masahiro
    Annual Meeting of the Japanese Society of Toxicology The Japanese Society of Toxicology 44.1 O-16  2017 
    【目的】カルボキシ基を含有する薬物による特異体質性肝障害発症の原因として,代謝により生成したアシルグルクロン酸抱合体などの反応性代謝物のタンパク質との共有結合体生成の関与が指摘されている。Diclofenac (DIC)のアシルグルクロン酸抱合体 (DIC-AG)は肝障害の原因となり,その発症に対する免疫系の寄与あるいはglutathione (GSH)との関連は考察されてきたが,これらを組み合わせた状況下でのDIC代謝と毒性発現の関連については不明な点が多い。そこで,本研究では,マウス肝細胞 (Hep)と腹腔マクロファージ (PM)の共培養系を用い,GSH枯渇時の代謝物産生と毒性発現について検討を行った。
    【方法】5週齢雄性ICRマウスよりHepおよびPMを採取し,Hep/PM (1 : 0, 1 : 0.1, または1 : 0.4)を共培養した。GSH枯渇はL-buthionine sulfoximine (BSO)を0.1, 0.3, 1, 3または10 mMで22時間処理させた。DIC代謝の検討では,0.1, 0.3または1 mMでDICを添加した後の培地および細胞内DIC,DIC-AGおよび共有結合体量を測定した。細胞毒性の評価にはLDH漏出量,ATP含量およびカスパーゼ活性を用いた。In vivo試験では,DIC (50 mg/kg, i.p.)単独群,BSO (500 mg/kg, i.p.)併用群,LPS (10 mg/kg)併用群,BSOおよびLPS併用群の血漿および肝臓内DICおよびDIC-AG濃度をHPLC法で測定した。
    【結果および考察】HepにおいてBSO濃度依存的にGSHが枯渇し,GSH枯渇の程度に応じて培地中および細胞内DIC-AG濃度および共有結合体量の有意な上昇がみられた。また,GSH枯渇時にLDH漏出量の有意な上昇が認められ,DICによる毒性はDIC-AGおよび共有結合体量と関連することが示唆された。共培養においてもHep単独と同様にGSH枯渇時に培地中DIC-AG濃度が上昇し,細胞内のATP含量は共培養 (1 : 0.4)のみ有意な低下がみられた。1 : 0.4は炎症時の比率とされ,炎症時にDIC誘発肝毒性が増強する可能性が示された。In vivo試験で血漿中および肝臓内DIC-AG濃度を検討したところ,in vitroと同様にBSOおよびLPS併用群でDIC単独群と比べ顕著に上昇した。以上より,GSH枯渇がDIC-AGの細胞内蓄積を促し,PMにより毒性発現が増強することが示唆された。
  • Taku Kasai; Takeo Nakanishi; Yasuhiro Ohno; Hiroaki Shimada; Yoshinobu Nakamura; Hiroshi Arakawa; Ikumi Tamai
    EXPERIMENTAL CELL RESEARCH ELSEVIER INC 341 (2) 123 - 131 0014-4827 2016/02 [Refereed]
    Chronic inflammation induced by reactive oxygen species is associated with increased risk of developing colorectal cancer (CRC), and prostaglandin E-2 (PGE(2)), which serves as a key mediator of inflammatory responses, plays an important role in CRC initiation and progression. Therefore, in the present study, we aimed to investigate the role of prostaglandin transporter OATP2A1/SLCO2A1 in the changes of PGE(2) disposition in CRC cells in response to oxidative stress. H2O2 induced translocation of cytoplasmic OATP2A1 to plasma membranes in LoVo and COLO 320DM cells, but not in Caco-2 cells. The shift of subcellular OATP2A1 was abolished in the presence of anti-oxidant N-acetyl-L-cysteine or an inhibitor of protein kinase C, which evokes exocytosis. Exposure of LoVo cells to H2O2 caused an increase in the amount of extracellular PGE(2) without changing the sum of intra- and extracellular PGE(2). OATP2A1 knockdown decreased extracellular PGE(2) in LoVo cells. In addition, extracellular PGE(2) was significantly reduced by exocytosis inhibitor cytochalasin D, suggesting that H2O2-induced PGE(2) release occurs in an exocytotic manner. Furthermore, mRNA expression of vascular endothelial growth factor (VEGF) was significantly reduced in LoVo cells by knockdown of OATP2A1. These results suggest that cytoplasmic OATP2A1 likely facilitates PGE(2) loading into suitable intracellular compartment(s) for efficient exocytotic PGE(2) release from CRC cells exposed to oxidative stress. (C) 2016 Elsevier Inc. All rights reserved.
  • Tomoka Gose; Takeo Nakanishi; Shunsuke Kamo; Hiroaki Shimada; Katsumasa Otake; Ikumi Tamai
    PROSTAGLANDINS & OTHER LIPID MEDIATORS ELSEVIER SCIENCE INC 122 10 - 17 1098-8823 2016/01 [Refereed]
    Eicosapentaenoic acid (EPA)-derived prostaglandin E-3 (PGE(3)) possesses an anti-inflammatory effect; however, information for transporters that regulate its peri-cellular concentration is limited. The present study, therefore, aimed to clarify transporters involved in local disposition of PGE(3). PGE(3) uptake was assessed in HEK293 cells transfected with OATP2A1/SLCO2A1, OATP1B1/SLCO1B1, OATP2B1/SLCO2B1, OAT1/SLC22A6, OCT1/SLC22A1 or OCT2/SLC22A2 genes, compared with HEK293 cells transfected with plasmid vector alone (Mock). PGE(3) uptake by OATP2A1-expressing HEK293 cells (HEK/2A1) was the highest and followed by HEK/1B1, while no significantly higher uptake of PGE(3) than Mock cells was detected by other transporters. Saturation kinetics in PGE(3) uptake by HEK/2A1 estimated the K-m as 7.202 +/- 0.595 mu M, which was 22 times higher than that of PGE(2) (K-m = 0.331 +/- 0.131 mu M). Furthermore, tissue disposition of PGE(3) was examined in wild-type (WT) and Slco2a1-deficient (Slco2a1(-/-)) mice after oral administration of EPA ethyl ester (EPA-E) when they underwent intraperitoneal injection of endotoxin (e.g., lipopolysaccharide). PGE(3) concentration was significantly higher in the lung, and tended to increase in the colon, stomach, and kidney of Slco2a1(-/-), compared to WT mice. Ratio of PGE(2) metabolite 15-keto PGE(2) over PGE2 concentration was significantly lower in the lung and colon of Slco2a1(-/-) than that of WT mice, suggesting that PGE(3) metabolism is downregulated in Slco2a1(-/-) mice. In conclusion, PGE3 was found to be a substrate of OATP2A1, and local disposition of PGE(3) could be regulated by OATP2A1 at least in the lung. (C) 2015 Elsevier Inc. All rights reserved.
  • Hiroaki Shimada; Yoshinobu Nakamura; Takeo Nakanishi; Ikumi Tamai
    BIOCHEMICAL PHARMACOLOGY PERGAMON-ELSEVIER SCIENCE LTD 98 (4) 629 - 638 0006-2952 2015/12 [Refereed]
    There is significant evidence that the inducible cyclooxygenase isoform (COX-2) regulates the pericellular concentration of PGE(2); however, the mechanism of the secretory process remains unclear. The present study, therefore, aimed to evaluate the role of prostaglandin transporter (OATP2A1) in PGE(2) secretion from macrophages. lmmunofluorescence staining for Oatp2a1 (Slco2a1) was primarily detected in cytoplasmic domains, and was partially co-localized with anti-PGE(2) antibody, LysoTracker (R), and anti-lysosome-associated membrane protein (Lamp) I antibody in murine macrophage-derived RAW264 cells and peritoneal macrophages (PMs). PGE(2) uptake by subcellular fraction containing light lysosomes was reduced significantly in the presence of an OATP inhibitor and in Slco2a1(+/-) PMs. Secretion of PGE(2) and lysosome-specific N-acetyl-beta-D-glucosaminidase was enhanced in activated macrophagic cells, and diminished significantly under the Ca2+-depleted condition. The amount of PGE(2) secreted from lipopolysaccharide-activated Slco2a1(-/-) PMs was significantly lower than that from PMs from wild type (WT) mice. Expression of Cox-2 and 15-hydroxyprostaglandin dehydrogenase (15-Pgdh) was unchanged between PMs from Slco2a1(-/-) and WT mice. These results suggest that OATP2A1 is involved in PGE(2)-loading into intracellular acidic compartments, including light lysosomes. Thus, OATP2A1 contributes to PGE(2) secretion by macrophages via exocytosis induced by Ca2+ influx, independently of PGE(2) synthesis and metabolism. (C) 2015 Elsevier Inc. All rights reserved.

Conference Activities & Talks

  • ヒト肝ミクロソーム中における アシルグルクロン酸抱合体加水分解特性
    島田 紘明; 生田博之; 阪本健次郎; 中村令奈; 川瀬篤史; 松野純男; 岩城正宏
    日本薬剤学会第37年会  2022/05
  • 肝ミクロソーム中における非ステロイド性抗炎症薬のアシルグルクロン酸 抱合体の加水分解特性
    阪本 健次郎; 島田 紘明; 生田 博之; 川瀬 篤史; 岩城 正宏
  • Ilex latifolia葉抽出物の食後血糖上昇抑制作用
    笹井 剛一; 島田 紘明; 上野 省一; 川瀬 篤史; 森川 敏生; 岩城 正宏
    日本薬学会第142年会  2022/03
  • 横飛 暉斗; 島田 紘明; 吉川 幸伽; 山本 望乃花; 川瀬 篤史; 岩城 正宏
    日本薬学会第142年会  2022/03
  • 島田紘明; 横飛暉斗; 山田爽; 川瀬篤史; 岩城正宏
    第71回 日本薬学会関西支部総会・大会  2021/10
  • 非ステロイド性抗炎症薬のアシルグルクロン酸抱合体のラットおよびヒト肝ミクロソーム中における加水分解
    生田博之; 島田紘明; 川瀬篤史; 岩城正宏
    第71回 日本薬学会関西支部総会・大会  2021/10
  • Ilex latiforiaによる食後血糖上昇抑制作用
    島田紘明; 笹井剛一; 井芹太一; 森川敏生; 上野省一; 岩城正宏
    第21回 日本抗加齢医学会  2021/06
  • 横飛暉斗; 島田紘明; 吉川幸加; 川瀬篤史; 岩城正宏
    第 63 回 日本脂質生化学会  2021/06
  • Ilex latiforiaによる食後血糖上昇抑制作用
    島田 紘明; 井芹 太一; 笹井 剛一; 上野 省一; 岩城 正宏
    日本薬学会第140年会  2020/03
  • 非ステロイド性抗炎症薬のアシルグルクロン酸抱合体生成に対するCa2+およびMg2+の影響  [Not invited]
    第68回 日本薬学会近畿支部総会・大会  2018/10
  • '次世代'の薬剤学教員が考える薬学教育  [Invited]
    日本薬剤学会第33年会  2018/06
  • 15-PGDHおよびOATP2A1の発現変動は肝障害時の肝組織中PGE2量を調節する  [Not invited]
    島田紘明; 橋本凌太; 青木彩; 東田千代; 川瀬篤史; 岩城正宏
    日本薬物動態学会 第32回年会  2017/11
  • 砂漠人参カンカニクジュヨウの血糖上昇抑制作用メカニズムの解明  [Not invited]
    島田紘明; 卜部裕一; 岡本雄平; 李征; 川瀬篤史; 森川敏夫; 村岡修; 岩城正宏
    第39回生体膜と薬物の相互作用シンポジウム  2017/10
  • カンカニクジュヨウ中主成分エキナコシド,アクテオシドのグルコース/Na+共輸送トランスポーター阻害作用  [Not invited]
    島田紘明; 卜部裕一; 岡本雄平; 川瀬篤史; 李征; 森川敏生; 村岡修; 岩城正宏
    日本生薬学会第64回年会  2017/09
  • 薬物誘発性肝障害に伴う肝プロスタグランジンE2動態変動  [Not invited]
    橋本凌太; 島田紘明; 東田千代; 青木彩; 川瀬篤史; 岩城正宏
    第59回 日本脂質生化学会  2017/06
  • The Role of OATP2A1/SLCO2A1 in PGE2 Secretion from Macrophages.  [Invited]
    Hiroaki Shimada; Takeo Nakanishi; Yoshinobu Nakamura; Shio Maruyama; Ikumi Tamai
    日本薬物動態学会 第30回年会  2015/11
  • Role of OATP2A1/SLCO2A1 in PGE2 secretion from macrophages  [Not invited]
    Hiroaki Shimada; Yoshinobu Nakamura; Takeo Nakanishi; Ikumi Tamai
    2015金沢大学・釜山大学校・瀋陽薬科大学 薬学系国際合同シンポジウム  2015/10
  • マクロファージからのPGE2開口分泌におけるプロスタグランジン輸送体(PGT)の役割  [Not invited]
    島田紘明; 中西猛夫; 中村吉伸; 玉井郁巳
    第56回 日本脂質生化学会  2014/06


Industrial Property Rights

  • 特許7015463:グルコース輸送阻害剤及びグルコース輸送阻害用の機能性食品    2022/02/15
    岩城正宏, 島田紘明, 村岡修, 森川敏生, 二宮清文
  • 特開2021-138647:血糖値上昇抑制剤  2021/09/16
    上野 省一, 岩城 正宏, 島田紘明

Awards & Honors

  • 2020/10 日本薬学会関西支部 優秀ポスター賞
     Acetaminophen誘発性肝障害における肝Prostaglandin E2量調節機構とその意義 
    受賞者: 横飛 暉斗;島田 紘明;吉川 幸加;川瀬 篤史;岩城 正宏
  • 2016/10 日本薬物動態学会 第31回年会 ベストポスター賞
     マウス胎盤内プロスタグランジン E 2 分解に果たす輸送体の役割 
    受賞者: 稲垣舞;西村友宏;中西猛夫;島田紘明;赤沼伸乙;立川正憲;細谷健一;玉井郁巳;中島恵美;登美斉俊
  • 2016/10 第10回日本薬局学会学術総会 ポスター優秀演題賞
    受賞者: 松浦正佳;岸本理咲;大島徹;加治工衛;前多智昭;川瀬篤史;島田紘明;岩城正宏

Research Grants & Projects

Teaching Experience

  • Elementary Chemical EnglishElementary Chemical English Kindai University


  • 2017/04 -2017/04  カンカニクジュヨウ中成分エキナコシド、アクテオシドの食後血糖上昇抑制メカニズムの解明 
    平成29年度 近畿大学学内研究助成金 SR18

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