Oltipraz, a synthetic dithiolethione, has chemopreventive effect through nuclear factor erythroid 2-related factor 2 (Nrf2) activation. Nrf2 is known to be involved in the development of experimental steatohepatitis in rodents. In this study, to evaluate the effect of oltipraz on lipid and bile acid metabolism, wild-type and Nrf2-null mice were fed the standard diet (containing 4% soybean oil) with or without oltipraz. Based on these results, we examined the effect of oltipraz on the experimental steatohepatitis in high-fat diet (containing 4% soybean oil and 20% lard) fed Nrf2-null mice. Oltipraz induced hepatic mRNA expression of peroxisome proliferator-activated receptor α, carnitine palmityl transferase 1, and bile salt export pump by Nrf2 independent mechanisms. In Nrf2-null mice fed a high-fat diet for 12 weeks, moderate to severe inflammation and fibrosis were observed. Oral administration of oltipraz suppressed the degree of inflammation and fibrosis in Nrf2-null mouse liver fed a high-fat diet. These histopathological findings approximately corresponded to the data of mRNA expression of tumor necrosis factor α, monocyte chemoattractant protein-1, Timp-1, and collagen type 1α1. These results indicated that oltipraz administration ameliorated liver injury by Nrf2 independent manner in a model of steatohepatitis generated by Nrf2-null mice with high-fat diet.

Ezetimibe and pemafibrate are lipid-lowering drugs and promote reverse cholesterol transport. However, it is unknown whether cholesterol is mainly excreted by hepatobiliary excretion or by non-biliary transintestinal cholesterol efflux (TICE). We evaluated the effects of ezetimibe and pemafibrate on hepatic and intestinal cholesterol transporter regulation in Sham-operated rats, and examined the effects of these drugs on TICE in bile duct-ligated rats. Seven-week-old male Sprague-Dawley rats were treated as follows for two weeks: 1) Sham, Sham operation; 2) BDL, bile duct ligation; 3) E-Sham, Sham + ezetimibe; 4) E-BDL, BDL + ezetimibe; 5) P-Sham, Sham + pemafibrate; and 6) P-BDL, BDL + pemafibrate. Blood, liver, jejunum, and feces were collected 72 h post-surgery. Hepatic cholesterol levels were decreased in P-Sham and E-Sham, and were lower in E-BDL and P-BDL than in BDL. Fecal cholesterol levels increased in E-Sham and P-Sham compared with Sham, and were higher in E-BDL and P-BDL than in BDL. In liver, Abcg5 mRNA showed induction in E-Sham, Abcg5 and Abca1 mRNA were induced in P-Sham, Abcg5 mRNA was reduced in E-BDL, and Abca1 mRNA was increased in P-BDL. In jejunum, Abcg5 mRNA was induced in E-Sham. Abcg8 mRNA was induced in E-Sham and P-Sham. NPC1L1 mRNA showed reduced expression in P-Sham and P-BDL. SR-B1 mRNA was reduced in P-Sham, and the expression decreased in P-BDL. LDL receptor mRNA was induced in BDL and P-BDL. Ezetimibe and pemafibrate may promote TICE by increasing Abcg5/g8, while pemafibrate may inhibit intestinal cholesterol absorption by decreasing SR-B1 and NPC1L1.

BACKGROUND: The automated hematology analyzer Celltac G (Nihon Kohden) was designed to improve leukocyte differential performance. Comparison with analyzers using different leukocyte detection principles and differential leukocyte count on wedge film (Wedge-Diff) shows its clinical utility, and comparison with immunophenotypic leukocyte differential reference method (FCM-Ref) shows its accuracy performance. METHODS: For method comparison, 598 clinical samples and 46 healthy volunteer samples were selected. The two comparative hematology analyzers (CAAs) used were XN-9000 (Sysmex) and CELL-DYN Sapphire (Abbott). The FCM-Ref provided by the Japanese Society for Laboratory Hematology was selected, and a flow cytometer Navios (Beckman-Coulter) was used. In manual differential, two kinds of automated slide makers were used: SP-10 (Sysmex) for wedge technique and SPINNER-2000 (Lion-Power) for spinner technique. The spinner technique avoids the issue of Wedge-Diff smudge cells by removing the risk of breaking cells and non-uniformity of blood cell distribution on films (Spinner-Diff). RESULTS: The Celltac G showed sufficient comparability (r = 0.67-1.00) with the CAAs for each leukocyte differential counting value at 0.00-40.87(109 /L), and sufficient comparability (r = 0.73-0.97) with FCM-Ref for each leukocyte differential percentage at 0.4-78.5. The identification ratio of the FCM-Ref in CD45-positive cells was 99.7% (99.4% to 99.8%). Differences were found between FCM-Ref/Celltac G/XN-9000/Spinner-Diff and Wedge-Diff for monocytes and neutrophils. The appearance ratio of smudge cells on wedge and spinner film was 12.5% and 0.5%. CONCLUSION: The Celltac G hematology analyzer's leukocyte differential showed adequate accuracy compared with the CAAs, FCM-Ref, and two manual methods and was considered suitable for clinical use.

INTRODUCTION: The hematology analyzer, Celltac G (Nihon Kohden), designed to improve platelet count (Plt) accuracy, is equipped with new sheath flow control technology. Clinical evaluation of the Celltac G was assessed by comparability with XN-9000 (Sysmex Corporation) and CELL-DYN Sapphire (Abbott Diagnostics). The accuracy of all three analyzers, which use different measuring principles, was compared with the immunoplatelet reference method (FCM-Ref). METHODS: Repeatability and within-laboratory imprecision were assessed using 10 clinical fresh whole blood samples and three control materials with differing levels. Carryover was evaluated using 6 clinical fresh whole blood samples. For method comparison between the three analyzers, 388 samples were used. Plt accuracy among the three analyzers was evaluated using 54 blood samples, including 42 samples with a platelet count less than 50x109 /L. The International Council for Standardization in Haematology method for Plt was used as the FCM-Ref. RESULTS: The Celltac G showed sufficient performance with regard to imprecision, carryover, and comparability. The Analytical Measurement Interval (AMI) and linearity for all parameters of Plt were validated within 4.6 to 809.1 (×109 /L). All hematology analyzers showed some disagreement in Plt when compared with the immunoplatelet reference method. CONCLUSION: The Celltac G hematology analyzer is suitable for clinical use. Platelet count evaluation of the three analyzers suggests the need to determine a reportable measurement interval (RMI) in the clinical laboratory for adequate reporting of a Plt from multiple different values.

Besides diet therapy, hypolipidemic pharmacological therapy may be a crucial component of nonalcoholic fatty liver disease (NAFLD) treatment. Ezetimibe may be a promising drug for treatment of NAFLD. N-3 polyunsaturated fatty acids, which are abundant in fish oil, reduces serum and hepatic cholesterol and triglycerides in rodents. The aim of this study was to examine the combined effects of dietary fish oil and ezetimibe on lipid metabolism in rats. Seven-week old male SD rats were allocated to four different diets containing 1) 10% soybean oil (C), 2) 10% fish oil (F), 3) 10% soybean oil + 0.005% ezetimibe (E), and 4) 10% fish oil + 0.005% ezetimibe (F+E) for 4 weeks, when the liver, jejunum, blood, and fecal samples were collected. Compared to the C group, the F+E-diet decreased hepatic triglycerides and cholesterol 84% and 86%, but it did not increase fecal cholesterol. In liver, the expression of lipogenic enzymes were decreased in F+E diet, whereas β-oxidation related genes were not increased. Abcg5/g8 mRNA expression were increased 1380%/442% when ezetimibe was added to the F-diet. These gene expression changes are related to the decrease in hepatic lipids. In jejunum, Abcg5/g8 mRNA were increased 244%/841% when ezetimibe was added to the F-diet. Hepatic induction of Abcg5/8 rather than intestinal induction correlates with the marked decrease in liver cholesterol when ezetimibe was added to the F-diet. These data suggest that fish oil diet and ezetimibe in combination may be beneficial therapy for NAFLD by increasing hepatic Abcg5/g8 and decreasing lipogenic genes. SIGNIFICANCE STATEMENT: There is currently no single treatment for NAFLD. Thus, lifestyle modifications including dietary regulation and physical activity are also important options. In this study, ezetimibe, a cholesterol absorption inhibitor, was evaluated for the treatment of liver steatosis in rats fed on the different diets. We found that ezetimibe and fish oil in combination markedly improved fatty liver by increasing cholesterol efflux transporters. The combination therapy of fish oil agents and ezetimibe may be effective for NAFLD.

Streptococcus pneumoniae is one of the most common bacteria causing community-acquired pneumonia and meningitis. The use of 7-valent pneumococcal conjugate vaccine (PCV7) has reduced the incidence of pneumococcal disease while changing pneumococcal population through herd immunity and non-vaccine pneumococci replacement. This study investigated molecular epidemiologic characteristics of pneumococcal strains in the Kinki region of Japan from 2008 to 2013. A total of 159 invasive pneumococcal isolates were characterized by serotyping, antibiotic susceptibility testing, PCR analysis of penicillin-binding protein genes, multilocus sequence typing (MLST), and pulsed-field gel electrophoresis (PFGE). In adult populations, pediatric PCV7 introduction decreased isolates expressing PCV7 serotypes via herd immunity and increased isolates expressing non-PCV7 serotypes. The rate of penicillin resistance and isolates with alterations in all three pbp genes was higher in PCV7 type isolates than in non-PCV7 type isolates. In MLST analysis, all of serotype 19F isolates were of the same sequence type, ST236, which is the antimicrobial-resistant clone Taiwan19F-14, and the majority of serotypes 23F and 19A isolates were of ST1437 and ST3111 respectively, which are the predominant clones of antimicrobial-resistant pneumococci in Japan. In PFGE profiles, serotype 6B-ST2224, serotype 19F-ST236, serotype 19A-ST3111, and serotype 23F-ST1437 formed six separate clusters composed of genetically identical strains, and genetically identical serotype 22F-ST433 formed two different clusters between the pre- and post-PCV7 period. The results of molecular analysis suggest the spread and persistence of these identical antimicrobial resistant clones in the Kinki region and genetic changes of epidemic clone serotype 22F-ST433 before and after pediatric PCV7 introduction.

70代男性,左大腿基底細胞癌術後にジフテリア毒素産生性Corynebacterium ulceransによる手術部位感染を発症した症例を報告する。患者はジフテリア様症状を呈することはなく,創部洗浄のみで軽快した。室内飼育のネコの鼻腔と咽頭からもC.ulceransが分離され,MLSTにより患者由来株と同じくST2に分類されたため,飼いネコと患者間で水平伝播したと考えられた。Corynebacterium spp.は常在菌として扱われるため菌種レベルの同定がされない場合が多いが,β-溶血を示す集落は本菌を考慮し,菌種レベルの同定を行う必要がある。(著者抄録)

Maternal high-fat diet (HFD) consumption during pregnancy and lactation affects metabolic outcomes and lipid metabolism of offspring in later life in a gender-specific manner. However, it is not known whether maternal HFD alters bile acid metabolism in adult mice offspring. The purpose of this study was to elucidate the relationship between maternal HFD-induced metabolic diseases and bile acid metabolism in male and female adult mice offspring. Female mice were fed either standard chow (C) or HFD (H) for 10 weeks pre-pregnancy until lactation. After weaning, offspring were fed a chow diet until 11 weeks of age, then challenged with either C or H diet for 4 weeks, and divided into eight groups in accordance with mother’s and offspring’s diets: male(M) CC, MHC, MCH, MHH, female(F) CC, FHC, FCH, and FHH. MHH showed greater weight gain compared to FHH. Liver weight was higher in MHH than in FHH. Serum total cholesterol levels were higher in MHH than in MHC, and tended to be higher in MHH than in FHH. Serum glucose levels were higher in MHH than in MHC. Hepatic triglyceride levels were higher in MHH than in MHC. Hepatic mRNA expression of bile acid uptake transporters Oatp1a1 and Oatp1b2 was increased in MHH, compared to MCH. Hepatic mRNA expression of HMGCoAR, Cyp7a1, Sult2a1, and Oatp1a4 was increased in FHH, compared to FCH. In conclusion, maternal HFD consumption may promote bile acid synthesis, sulfation and excretion in female offspring fed a HFD, which may confer resistance to HFD-induced metabolic phenotypes.

The 7-valent pneumococcal conjugate vaccine (PCV7) was introduced among children in Japan in 2010. There are no long-term multicenter surveillance studies of antimicrobial resistance in S. pneumoniae before and after the introduction of PCV7. Therefore, we examined chronological trends in antimicrobial resistance among 4534 strains of S. pneumoniae isolated from both children and adults in the Kinki region of Japan during 2001–2015. High-level penicillin and third-generation cephalosporin resistance in S. pneumoniae increased among both children and adults during the period before the introduction of PCV7 (2001–2010). Besides penicillin and cephalosporin, pneumococcal carbapenem and macrolide resistance increased among children. The rate of resistance to these antibiotics was higher among children than among adults. The introduction of PCV7 decreased the rate of non-susceptibility to β-lactam antibiotics and the rate of multidrug resistant S. pneumoniae among children, but not among adults.

高フルクトース食摂取ラットにおいて油脂源が大豆油の場合よりオリーブ油の場合の方が、血中トリグリセリド及び遊離脂肪酸濃度の上昇がもたらされ肝臓脂質が上昇することを報告した。

Background: We encountered a rare case of Waldenstrom macroglobulinemia with temporary appearance of 7S IgM half molecule and with monoclonal proteins binding to agarose gel. Methods: The patient's serum and urine were analyzed using sodium dodecyl sulfate-polyacrylamide gel electrophoresis and immunoblotting. The N-terminal amino acid sequences of the IgM with abnormal mass (68 kDa) were determined and compared with those of known immunoglobulin. Results: The 68 kDa IgM consisted of a defective Et chain (36 kDa) and an intact kappa chain. N-terminal amino acid sequence analysis demonstrated that the defective It chain had the variable region of IgM. The agarose gel-binding ability of the IgM-w M-protein was lost after reduction or alkaline treatment of serum. Conclusions: The 7S half molecule IgM in the present case may miss a large part of the constant region of the mu chain.

Objective The menstrual cycle-related changes in clinical laboratory values were analysed by use of data obtained in the Asian multicentre study aimed at derivation of common reference intervals for 85 major clinical laboratory tests. Methods Among 1876 healthy female volunteers, 893 had regular menstruation. They were classified into five groups according to dates between sample collection and the start of the last menstrual cycle: early follicular phase (1-6 days), late follicular phase (7-12 days), ovulatory phase (13-16 days), early luteal phase (17-22 days), and late luteal phase (23-31 days). Multiple linear regression analysis was performed to evaluate the menstrual cycle-related changes in test results. The magnitude was expressed as a standard deviation ratio of between-phase standard deviation to between-individual standard deviation based on nested ANOVA. Results Aside from obvious changes for four sex hormones (oestradiol, progesterone, follicle-stimulating hormone, and luteinizing hormone), we observed statistically significant menstrual cycle-related changes in the following tests (standard deviation ratio >0.15): Na, Cl, creatine kinase, C-reactive protein, serum amyloid A, carbohydrate antigen 125, and parathyroid hormone were higher during the early follicular phase, while insulin, total cholesterol, and white blood cell were higher during the luteal phase. Significant associations of those test items with the four sex hormones were revealed. Conclusions The menstrual cycle-related changes in laboratory test results were revealed in some commonly tested items other than sex hormones. The findings are of interest in understanding female physiology in relation to hormonal changes, but the magnitude of changes is rather small and not very relevant in interpreting test results.

Geranium dielsianum (GD) is a perennial plant found in the Andean highlands of Peru. Over the decades, the decoction of GD has been consumed as a tea protective against diabetes. Furthermore, GD has traditionally been used as an anti-diabetic, anti-inflammatory, and anti-diarrheal folk medicine. However, there is little scientific evidence elucidating its effects. This study aimed to test the effect of the GD extract on the intestinal environment. In male Sprague-Dawley rats, GD extract increased levels of Bifidobacteria and Lactobacilli and decreased levels of Clostridium leptum subgroup and Bacteroides group in the intestine. Furthermore, 3-hydroxyphenylacetic acid was present at high concentrations in the caecal content of GD extract intake group. Through the above changes in microbiota and microbial metabolites, faecal bulk and moisture increased and caecal pH and putrefactive products decreased. These results suggest that GD extract has a potent prebiotic effect. (C) 2014 The Authors. Published by Elsevier Ltd.

Nuclear factor-E2-related factor 2 (Nrf2) is a regulator of lipid metabolism as well as various cytoprotective enzymes and may be involved in the pathogenesis of non-alcoholic fatty liver disease. Although, bile acids affect lipid metabolism, the role of Nrf2 in bile acid metabolism remains unclear. In this study, it was tested how Nrf2 modulates lipid and bile acid homeostasis in liver in response to changes of cholesterol absorption under high-fat diet using Nrf2-null mice. Eight-week-old male wild-type and Nrf2-null mice (n = 6/group) were divided into three groups fed the following diets: 1) control diet containing 4% soybean oil and 16% lard, 2) control diet plus ezetimibe, 3) control diet plus cholesterol. Blood and livers were removed after 4 weeks feeding. High cholesterol diet increased hepatic expression of liver X receptor a target genes related to fatty acid metabolism (FAS, ACC1, SREBP-1c, SCD-lc and CD36), cholesterol transport (Abcg5/abcg8) and bile acid synthesis (Cyp7a1) in wild type mice. However, these genes were not induced in Nrf2-null mice. These findings suggest that Nrf2 has a relation to liver X receptor a and controls the regulation of gene expressions related to lipid and bile acid metabolism.

Bence Jonesタンパク(BJP)は尿タンパク試験紙(以下、試験紙)にはほとんど反応しないことが定説となっている。我々はこの説に疑問を持ち、過去約6年間におけるBJP定性試験(Putnam法)陽性で、免疫電気泳動法(IEP)あるいは免疫固定法(IFE)においてBJP陽性が確認されている患者尿の試験紙反応性について調査した結果、BJP含有尿352検体(66症例)中、試験紙陰性はわずかに10検体(2.8%)で、342検体(97.2%)が±以上の反応(±〜4+)であった。試験紙法陰性検体は経過観察中の多発性骨髄腫患者3例のみであった。しかも、尿総タンパク量は、15mg/dL未満であり、試験紙法の検出限界濃度以下であった。またIgGおよびアルブミンが出現していないBJP陽性尿を用い、試験紙が±以上に反応する濃度(検出限界)を求めた結果、アルブミンの検出感度と同等で15mg/dLであった。さらにIgGが試験紙に反応しがたいことを再確認したうえで、IgGを還元処理してL鎖を遊離させた後は、試験紙の反応性が強くなることを確認した。したがって、尿タンパク試験紙がBJPに反応しない(反応しがたい)という説は見直されるべきであると考える。(著者抄録)

Accumulating evidence suggests that ezetimibe may be a promising agent for treatment of nonalcoholic fatty liver disease and steatohepatitis (NAFLD/NASH). Phlebotomy and dietary iron restriction reduce serum transaminase in NAFLD/NASH patients. Recent studies have shown that a mutual effect exists between lipid metabolism and iron metabolism. Accordingly, we examined the effect of ezetimibe on iron metabolism in mice fed a high-fat diet with or without iron. We fed C57BL/6 mice the following diets for 12 weeks. Experiment 1 comprised [1] a control diet (C), [2] C plus ezetimibe (0.3 mg/day; 4 weeks) (CE), [3] a high-fat diet (H), and [4] H plus ezetimibe (HE). Experiment 2 comprised [1] C containing carbonyl iron (average; 22.4 mg/day; 6 weeks) (CI), [2] CI plus ezetimibe (CIE), [3] H containing carbonyl iron (HI), and [4] HI plus ezetimibe (HIE). Blood, livers, and duodenum were removed after 12 weeks. In experiment 1, the hepatic iron levels were higher in HE than H, whereas there was no difference between C and CE. Hepatic mRNA expression of transferrin receptor 1 and 2, ferritins, and hepcidin were increased more in CE than C, and more in HE than H. In the duodenum, divalent metal transporter 1, ferritin H, and hephaestin mRNA levels were increased in CE compared with C. In experiment 2, hepatic iron concentrations were higher in HIE than HI. Hepatic mRNA expression of ferritin L and hepcidin were increased in HIE compared with HI. In duodenum, ferritin L mRNA was increased in HIE compared with CIE. Ezetimibe induced hepatic iron uptake transporter expression in mice fed a high-fat diet, causing increased hepatic iron concentrations.

脂質異常症に関する基礎・臨床研究の最新知見として、脂質関連マーカー測定と薗臨床的意義について、リポタンパク分画精密測定を中心として解説した。

Background: Bilirubin has antioxidant properties and is known to have a role in the prevention of cardiovascular disease. Bilirubin is known to be negatively associated with serum parameters related to atherosclerosis. Triglyceriderich lipoprotein is known to play an important role in the progression of atherosclerosis. The aim of the present study was to elucidate the relationship between serum bilirubin and serum remnant lipoprotein cholesterol (RLP-c). Methods: We examined 270 Japanese males (mean age 64.8 years) without liver dysfunction and anemia from the Outpatient Clinic of Kinki University Hospital. Serum bilirubin, cholesterol, LDL cholesterol, HDL cholesterol, triglyceride and RLP-c levels were measured. Results: Serum RIP-c level was significantly lower in patients in the high bilirubin group than those in the low bilirubin group. Conclusions: The findings of this study show that serum bilirubin within physiological levels may affect the serum RLP-c and suggest bilirubin may have a role in preventing atherosclerosis.

A pseudo-outbreak of Mycobacterium lentiflavum occurred that contaminated the tap water at the sputum collection booth in our hospital. M. lentiflavum was detected from 81 patients, but the infectious patients were not recognized. M. lentiflavum was cultured from 6 of 103 hospital tap water samples excluding the sputum collection booth. Twenty two isolates (14 clinical isolates, 3 isolates from the sputum collection booth, and 5 isolates from hospital tap water) were classified into profiles A–E by automated repetitive sequence-based PCR. The clinical isolates and the environment isolates included both profiles C and D. The similarity percentages were more than 95%.

Background and Aim: Hepatic excessive iron may play a role in the pathogenesis of non-alcoholic steatohepatitis (NASH). Nrf2 is a master regulator of antioxidative responses. However, the role of Nrf2 in lipid and iron homeostasis remains unclear. Accordingly, it was examined how Nrf2 regulates lipid-related and iron-regulatory genes after feeding a high-fat diet (HFD) with iron. Methods: Wild-type and Nrf2-null mice were fed the following diets: (i) control diet (4% soybean oil) for 12 weeks, (ii) control diet for 8 weeks followed by control diet containing 0.5% carbonyl iron for 4 weeks, (iii) HFD (4% soybean oil and 16% lard) for 12 weeks, (iv) HFD for 8 weeks followed by HFD containing 0.5% carbonyl iron for 4 weeks. Blood and livers were removed after 12 weeks. Results: Nrf2-null control mice exhibited a tendency towards higher hepatic triglycerides compared to wild-type control mice. Hepatic malondialdehyde was higher and hepatic iron levels tended to be higher in Nrf2-null mice than wild-type counterparts while on a HFD. The HFD with iron synergistically induced mRNA expression of Ppara targets, including Acox and Cpt1 in wild-type mice, yet the induction was diminished in Nrf2-null mice. Hepatic hepcidin and ferroportin 1 mRNA expression were increased in wild-type mice after feeding a HFD with iron, but were unchanged in any group of Nrf2-null mice. Conclusions: Nrf2 deletion dysregulates hepatic mRNA expression of beta-oxidation enzymes and iron-related genes, possibly causing a trend for increased hepatic triglyceride and iron concentrations. Nrf2 may have roles in the progression of NASH.

Aim: Fish oil rich in n-3 polyunsaturated fatty acids is known to affect hepatic lipid metabolism. Several studies have demonstrated that fish oil may affect the bile acid metabolism as well as lipid metabolism, whereas only scarce data are available. The aim of this study was to investigate the effect of fish oil on the gene expression of the transporters and enzymes related to bile acid as well as lipid metabolism in the liver and small intestine. Methods: Seven-week old male C57BL/6 mice were fed diets enriched in 10% soybean oil or 10% fish oil for 4 weeks. After 4 weeks, blood, liver and small intestine were obtained. Results: Hepatic mRNA expression of lipids (Abcg5/8, multidrug resistance gene product 2) and bile acids transporters (bile salt export pump, multidrug resistance associated protein 2 and 3, organic solute transporter a) was induced in fish oil-fed mice. Hepatic Cyp8b1, Cyp27a1 and bile acid CoA : amino acid N-acyltransferase were increased in fish oil-fed mice compared with soybean-oil fed mice. Besides, intestinal cholesterol (Abcg5/8) and bile acid transporters (multidrug resistance associated protein 2 and organic solute transporter a) were induced in fish oil-fed mice. Conclusion: Fish oil induced the expression of cholesterol and bile acid transporters not only in liver but in intestine. The upregulation of Abcg5/g8 by fish oil is caused by an increase in cellular 27-HOC through Cyp27a1 induction. The hepatic induction of bile acid synthesis through Cyp27a1 may upregulate expression of bile acid transporters in both organs.

We evaluated the diagnostic utility of peripheral blood neutrophil distribution patterns obtained using the CELL-DYN SAPPHIRE hematology analyzer in patients with myelodysplastic syndrome (MDS). Peripheral blood was obtained from 467 individuals including 32 patients with MDS, and the respective neutrophil distribution patterns were observed using two light scatters [7-degree complexity (7D) and 90-degree lobularity (90D)]. These scattering intensities are shown as median (median neutrophil distribution: MND) and coefficient of variation (neutrophil distribution width: NDW). Generally, MDS patients showed lower 7D MND, higher 7D NDW, lower 90D MND and higher 90D NDW than other comparable groups. Whereas 90D parameters were more diagnostically efficient than 7D ones in patients with MDS. The sensitivity and specificity of 90D MND for MDS patients became 78.1 and 78.9%, respectively (cut-off value = 14,514). 90D NDW was most diagnostically effective with 87.5% sensitivity and 91.0% specificity (cut-off value = 21.2%). Both 90D parameters showed no evident correlation with the degree of either leukocytopenia or peripheral blood dysgranulopoiesis. In conclusion, neutrophil distribution parameters, especially 90D NDW, appear to provide convenient and objective markers for the screening of patients with MDS in routine laboratory hematology.

As interprofessional work in cancer treatment becomes increasingly important, medical technologists are required to play active roles as part of the team. The cancer center of our hospital organized a lecture meeting, "The 6th Lecture Meeting: Living Together," for cancer patients and their families, and a medical technologist presented a lecture entitled: "Cancer treatment and clinical examinations." According to the results of a questionnaire survey conducted following the meeting, most participants were able to understand the lecture and were satisfied with it. Based on the opinions expressed by meeting participants and the questionnaire results, medical technologists initiate the following services and activities: Circled digit one They explain the results of white blood cell and neutrophil counts of patients on chemotherapy and/or radiation therapy and Circled digit two provide medical examinations and consultation at outpatient chemotherapy centers. We believe that these efforts will help improve cancer treatment and further contribute to interprofessional health care.

The innate immunity to viral infections induces a potent antiviral response mediated by interferons (IFN). Although IFN-gamma is detected during the acute stages of illness in the upper respiratory tract secretions and in the serum of influenza A virus-infected individuals, control of influenza A virus is not dependent upon IFN-gamma as evidenced by studies using anti-IFN-gamma Ab and IFN-gamma(-/-) mice. Thus, we hypothesized that IFN-gamma is not critical in host survival because influenza A virus has mechanisms to evade the antiviral activity of IFN-gamma. To test this, A549 cells, an epithelial cell line derived from lung adenocarcinoma, were infected with influenza virus strain A/Aichi/2/68 (H3N2) (Aichi) and/or stimulated with IFN-gamma to detect IFN-gamma-stimulated MHC class II expression. Influenza A virus infection inhibited IFN-gamma-induced up-regulation of HLA-DR alpha mRNA and the IFN-gamma induction of class II transactivator (CIITA), an obligate mediator of MHC class II expression. Nuclear translocation of Stat1 alpha upon IFN-gamma stimulation was significantly inhibited in influenza A virus-infected cells and this was associated with a decrease in Tyr701 and Ser727 phosphorylation of Stat1 alpha. Thus, influenza A virus subverts antiviral host defense mediated by IFN-gamma through effects on the intracellular signaling pathways.

Background and Aim: To evaluate the importance of cholesterol and bile acid concentrations in the intestinal lumen to cholesterol homeostasis, we investigated the effect of cholesterol-, bile salt- or cholestyramine-administration on the regulation of intestinal mRNA related to cholesterol and bile acid metabolism. Methods: Male Wistar rats fed on standard laboratory chow (AIN-93) were allocated into four experimental groups: (i) control group; (ii) cholesterol group (CH), which was fed cholesterol in diet (2% [w/w]) for 14 days; (iii) cholic acid (CA) group, which was fed CA in diet (1% [w/w]) for 14 days; (iv) cholestyramine (CT) group, which was fed CT in diet (5% [w/w]) for 14 days. Blood, liver and small intestine were obtained after 14 days. Serum lipids and bile acids were measured by colorimetric assays, and hepatic and intestinal mRNA relating to lipid and bile acid metabolism was studied by reverse transcription-polymerase chain reaction. Results: Intestinal ABCG8, liver X receptor alpha, small heterodimer partner (SHP) and sterol regulatory element binding protein 1c (SREBP-1c) mRNA expressions were markedly increased in the CH group. Intestinal multidrug resistance associated protein (MRP) 2 and MRP3 mRNA expressions were markedly increased in the CA group. In the CT group intestinal MRP2, ABCG5, ABCG8, SHP and SREBP-1c mRNA expressions were markedly decreased. Conclusion: The bile acid availability in the intestinal lumen alters the expression of various intestinal mRNA relating to not only bile acid metabolism, but also lipid metabolism. Though the mechanism of the intestinal SHP elevation is unclear, cholesterol feeding may affect the intestinal bile acid metabolism via intestinal SHP expression.

Osthol, a coumarin compound, was isolated from the dried fruits of Cnidium monnieri (Umbelliferae) and the effect of dietary osthol on hypertension and lipid metabolism was examined in stroke-prone spontaneously hypertensive rats (SHRSP). Six-week-old male SHRSP were fed the experimental diet containing 0.05% osthol by weight for 4 weeks with free access to the diet and water. Elevation of systolic blood pressure was significantly suppressed on and after 3 weeks. In addition, significant decreases in cholesterol and triglyceride contents in the liver were recognized without any significant changes in serum lipids profiles. A comparative study on hepatic mRNA expression indicated that osthol induced a significant increase in 3-hydroxy-3-methylglutaryl coenzymeA (HMG-CoA) reductase mRNA expression, which may lead to decrease in hepatic cholesterol pool through inhibition of the enzyme activity. Moreover, osthol induced a significant increase in acyl-CoA oxidase mRNA expression associated with an increase in carnitine palmitoyl transferase I a mRNA expression, which suggests the acceleration of P-oxidation of hepatic fatty acids. This may be responsible, at least in part, for the reduction of hepatic triglyceride content in SHRSP. These beneficial effects of osthol could be useful for both prevention of atherosclerosis and suppression of hepatic lipid accumulation. (C) 2007 Elsevier Ireland Ltd. All rights reserved.

1. Recently, we reported that 4-hydroxyderricin, one of the major chalcones in Angelica keiskei extract (ethyl acetate extract from the yellow liquid of stems), exerted hypotensive and lipid regulatory actions in stroke-prone spontaneously hypertensive rats (SHRSP). In the present study, we isolated xanthoangelol, another major chalcone in A. keiskei extract, and examined the effect of dietary xanthoangelol on blood pressure and lipid metabolism in SHRSP. 2. Six-week-old male SHRSP were fed diets containing 0.02% or 0.1% xanthoangelol (0.02 and 0.10 Xan, respectively) for 7 weeks, with free access to the diet and water. There were no significant changes in daily food intake, bodyweight or systolic blood pressure throughout the experimental period. Serum total cholesterol levels tended to decrease in the two experimental groups (albeit not significantly), which was due to a dose-dependent decrease in the cholesterol content of the low-density lipoprotein (LDL) fraction. These results suggest that dietary xanthoangelol decreases serum LDL levels. 3. In the liver, significant dose-dependent decreases in relative liver liver weight and total triglyceride content were seen in the 0.02 and 0.10 Xan groups. In addition, a significant decrease in total cholesterol content was found in the 0.10 Xan group, which may be due to an elevation of faecal cholesterol excretion in addition to the decrease in liver weight. 4. Investigation of the hepatic mRNA expression of proteins involved in lipid metabolism indicated that there was a significant increase in peroxisome proliferator-activated receptor (PPAR) alpha mRNA expression associated with the tendency for increases in acyl-coenzyme A (CoA) synthetase and acyl-CoA oxidase mRNA expression in the 0.10 Xan group, which may be responsible, at least in part, for the decrease in hepatic triglyceride content in the xanthoangelol-treated rats. In additon, a significant increase in LDL receptor mRNA expression in the 0.10 Xan group may be responsible, at least in part, for the decrease in serum LDL levels in the xanthoangelol-treated rats. 5. In conclusion, dietary xanthoangelol results in a reduction of serum LDL levels and decreases in total cholesterol and triglyceride contents in the liver of SHRSP. These beneficial effects are more effective following consumption of diet containing 0.10% xanthoangelol.

The liver and small intestine play an important role in maintaining cholesterol and bile acid balance within the body. The purpose of this study was to evaluate the effect of bile duct ligation (BDL) on the expression of intestinal and hepatic genes that are important for cholesterol and bile acid metabolism. Rats were allocated to the BDL group or the sham operation group. Blood, liver and small intestine were obtained after 24, 72 and 168 h from both groups. Serum and hepatic lipids were measured by colorimetric assays and hepatic and intestinal mRNA related to lipid metabolism was studied by reverse transcription-polymerase chain reaction (RT-PCR). Hepatic apolipoprotein (Apo) AIV, multidrug resistant protein (Mrp)2, adenosine triphosphate (ATP) binding cassette transporter (Abc)g5 and Abcg8 expression constantly decreased after BDL. Intestinal Apo AIV, Apo CIII, Mrp2, Abcg5 and Abcg8 expression remarkably decreased 24 h after BDL and recovered 72 and 168 h after BDL. Hepatic small heterodimer partner (Shp) expression did not change after BDL. Conversely, intestinal Shp expression remarkably decreased 24 h after BDL (16% of sham operation) and slightly recovered 168 h after BDL (58% of sham operation). Several intestinal mRNA expressions important for lipid (Apo AIV, Apo CIII, Abcg5 and Abcg8) and bile acid (Mrp2 and Shp) metabolism were decreased in the early phase of obstructive jaundice and the expression of these intestinal mRNA recovered in the late phase of obstructive jaundice.

Hemochromatosis represents primary or secondary deposition of an excessive amount of iron in many tissues and organs in the body. Recent studies have evaluated regional myocardial function by tissue Doppler echocardiography. Myocardial strain imaging facilitates the evaluation of regional myocardial function without being influenced by cardiac rotation and translation. Several studies have reported the usefulness of myocardial strain and myocardial strain gradient imaging in patients with various myocardial diseases. In this paper, we performed myocardial strain imaging in a patient with secondary hemochromatosis. © 2007, Japanese Society of Echocardiography. All rights reserved.

Background and Aim: Multidrug resistance protein 2 (Mrp2), Mrp3, adenosine triphosphate-binding cassette transporter g5 (Abcg5) and adenosine triphosphate-binding cassette transporter g8 (Abcg8) have been identified as bile acid or cholesterol transporter in the enterocytes as well as hepatocytes. The purpose of the present study was to evaluate intestinal and hepatic adenosine triphosphate-binding cassette transporter expressions during cholestasis. Methods: Experiment 1: Rats were subjected to bile duct ligation or sham operation. Blood, liver and small intestines were obtained 24 and 72 h after operation. Experiment 2: Rats were divided into four groups as follows: (i) control group; (ii) diosgenin group (fed with diosgenin in diet [1%(wt/wt)] for 7 days); (iii) ethinyl estradiol group (recieved ethinyl estradiol [5 mg/kg daily] for 5 days); and (iv) diosgenin-ethinyl estradiol group (received ethinyl estradiol and diosgenin). After treatment, blood, bile, liver and intestines were obtained. The mRNA related to lipid and bile acid metabolism was analyzed by reverse transcription polymerase chain reaction. Results: Intestinal Mrp2 and Abcg5/Abcg8 mRNA expression remarkably decreased 24 h after bile duct ligation (43% and 61%/54% of sham operation) and recovered 72 h after bile duct ligation (103% and 95%/83% of sham operation). Intestinal Mrp3 mRNA expression did not change after bile duct ligation. Intestinal Mrp2 mRNA expression was remarkably increased in diosgenin and diosgenin-ethinyl estradiol groups in comparison with the control group. There were no significant differences in intestinal Mrp3 mRNA expression among the four groups. Hepatic Mrp3 mRNA expression was remarkably increased in the D, EE and DE groups in comparison with the control group (531%, 321% and 1160% of control, respectively, P < 0.01). Hepatic Abcg5 and Abcg8 mRNA expression decreased in ethinyl estradiol and diosgenin-ethinyl estradiol groups compared with the control group and there were no differences in intestinal Abcg5 and Abcg8 mRNA expressions among the four groups. Conclusion: Bile duct ligation affects not only hepatic but also the intestinal Mrp2 and Abcg5 and Abcg8 expressions. Intestinal Mrp2 mRNA level was regulated by factor in the lumen (e.g. diosgenin feeding). Cholestasis by ethinyl estradiol treatment was enhanced by diosgenin and the increase in hepatic Mrp3 mRNA level might affect the enhancement. (C) 2005 Blackwell Publishing Asia Pty Ltd.

Background and Aim: Multidrug resistance associated gene product 2 (Mdr2) is believed to have a significant role in biliary cholesterol and phospholipid secretions. Both pravastatin and bezafibrate resulted in Mdr2 induction, but increased cholesterol secretion was observed only in pravastatin treatment. To explore the mechanism, the hepatic expression of genes that are responsible for the metabolism of the lipids was studied. Methods: Rats were divided into three experimental groups: (i) the control group; (ii) the bezafibrate group, which was fed a diet containing 0.45% bezafibrate for 5 days; and (iii) the pravastatin group, which was fed a diet containing 0.1% pravastatin for 5 days. Serum, hepatic and biliary lipids were measured by colorimetric assays and hepatic mRNA related to lipid metabolism was studied by reverse transcription-polymerase chain reaction (RT-PCR). Results: In the bezafibrate group biliary phospholipid secretion was increased although cholesterol secretion was not increased. In the pravastatin group, biliary cholesterol and phospholipid secretions were significantly increased. The biliary cholesterol/phospholipid ratio was decreased in the bezafibrate group, but the ratio did not change in the pravastatin group. Hepatic Mdr2, Abcg5 and Abcg8 mRNA expression was remarkably increased in the pravastatin group in comparison with the control group (184%, 264% and 247% of control value, respectively). In the bezafibrate group the hepatic gene expression of Mdr2 was increased (157% of control value), but there were no significant changes in hepatic Abcg5 and Abcg8 mRNA expression compared with the control group. Conclusions: Compared with Mdr2, Abcg5 and Abcg8 seem to be more essential transporters for biliary secretion of cholesterol. Pravastatin upregulated Abcg5/Abcg8 while bezafibrate did not, which appears to explain the different effects of these compounds on biliary lipid secretion. (C) 2004 Blackwell Publishing Asia Pty Ltd.

純化食飼育の Wistar/Crj, WKY/Izm, SHRSP/Izmにおいて、種差間における肝臓脂質代謝の違いを解明するため肝臓脂質代謝関連タンパクmRNA発現量について検討を加えた（英文）。

1. To assess the effect of dietary phytosterol on stroke and the lifespan of salt-loaded stroke-prone spontaneously hypertensive rats (SHRSP), we investigated the effects of the addition of phytosterol to soybean oil (phytosterol content: 0.3%) on stroke onset, lifespan following onset of stroke and overall lifespan compared with canola oil (phytosterol content: 0.9%). 2. Six-week-old male SHRSP were fed a test diet prepared by the addition of canola oil (CA diet), soybean oil (SO diet), soybean oil plus 0.6% phytosterol (SO + 0.06P diet) or soybean oil plus 4.5% phytosterol (SO + 0.45P diet) as a 10% fat source. 3. Systolic blood pressure (SBP) increased in the SO + 0.06P and SO + 0.45P groups compared with the SO group and the increase was dependent on the amount of phytosterol added, indicating that the addition of phytosterol to soybean oil may promote an increase in SBP in salt-loaded SHRSP. 4. The onset of stroke was shortest in the SO + 0.45P group and survival after the onset of stroke was shortest in the CA group. Consequently, the SO + 0.45P and CA groups showed marked lifespan shortening, indicating that a fivefold greater amount of phytosterol was required to produce an effect equivalent to that of canola oil. 5. Investigation of the mRNA expression of ATP-binding cassette (ABC) transporters involved in intestinal phytosterol absorption indicated significant decreases in the intestinal mRNA expression of Abcg5 and Abcg8 in SHRSP and Wistar-Kyoto rats compared with Wistar rats. 6. In conclusion, the addition of phytosterol to soybean oil elevated SBP and promoted the onset of stroke, which may cause a reduction in survival time. However, a fivefold greater amount of phytosterol was required to produce an effect that was equivalent to the survival time-shortening effect of canola oil. The significant decrease in the intestinal mRNA expression of Abcg5 and Abcg8 in SHRSP may be responsible, at least in part, for the unfavourable effects observed following the addition of phytosterol.

In this study, we evaluate the effect of diosgenin, ethinyl estradiol and these co-administration on changes of lipoprotein metabolism and expression of hepatic genes those are important for cholesterol metabolism including the recently identified abcg5 and abcg8 in male Wistar rats. Rats were subjected to four experimental groups: (1) control group, (2) diosgenin group, which was fed the diet containing 1% diosgenin for 7 days, (3) ethinyl estradiol group, which received ethinyl estradiol in a dose of 5 mg/kg daily for 5 days, (4) diosgenin-ethinyl estradiol group, which received ethinyl estradiol treatment and was fed the diet containing 1% diosgenin. Diosgenin-feeding induced the hepatic abcg5/abcg8 expressions and biliary cholesterol secretion. Ethinyl estradiol administration reduced hepatic abcg5/abcg8 expressions and biliary cholesterol secretion. There was a positive correlation between hepatic expressions of abcg5/abcg8 and biliary cholesterol secretion. These findings strongly suggest that abcg5 and abcg8 are key proteins for biliary cholesterol excretion. Diosgenin-feeding did not affect the hepatic abcg5/abcg8 expressions and biliary cholesterol excretion in ethinyl estradiol-treated rat. Serum bile acid and bilirubin were higher and biliary bile acid and bilirubin secretions were lower in diosgenin-ethinyl estradiol group than those in ethinyl estradiol group. This finding suggests that diosgenin enhances the cholestatic effect of ethinyl estradiol in the rat. In conclusion, alteration of biliary cholesterol secretion is related to the expressions of hepatic abcg5 and abcg8 in diosgenin- or ethinyl estradiol-treated rat. (C) 2003 Elsevier Science B.V. All rights reserved.

Platelet-activating factor acetylhydrolase (PAF-AH) is an enzyme that catalyzes hydrolysis of PAF and is important for regulation of serum PAF concentration. Twenty-seven patients with hyperbililubinemic hepatobiliary diseases (eight patients with liver cirrhosis, five with acute hepatitis and 14 with obstructive cholestasis) were studied. Serum PAF-AH activity was elevated in hyperbilirubinemic condition associated with liver cirrhosis, obstructive cholestasis or acute hepatitis. There was a significant correlation of serum PAF-AH activity with serum cholesterol (r=0.891, P=0.014 in liver cirrhosis, r=0.813, P=0.002 in obstructive cholestasis) or low density lipoprotein (LDL) cholesterol (r = 0.771, P = 0.007 in obstructive cholestasis). Serum PAF-AH activity/cholesterol ratio and PAF-AH activity/LDL cholesterol ratio were elevated in the three types of hyperbilirubinemic liver disease. LDL cholesterol increased and PAF-AH/LDL cholesterol ratio decreased in acute hepatitis after remission. On the other hand, LDL cholesterol decreased and PAF-AH/LDL cholesterol ratio did not change after treatment in obstructive cholestasis. These findings suggest that, though serum LDL is a factor influencing serum PAF-AH activity in obstructive jaundice, other factors, i.e. reduced biliary PAF-AH excretion, may also influence serum PAF-AH activity in hyperbilirubinemic hepatobiliary diseases. (C) 2003 Elsevier Science B.V. All rights reserved.

The purpose of this study is to evaluate the effect of bile duct ligation (BDL) on changes of lipoprotein metabolism and hepatic gene expressions that are important for cholesterol metabolism. In male rats serum, very low-density lipoprotein cholesterol, low-density lipoprotein cholesterol and apolipoproteins B and E increased after 5 days of BDL compared with those of sham operation (control) group. Serum apolipoprotein A-IV concentration in the BDL group was lower than that in the control group. In both groups, there was no difference in hepatic lipid concentrations. Hepatic mRNA expressions of scavenger receptor B1, microsomal triglyceride transfer protein (mtp), HMG-coA reductase, cholesterol 7alpha-hydroxylase and multidrug resistance gene product 2 in the BDL group were significantly higher than that in the control group. The adipocyte determination and differentiation I mRNA expression in the BDL group was significantly lower than that in the control group. abcg5 and abcg8 mRNA expressions were remarkably decreased in the BDL group. In conclusion, in obstructive jaundice, metabolism of lipoprotein and proteins that are important for lipid metabolism are drastically changed probably for maintaining the hepatic lipid concentration. The remarkable down-regulation of the abcg5 and abcg8 may be an adaptive change reflecting the inability of biliary cholesterol excretion. (C) 2002 Elsevier Science B.V. All rights reserved.

A 20-item questionnaire was administered to 253 physicians and 111 medical students, who did not have previous clinical clerkship experience, upon completion of their clinical clerkship. Medical students responded that they enjoyed their clinical clerkships but felt pressured and physically tired. Ninety percent of these medical students developed expectations for their career choice during their clinical clerkship. Only 18% of physicians felt that they allowed students enough chance to participate in clinical practice. We must emphasize that the success of any clinical clerkship system depends on an effective communication system between physicians and medical students. © 2003 Elsevier Science Ireland Ltd. All rights reserved.

植物ステロール含量の低い大豆油への植物ステロール添加が、食塩負荷SHRSPの寿命に及ぼす影響、そして、植物ステロール添加効果はSHRSPに特異的であるのかどうか、関連酵素のmRNA発現動態を中心として検討を加えた（英文）。

Regulation of bilirubin glucuronide transporters during hyperbilirubinemia in hepatic and extrahepatic tissues is not completely clear. In the present study, we evaluated the regulation of the bilirubin glucuronide transporters, multidrug resistance-associated proteins (MRP)2 and 3, in rats with obstructive jaundice. Bile duct ligation (BDL) or sham operation was performed in Wistar rats. Liver and kidneys were removed 1, 3, and 5 days after BDL (n = 4, in each group). Serum and urine were collected to measure bilirubin levels just before animal killing. MRP2 And MRP3 mRNA expressions were determined by real-time RT-PCR. Protein expression of MRP2 and MRP3 was determined by Western blotting. Renal MRP2 function was evaluated by para-aminohippurate (PAH) clearance. The effect of conjugated bilirubin, unconjugated bilirubin, human bile, and sulfate-conjugated bile acid on MRP2 gene expression was also evaluated in renal and hepatocyte cell lines. Serum bilirubin and urinary bilirubin excretion increased significantly after BDL. In the liver, the mRNA expression of MRP2 decreased 59, 86, and 82%, and its protein expression decreased 25, 74, and 93% compared with sham-operated animals after 24, 72, and 120 h of BDL, respectively. In contrast, the liver expression of MRP3 mRNA increased 138, 2,137, and 3,295%, and its protein expression increased 560, 634, and 612% compared with sham-operated animals after 24, 72, and 120 h of BDL, respectively. On the other hand, in the kidneys, the mRNA expression of MRP2 increased 162, 73, and 21%, and its protein expression increased 387, 558, and 472% compared with sham-operated animals after 24, 72, and 120 h of BDL, respectively. PAH clearance was significantly increased after BDL. The mRNA expression of MRP2 increased in renal proximal tubular epithelial cells after treatment with conjugated bilirubin, sulfate-conjugated bile acid or human bile. Upregulation of MRP2 in the kidneys and MRP3 in the liver may be a compensatory mechanism to improve bilirubin clearance during obstructive jaundice.

Background. Despite advances in endoscopic treatment methods for upper GI hemorrhage, hemostasis is often difficult to achieve. This study evaluated the usefulness of endoscopic band ligation for upper GI hemorrhage exclusive of hemorrhage from chronic gastroduodenal ulcer and varices. Methods: This prospective study included 27 patients who underwent endoscopic band ligation and 31 patients in whom bipolar electrocoagulation was performed for upper GI hemorrhage. In both groups, the causes of hemorrhage included Dieulafoy's ulcer, Mallory-Weiss tear, gastric ulcer after polypectomy, and gastric angiodysplasia. Patients with esophageal varices and those with chronic gastroduodenal ulcer were excluded. Results: Hemostasis was achieved in all 27 patients in the endoscopic band ligation group and in 26 of 31 patients (83.9%) in the bipolar electrocoagulation group. The median procedure time required for achieving hemostasis was 17.0 minutes (interquartile range: 11.5-23.5) in the endoscopic band ligation group versus 27.0 minutes (interquartile range: 20.5-40.0) in the electrocoagulation group. No major complications occurred in either group. Conclusion: Endoscopic band ligation is efficient, simple, and safe. Therefore, this technique should be considered as a treatment option for nonvariceal, nonchronic gastroduodenal ulcer upper GI hemorrhage.

近畿大学のテュートリアル教育の効果と問題点を学生およびテュータに対してアンケート調査を行い検討した。

Components of the multidrug resistance-associated protein (mrp) family mediate the adenosine triphosphate (ATP)-dependent transport of conjugated organic anions in the liver. Of these, mrp1 and mrp2 hake been shown to have similar substrate specificity and nucleotide sequence, The intracellular localization and distribution of mrp1 under normal condition and cholestasis have not been as yet completely elucidated, To clarify this point, in the present study we evaluated the intracellular localization of mrp1 in rat liver and kidney after bile duct ligation (BDL). Bile duct was ligated in Wistar rats. Sequential staining of mrp1 by immunofluorescence was carried out in rat liver and kidneys 1, 3, and 5 days after bile duct ligation using confocal laser scanning microscopy. Weak granular staining of mrp1 was observed in cytoplasm of control rat hepatocytes. In addition to increased cytoplasm staining of mrp1, belt-and granule-like staining of mrp1 in basolateral membrane of hepatocytes was also shown after BDL. Furthermore, mrp1 immunofluorescence increased over time after BDL. No specific immunoflurescence of mrp1 was detected in control rat kidney. However. mrp1-positive staining was observed in epithelia of some renal tubules after BDL. This study showed that mrp1 immunofluorescence increased in hepatocyte basolateral membrane and cytoplasm and epithelia of some renal tubules after BDL. This increased mrp1 expression may be an adaptive response to impairment of hepato-biliary organic anion transport during obstructive cholestasis. (C) 2002 Elsevier Science Ireland Ltd. All rights reserved.

消化器内視鏡診断・治療の分野においても正確で必要かつ充分な医療情報を提供することは当然のことである.リスクの高い内視鏡検査や治療だけでなく通常の一般内視鏡検査においても文書による説明と拒否を含めた同意が必要とされつつある.従来本院では各科担当医が各々独自に口頭で内視鏡検査・治療の説明を行い, 外科手術用も兼ねた承諾書を使用していたため, その内容にはばらつきがあり必要かつ充分な説明や了解が得られていない例も散見された.時間的制約の多い日常診療において, より効果的に医療情報を提供し患者の充分な理解や自主的な選択を得るためには内視鏡診断・治療専用の説明書と承諾書の使用が適切と考えた.そこで我々は本院独自の説明書と承諾書を新たに作成したので, 現状における問題点や作成上の留意点とその運用などについて報告する.

Kinki University School of Medicine introduced clerkships for undergraduate clinical training in 1999. Clinical clerkships are performed for the first 8 weeks of the sixth academic year. In 1999 and 2000 we conducted questionnaire surveys asking students about this system. The teaching staff encourages students to participate extensively in clinical situations, which reflects the consensus about this system. We also performed similar surveys of nurses and teaching staff. Clinical clerkships did not increase the incidence of problems between patients and medical staff. Many students felt their motivation to be a physician was increased. Although the findings of these questionnaire surveys indicate that our clerkship system works successfully, they also revealed some problems for sixth-year students. Although we recognize the significance of this system for undergraduate clinical training, further improvement is required.

雄性SDラットに総胆管結紮(BDL)を施行し胆汁うっ滞モデルを作製し,肝及び腎におけるmrp1及びmrp2の発現の変化を蛍光免疫染色により検討した.その結果,肝細胞毛細胆管側膜上にmrp2蛋白のdown-regulationが観察された.mrp1は僅かに肝細胞質内に認められたが,BDLにより毛細胆管側膜及び外側膜に発現が認められた

Bilirubin conjugates are secreted from hepatocytes into bile. Monoglucuronosyl bilirubin is an endogenous substrate for the multidrug resistance protein ?, which is located in rat hepatocyte canalicular membrane. We have characterized this ATP-dependent transport using rat canalicular membrane vesicles. Monoglucuronosyl bilirubin, H-3-labeled in the glucuronosyl moiety, was synthesized enzymatically using recombinant UDP-glycosyltransferase 1Al, and was stabilized with ascorbate. The rate for ATP-dependent transport of monoglucuronosyl bilirubin (at 0.5 muM) was 7.3 +/- 1.1 pmol mg protein(-1) min(-1) and the K-m value was 1.3 +/- 0.4 muM. This is the first time to demonstrate this kinetic constant of monoglucuronosyl bilirubin for the rat hepatocyte canalicular membrane. The K-m value is similar to one for recombinant rat multidrug resistance protein 2. (C) 2001 Elsevier Science Ireland Ltd. All rights reserved.

The liver and kidney play a major role in the elimination of a wide variety of endo-xenobiotic anionic substrates into bile and urine. They are mediated by adenosine triphosphate (ATP)-dependent conjugate export pumps, multidrug resistance-associated proteins. In this report, chronological regulations and localization of mrp1 and mrp2 in rat liver and kidney during bile duct ligation were exanimated using immunoflurescence staining. BDL induced down-regulation and fuzzy pericanalicular localization of mrp2 in rat liver. This change was accompanied by the corresponding increase in hepatorenal mrp1, increase in renal mrp2. Reciprocal changes of these transporters in liver and kidney are probably the results of an adaptation to impaired hepatic excretion of conjugated organic anions.

Bilirubin is taken up from blood into hepatocytes by sinosuidal membrane transporters and then excreted into bile through the bile canalicular membrane mainly as bilirubin glucuronides. (1) Mechanism of bilirubin uptake into hepatocytes: many organic anions are incorporated into hepatocytes by organic anion transporting polypeptides (rat, oatp 1, oatp2. oatp3 human, OATP), liver-specific transporter (r1st/HLST), and/or by organic anion transporters (OAT2, OAT3). Oatp1 and HLST transport bilirubin monoglucuronide. However, a transporter of unconjugated bilirubin in the sinusoidal membrane has not as yet been identified. Unconjugated bilirubin may also go across the hepatocyte sinusoidal membrane by a diffusion process. (2) Intrahepatic transport and conjugation of bilirubin: ligandin carries bilirubin to the endoplasmic reticulum (ER) of hepatocytes. In the ER, bilirubin is conjugated by bilirubin uridine diphosphate (UDP)-glycosyltransferase (bilirubin UGT; UGT1A1) to form mono- and diglucuronides of bilirubin. (3) Transport mechanism of bilirubin glucuronides across the hepatocyte canalicular membrane: at the canalicular membrane, bilirubin glucuronides are excreted into bile by multidrug resistance-associated protein 2 (MRP2), a member of the ATP-binding cassette transporter family. (4) Regurgitation of bilirubin glucuronides into blood: MRP3, which is located in the lateral membrane, transports bilirubin glucuronides into blood under conditions of impaired biliary bilirubin excretion.

Background & Aims: Multispecific organic anion-transporting polypeptides (Oatps) are involved in the transcellular movement of amphipathic compounds in many tissues including the liver, kidney, and blood-brain barrier. Recently, a high-affinity digoxin transporter (Oatp2) was cloned from rat brain and shown to be also expressed in the liver. Methods: We investigated the cellular and subcellular distribution of Oatp2 in rat liver by in situ hybridization technology and immunofluorescence microscopy and compared its substrate specificity with that of Oatp1 in complementary RNA-injected Xenopus laevis oocytes. Results, The results show a selective basolateral (sinusoidal) expression of Oatp2 in midzonal to perivenous hepatocytes, but not in periportal or the innermost layer of perivenous hepatocytes. Common substrates of both Oatp1 and Oatp2 include bile salts, steroid conjugates, thyroid hormones (T3, T4), ouabain, and the endothelin receptor antagonist BQ-123 (Michaelis constants: Oatp1, similar to 600 mu mol/L; Oatp2, similar to 30 mu mol/L). Other organic anions including sulfolithotaurocholate, bilirubin monoglucuronide, and sulfobromophthalein were transported only by Oatp1. Conclusions: These results provide definite evidence for the partially overlapping and partially selective substrate specificities of Oatp1 and Oatp2. The unique acinar distribution of Oatp2 might indicate that it represents a high-affinity "backup'" system for complete hepatocellular removal of certain cholephilic substances from portal blood plasma.

The secretion of bilirubin conjugates from hepatocytes into bile represents a decisive step in the prevention of hyperbilirubinemia. The bilirubin conjugates, monoglucuronosyl bilirubin (MGB) and bisglucuronosyl bilirubin (BGB), were previously suggested to be endogenous substrates for the apical multidrug resistance protein (MRP2), a member of the adenosine triphosphate (ATP)-binding cassette family of transporters (symbol ABCC2), also termed canalicular multispecific organic anion transporter. We have characterized this ATP-dependent transport using membrane vesicles from human embryonic kidney (HEK) cells expressing recombinant rat as well as human MRP2, MGB and BGB, H-3-labeled in the glucuronosyl moiety, were synthesized enzymatically with recombinant UDP-glucuronosyltransferase 1A1, and stabilized with ascorbate, Rates for ATP-dependent transport of MGB and BGB (0.5 mu mol/L each) by human MRP2 were 183 and 104 pmol x mg protein(-1) x min(-1), respectively. K-m values were 0.7 and 0.9 mu mol/L for human MRP2, and 0.8 and 0.5 mu mol/L for rat MRP2, with MGB and BGB as substrates, respectively. Leukotriene C-4 and 17 beta-glucuronosyl estradiol, which are both known high-affinity substrates for human MRP2, inhibited [H-3]MGB transport with IC50 values of 2.3 and 30 mu mol/L, respectively. Cyclosporin A competitively inhibited human and rat MRP2-mediated transport of [H-3]MGB, with K-i values of 21 and 10 mu mol/L, respectively, Our results provide direct evidence that recombinant MRP2, cloned from rat as well as human liver, mediates the primary-active ATP-dependent transport of the bilirubin conjugates MGB and BGB.

Many organic compounds are taken up from the blood by membrane transporters, taken across the sinosuidal membrane of hepatocytes and then excreted into bile via the bile canalicular membrane. The hepatic uptake of conjugated bile acids is mediated by the sodium taurocholate cotransporting polypeptide. Many organic anions and bulky organic cations are incorporated into hepatocytes by the organic anion transporting polypeptide, while small organic cations are transported by the organic cation transporter. Ar the canalicular membrane, organic compounds are excreted into bile by ATP-binding cassette transporters which hydrolyse ATP to ADP. Excretion of monovalent bile acids is mediated by the canalicular bile salt transporter and that of organic anions, including divalent bile acid, conjugates, are mediated by the multi-drug resistance-associated protein 2, also termed canalicular multi-specific organic anion transporter. Organic cations are excreted into bile by the multi-drug resistance gene product (MDR) 1 and phospholipids are excreted by MDR3 (mdr2 in mice and rats). The clinical syndromes associated with alterations of these transporters are also discussed. (C) 1999 Blackwell Science Asia Pty Ltd.

Backgrounds/Aims: No study has so far been conducted to clarify whether the presence of hyperbilirubinemia is detrimental to liver and renal functions. In the present study, the effects of polyethylene glycol-modified bilirubin oxidase (PEG-BOX) therapy on liver and renal function tests, hepatic energy charge and urinary prostaglandin levels mere evaluated in a rat model of obstructive jaundice. Methods: Sprague-Dawley rats were used in the experimental model of obstructive jaundice. PEG-BOX or an equivalent amount of PEG alone was intravenously injected into the animals and sampling of blood and urine, and liver harvesting mere done sequentially after bile duct ligation. Results: Conventional liver function tests showed no difference between PEG-BOX and control groups. However, bilirubin concentrations in the peripheral blood and liver tissue specimens markedly decreased, and the hepatic energy charge significantly increased in the PEG-BOX group as compared to controls. The blood concentration of bile acid was lower, but its urinary excretion was higher in the PEG-BOX group than in the control group. In vitro incubation of PEG-BOX with serum from rats with obstructive jaundice decreased the concentration of bilirubin but not that of bile acid. The urinary levels of prostaglandin E2 and the thromboxane B2/6-keto-prostaglandin F1 alpha ratio were significantly lower in the BEG-BOX group than in the control group. Conclusions: The systemic reduction of bilirubin concentration may contribute to normalization of the urinary levels of prostaglandins and thromboxane B2, to decrease in serum bile acid levels, and to improvement of the hepatic energy charge in obstructive jaundice, These findings suggest that preoperative improvement of jaundice may be beneficial to patients with obstructive jaundice.

Crigler-Najjar syndrome (CN) type II is caused by a reduction in hepatic bilirubin uridine 5′-diphosphate (UDP)-glucuronosyltransferase activity. Recently, there has been progress in mutation analysis of patients with CN type II. Here, we analyzed both the coding and the promoter regions of the gene in seven Japanese patients with CN type II from five unrelated families. The mutations found in this study were classified into three types. The first type was composed of double homozygous missense mutations (Gly71Arg and Tyr486Asp) in exons 1 and 5. These mutations, which were detected in five patients from three unrelated families, were the commonest. The second type, which was detected in one patient, consisted of a single homozygous missense mutation (Arg209Trp) in exon 1. The third type, which was detected in one patient and was a new type of mutation combination, was composed of a homozygous insertion mutation of the TATAA element and a heterozygous missense mutation (Pro229Gln) in exon 1. Although the first and the second type of mutations are recessive, the third type appears to be dominant with incomplete penetrance, since the allele frequency of the insertion mutation of the TATAA element is very high (40%).

体質性黄疸は肝細胞の先天的なビリルビン代謝異常により発症する黄疸で,高間接ビリルビン血症のGilbert症候群, Crigler-Najjar症候群I, II型と高直接ビリルビン血症のDubin-Johnson症候群, Rotor症候群に分類される.近年,高間接ビリルビン血症の3症候群が同一遺伝子の異常による事が明らかとなり,又Dubin-Johnson症候群についても毛細胆管側肝細胞膜上の有機アニオン輸送蛋白の欠損が判明し,病態の理解が飛躍的に進展した.

Transport of various intrinsic and extrinsic organic anions across the canalicular membrane of hepatocytes has been confirmed to occur via ATP-dependent primary active transport. To investigate these processes in the presence of obstructive jaundice, the uptake of radiolabeled taurocholate and pravastatin, a non-bile acid organic anion, by isolated hepatocyte canalicular membrane vesicles was observed after bile duct ligation in rats. The Mg++-ATPase and Na+, K+ -ATPase activity did not differ between membrane vesicle preparations from sham-operated and bile duct-ligated rats. Taurocholate uptake decreased significantly to 45% and 29%, while pravastatin uptake decreased to 35% and 35% of that in sham-operated rats at 2 and 7 days after bile duct ligation. These results indicate that the ATP-dependent transport systems for both bile salts and non-bile acid organic anions are impaired soon after the onset of obstructive jaundice.

To elucidate the possible implication of hepatic blood supply to the occurrence of hepatolithiasis, the rat superior mesenteric vein, which drains blood from the intestine, or the splenic vein, which drains from the spleen, was occluded for 30 minutes. Changes in the hepatic oxygen saturation index, intrahepatic uridine diphosphate-glucuronic acid concentration, bile flow, and the excretion of bilirubin as well as its fraction, along with bile acid in bile before and after the procedure, were observed. in association with superior mesenteric vein occlusion, oxygen saturation index, hepatic uridine diphosphate-glucuronic acid concentration, bile flow, bile acid concentration in bile, and percentage of biliary bilirubin diglucuronide were all decreased. Incubation of bile under sterile conditions from rats with occluded superior mesenteric veins resulted in precipitation of mainly calcium salt of fatty acid. In contrast, splenic vein occlusion caused no changes except for a decrease in biliary bilirubin concentration. Incubation of bile from rats with occluded splenic veins did not induce precipitation. From these findings it can be concluded that blood flow in the superior mesenteric vein is the primary source of oxygen supply to the rat liver and that this vein plays an important role in maintaining bile flow, bile acid excretion, and bilirubin conjugation and in preventing the precipitation of bile (possibly preventing hepatolithiasis).

肝疾患の診断に関する著書。

胆汁うっ滞に関する最近1年間の基礎的，臨床的研究の進展を概説した．

体質性黄疸の成因、病態生理、臨床像について解説した。

肝臓の重要な機能である胆汁分泌のメカニズムについて概説した。

血中ビリルビンの測定法及びその測定の臨床的意義を臨床医向けに概説した。

溶血性黄疸に対する治療方針を概説した。

電子カルテの導入を視野に入れ、日常よく見られる疾患であるＣ型慢性肝炎の臨床評価モデルを作成した。

一般内科医にとって診療録のあり方を、我が国の医療環境の変化を考慮に入れ論じた。

各種体質性黄疸に関し、その生検像、電顕像、画像を提示し、各々について解説した。

ビリルビン代謝異常を呈する疾患を解説した。

ビリルビンの測定法とその臨床意義について概説した。

ヘモクロマトーシス発症の分子メカニズムを解説した。

Wilson病発症の分子メカニズムを解説した。

近畿大学薬学部・医学部合同学習会におけるPBL教育に関する報告。

ラットに閉塞性胆汁うっ滞を実験的に作成し肝臓と小腸における脂質および胆汁酸代謝関連遺伝子産物の発現の経時的変化を観察した．閉塞性胆汁うっ滞時には従来から知られている肝臓におけるこれらの変化のみならず小腸においての発現もダイナミックに変化していた．

SHR系ラットに特有な脂質代謝を解明することを目的として、ウィスターラットを対照とし、種差間における脂質代謝の違いを小腸での脂質代謝関連遺伝子発現動態を中心として検討を加え、種特異的に小腸におけるmRNA発現量の違いが有ることを明らかにした。

II型糖尿病のモデル動物として知られ、糖尿病発症を段階的に捉えることが可能なOLETF ラットに着目し、インスリン抵抗性未発症の若齢期におけるOLETFラットの脂質代謝ならびに高コレステロール食投与による反応性高コレステロール血症について、LETOラットと比較検討した。OLETFラットでは、インスリン抵抗性未発症の若齢期より脂質代謝異常が存在し、高コレステロール食投与により顕著な反応性高コレステロール血症を示すことが明らかとなった。

Bezafibrateとpravastatinはともにラットにおいて胆汁中コレステロール排泄を促進させる。この現象を肝細胞におけるコレステロール輸送蛋白質であるAbcg5／Abcg8の発現から論じた。

若齢期OLETF ラットにコレステロール食を投与すると、LETOラットと比較してVLDL, LDL両分画の顕著な上昇とapoA-I, apoE含量の減少を伴ったHLD分画の大きな減少が認められた。

OLETFラットでは、LETOラットと比較して血清コレステロール、リン脂質、遊離脂肪酸、apoA-I, apoB, apoE含量が有意に高値を示し、これらはLDL及びHDL分画の上昇を示唆していた。OLETFラットでは、インスリン抵抗性未発症の若齢期より血清リポ蛋白代謝異常が存在することが明らかとなった。

２型糖尿病のモデルラットであるOLETFラットと対照動物であるLETOラットについて、まだ糖尿病を発症していない若齢期に高脂質食を摂取させた際の脂質代謝の違いを比較検討した。

２型糖尿病のモデルラットであるOLETFラットと対照動物であるLETOラットについて、まだ糖尿病を発症していない若齢期における脂質代謝の違いを比較検討した。

Wistar/Crj, WKY/Izm, SHRSP/Izm の比較検討から、SHR系統特有な肝臓ステロール代謝関連蛋白のmRNA発現量の違いがあることを報告した。

ペリラ油への植物ステロール添加は食塩負荷SHRSPの血圧上昇を促進し、卒中発症・寿命を短縮した。小腸Abc (ATP-binding cassette transporter) g5, g8 及び肝臓Add1（Adipocyte determination and differentiation protein 1）の関与が示唆された。

植物コレステロールの一種であるジオスゲニンは、胆汁中にコレステロール排泄増加をきたすが、この機序として新しく発見されたコレステロール輸送蛋白の発現の増加が関与している事を示した。

食塩負荷SHRSPの卒中発症・寿命に対するペリラ油の延長作用に対し、植物ステロールは添加量依存的にこれらを短縮した。

純化食飼育の Wistar/Crj, WKY/Izm, SHRSP/Izm において、種差間における血清・肝臓脂質含量の違いが認められた。そしてこれは肝臓における脂質代謝関連タンパクmRNA発現量の違いと密接に関連することが示唆された。

植物ステロール添加は、濃度依存的に食塩負荷SHRSPの寿命を短縮することが明らかとなった。また、SHRSPではABC (ATP-binding cassette transporter) G5、G8等の遺伝子発現動態が異なっていた。

EHBRにおいてはSDラットに比して、肝細胞、腎細胞ともmrp3の発現が増強していた。

HepG2およびRPTECに、bilirubin、bilirubinditaurate,taurolithocholateを添加し、MRP2mRNAの変化を観察した。これらの添加にてMRP2発現は増強が認められた。

閉塞性胆汁うっ滞時には、各種脂質関連遺伝子発現は、1)VLDLアセンブリーの増加、2)胆汁中へのコレステロール排泄の低下に関与する変化が認められた。特に、abcg5とabcg8にては、遺伝子発現は、各々正常の３％、７％まで著減していた。

植物ステロール含量の低い大豆油への植物ステロール添加が、食塩負荷SHRSPの寿命に及ぼす影響、そして、植物ステロール添加効果はSHRSPに特異的であるのかどうか、関連酵素のmRNA発現動態を中心として検討を加えた。

胆管閉塞時のビリルビン排泄機能の変化を、肝臓及び腎臓でのビリルビン輸送蛋白であるMrp2とMrp3の遺伝子レベル、蛋白レベルにおける変動にて検討した。

胆管閉塞ラットにおけるmrp1の組織分布と細胞内局在を検討した。

閉塞性黄疸モデルラットにおけるmrp２の肝臓、腎臓における細胞内局在と腎からのビリルビン排泄機序を検討した。

近畿大学医学部教育における、クリニカルクラークシップの導入後の効果と問題点を、学生、教員、看護職員に対するアンケート調査結果から検討した。

ラット胆管閉塞によって肝細胞膜細胆管側に局在するmrp2は、発現量が低下し、その局在はcytosolに移動した。またmrp1は、類洞側に局在し、発現は増加した。腎においては、尿細管上皮細胞管腔側のmrp2はその発現が増大していた。

肝細胞および尿細管上皮膜上に存在mrp2の閉塞性黄疸時の変化を検討した。閉塞性黄疸時には、肝におけるmrp2の発現は、蛋白レベル、RNAレベルとも低下し、腎においては、両者とも増加していた。

ヒトにおける黄疸発症機構の解明の為、ヒトMRP2およびMRP3の肝細胞における局在、ビリルビン抱合物の輸送実験を行い、その結果、ヒトMRP3が黄疸発症に重要な役割を果たしている事が、あきらかとなった。

肝細胞癌を対象に超音波造影剤Levovist静注によるFundamental Color Doppler法とともにIntermittent Color Doppler法による血流シグナルの増強効果の評価を行い画像上腫瘍の染影効果が認められたので報告した。

植物ステロールにはコレステロール(CHOL)低下作用があることが知られている。一方で過剰になると動脈硬化や胆汁鬱滞性肝障害など生体への毒性が認められる。しかしながら、これらの効能や毒性に対する小腸の役割は十分に解明されていない。近年、血中コレステロールの排泄経路として肝臓から胆管を介した経路だけでなく、新規の排泄経路として腸管に直接排泄する経路であるTICE(trans-intestinal cholesterol efflux)が発見された。そこで、2021年度は食餌中の植物ステロールの含量の異なる油脂を摂取させたラットにおいてSham手術と総胆管結紮(BDL)術を施行し、肝臓から胆汁中へ植物ステロールを排泄するルートを遮断することでTICEに如何なる影響を及ぼすか観察するため以下の研究を行った。方法と結果：6週令雄性ラットにAIN-93に準拠して作成した以下の4群の食餌を摂取させた。1)10%大豆油(S群) 2)10%大豆油+0.005%エゼチミブ(S+E群) 3)10%魚油(F群) 4)10%魚油+0.005%エゼチミブ(F+E群)。各々の飼料で4週(10週令)飼育した後にSham術とBDLを施行し72時間後に実験に供した。BDLにより①血清CHOLの上昇を認め、②TICEを反映する糞中CHOLはS群とF+E群で増加したがS+E群とF群では増加しなかった。③肝Abca1はS群で増加したがS+E群とF群では増加しなかった。④小腸Abcg5とAbcg8はS群で増加したがエゼチミブ添加によりAbcg8の誘導は抑制された。一方、魚油群では誘導は認められなかった。また、TICEに関連している可能性があるLDLレセプターやSR-B1では差は認められなかった。これらの結果からBDL下では大豆油に含まれる毒性物質の蓄積から生体を防御するためにTICEが増強している可能性が示唆された。

Any excessive cholesterol is transported back to the liver from the peripheral tissues and excreted in bile. In addition to the biliary route, transintestinal cholesterol efflux (TICE), a mechanism wherein cholesterol enters the small intestinal epithelial cells from the blood and is directly excreted into the intestinal lumen, was recently reported as a route of cholesterol excretion. In the present study, drug-induced cholestasis and bile duct ligation (BDL) induced intestinal expression of LDL receptor as well as Abcg5/g8 and enhanced fecal cholesterol excretion. Similarly, TICE pathway can be stimulated by lipid-lowering drugs ezetimibe and pemafibrate by increasing intestinal cholesterol efflux transporters Abcg5/g8. Furthermore, pemafibrate may suppress intestinal cholesterol absorption by reducing NPC1L1 and SR-B1. These data indicated that the small intestine may be a target organ for the prevention of atherosclerosis.

Small intestine has a pathway for the excretion of cholesterol from blood into the lumen (TICE). In the present study, we investigated whether the TICE is involved in the cholesterol-lowering effects of fish oil and ezetimibe, in rodent. Fish oil increased faecal cholesterol excretion by 41%, and increased the expression of Abcg5/g8 in the small intestine, which is thought to be associated with faecal cholesterol excretion, suggesting that fish oil may be associated with TICE. Ezetimibe did not increase intestinal Abcg5/g8 expression or faecal cholesterol excretion, but increase hepatic Abcg5/g8 expression. The combination of fish oil and ezetimibe markedly improved fatty liver. The improvement was associated with increased hepatic Abcg5/g8 and Mdr2 expression. These results suggest that fish oil promotes TICE and ezetimibe promotes cholesterol excretion from the liver into bile. The combination was efficient for the reduction of hepatic lipid.

Oxidative stress has been implicated in the transition from simple hepatic steatosis to nonalcoholic steatohepatitis (NASH). The Nuclear factor E2-related factor 2 (Nrf2) transcription factor serves as a cellular sensor for oxidative and electrophilic stress. Nrf2 deletion dysregulates hepatic mRNA expression of lipid and iron metabolism related genes and tends to increase hepatic triglycerides and iron, suggesting that Nrf2 may play a role in the pathogenesis of NASH. Wasabi derivative 6-methylsulfinylhexyl isothiocyanate, a potent Nrf2 activator, prevented the fatty liver produced by a high-fat diet in mice. These data indicated that Nrf2 could prevent the onset and progression of steatohepatitis.