鹅去氧胆酸对小鼠肝灌流及原代肝细胞胆汁酸代谢通路相关基因表达的影响  

Effects of chenodeoxycholic acid on gene expression of bile acid metabolic pathway in liver perfusion and primary hepatocyte

作　　者：鲁艳柳 杨艳萍 杜艺玫 何芋岐 陆远富 Lu Yanliu;Yang Yanping;Du Yimei;He Yuqi;Lu Yuanfu(Key Laboratocy of Basic Pharmacology of Ministry of Education and Joint International Research Laboratory of Ethnomedicine of Ministry of Education,Zunyi Medical University,Zunyi Guizhou 563099,China;The Pharmaceutical Lab,School of Pharmacy,Zunyi Medical University,Zunyi Guizhou 563099,China)

机构地区：[1]遵义医学院基础药理教育部重点实验室暨特色民族药教育部国际合作联合实验室,贵州遵义563099 [2]遵义医学院药学院药学实验室,贵州遵义563099

出　　处：《遵义医学院学报》2018年第4期403-407,共5页Journal of Zunyi Medical University

基　　金：国家自然科学基金资助项目(NO:81660685;81560673;81760678);教育部春晖计划项目(NO:Z2016003);贵州省科技重大专项(NO:黔科合重大专项字[2015]6010);贵州省科学技术基金资助项目(NO:黔科合JZ字[2015]2010;黔科合J字[2015]2158);贵州省教育厅自然科学研究项目(NO:黔教合KY字[2015]373);贵州省留学人员科技创新项目(NO:黔人[2015]03);遵义医学院优秀青年人才项目(NO:15zy-004)

摘　　要：目的通过小鼠原代肝细胞、离体肝灌流模型和整体动物实验,比较鹅去氧胆酸(chenodeoxycholic acid,CDCA)对胆汁酸代谢通路的影响。方法实验包括原代肝细胞、离体肝灌流和整体动物实验3部分,每部分均包括空白组和CDCA给药组(n=6),给药剂量分别为100μmol/L、100μmol/L和50 mg/kg。采用实时荧光定量PCR检测肝细胞或肝脏中Cyp7a1、Cyp8b1、Fxr、Shp、Bsep、Ostβ等基因表达的水平。结果在原代肝细胞的实验中,CDCA能显著上调Shp、Ostβ基因的表达(P<0.05),抑制Cyp7a1、Cyp8b1和Bsep基因的表达(P<0.05)。在离体肝灌流的实验中,CDCA能诱导Ostβ的基因表达(P<0.05),而在整体动物实验中,CDCA对Bsep基因的表达产生明显抑制(P<0.05)。结论由实验结果可知,在肝灌流及原代肝细胞中,CDCA对胆汁酸代谢通路的影响不同,在离体肝灌流模型与整体动物上的影响较一致。因此,在其开发与研究中,不能局限于体外细胞模型,还需要进一步的验证。Objective To compare the effects of chenodeoxycholic acid（CDCA） on bile acid metabolic pathway in the primary hepatocytes,the liver perfusion model and the intact animal. Methods The investigation was completed based on the models of the primary hepatocytes,the perfused liver and the intact animal. Each consisted of the blank group and CDCA group（n = 6）. And the dosages were 100 μmol/l,100 μmol/l and 50 mg/kg respectively. The mRNA of Cyp7a1,Cyp8b1,Fxr,Shp,Bsep,and Ostβ in liver or cells were quantified by real time PCR. Results In the primary hepatocytes,Cyp7a1,Cyp8b1 and Bsep mRNA expressions were reduced obviously（P〈0. 05） by CDCA treatment. And the gene expressions of Shp and Ostβ were up-regulated（P〈0. 05）.Compared with the blank group,CDCA induced the gene expression of Ostβ in the perfused liver（P〈0. 05） and decreased Bsep mRNA in the intact animal（P〈0. 05）. Conclusion The in vitro regulation of the bile acid metabolic pathway by CDCA is different from the in vivo results. The liver perfusion model would be more consistent with the in vivo situation. The efficacy of CDCA in activating its receptors used in vitro cell cultures should be necessarily reproduced in vivo.

关 键 词：鹅去氧胆酸 胆汁酸代谢通路 原代肝细胞 离体肝灌流模型 小鼠 

分 类 号：R965.1[医药卫生—药理学]