葡萄糖和胰岛素调节三明治肝细胞中胆酸相关转运体表达及功能

中国临床药理学与治疗学 ›› 2013, Vol. 18 ›› Issue (8): 847-852.

葡萄糖和胰岛素调节三明治肝细胞中胆酸相关转运体表达及功能

赫蕾¹, 郭岑¹, 姚丹², 盛晶晶², 潘国宇², 刘晓东¹

¹中国药科大学药物代谢动力学重点实验室,南京 210009,江苏;
²中科院上海药物研究所药物安全评价中心,上海 201203

收稿日期:2013-03-02 修回日期:2013-07-09 出版日期:2013-08-26 发布日期:2013-07-31
通讯作者: 刘晓东,男,博士,教授,博导,研究方向:药物转运体和代谢酶。Tel: 025-83271006 E-mail: xdliu@cpu.edu.cn; 潘国宇,男,博士,教授,研究方向:药物代谢和毒性。Tel: 021-20231000 E-mail: gypan@cdser.simm.ac.cn
作者简介:赫蕾,女,硕士在读,研究方向:药物转运体。Tel: 13916488515 E-mail: heheleilei126@126.com
基金资助:
国家自然科学基金面上项目“新型体外模型研究糖尿病中肝脏胆酸药物转运体功能变化”(81173116)

Effects of glucose and insulin on expression and function of bile acid transporters in sandwich cultured rat hepatocytes

HE Lei¹, GUO Cen¹, YAO Dan², SHENG Jing-jing², PAN Guo-yu², LIU Xiao-dong¹

¹China Pharmaceutical University, Drug Metabolism and Pharmacokinetics, Nanjing 210009,Jiangsu, China;
²Shanghai Institute of Materia Medica, Center for Drug Safety Evaluation and Research, Shanghai 201203, China

Received:2013-03-02 Revised:2013-07-09 Online:2013-08-26 Published:2013-07-31

摘要/Abstract

摘要： 目的: 测定葡萄糖、胰岛素等糖尿病血糖中的异常因素对体外培养肝细胞中胆汁酸相关转运体表达及功能的影响。方法: 分离并用三明治法培养大鼠原代肝细胞,用不同浓度葡萄糖或胰岛素培养 72 h,使用胆盐类似物Cholyl-lysyl-fluorescein(CLF)进行摄取转运体钠/胆汁酸共转运体(Na⁺/bile acid cotransporter, ntcp)及外排转运体胆盐外排泵(Bile salt export pump, bsep)转运体功能学实验,并用荧光定量PCR法测定胆汁酸相关转运体ntcp、bsep及调节胆汁酸代谢转运的核受体(Farnesoid X receptor,fxr)的mRNA表达变化。结果: 高浓度的葡萄糖和胰岛素均使CLF的胆泡排泄减少。高糖和高胰岛素均能够降低ntcp的表达(3~4倍),增加bsep的表达(约2倍),高糖能够降低fxr的水平(约 0.5 倍),高胰岛素增加fxr的水平(约 1.5 倍)。结论: 葡萄糖和胰岛素水平异常均会对胆汁酸相关转运体表达和功能造成影响;三明治原代肝细胞可用于胆酸和药物肝胆分布研究。

关键词: 葡萄糖, 胰岛素, 三明治肝细胞, 胆汁酸转运体

Abstract: AIM: To explain the enhancement of bile acid excretion in diabetes from bile acid transporter perspective.METHODS: Primary rat hepatocytes were isolated and cultured in a sandwich configuration (SCRH). Then hepatocytes were cultured with different concentrations of glucose and insulin, and function of Na⁺/bile acid cotransporter (ntcp) and bile salt export pump (bsep) were tested with Cholyl-lysyl-fluorescein (CLF) as a typical substrate. mRNA levels of ntcp, bsep and lipid regulating nuclear receptor fxr were determined with Real Time PCR.RESULTS: CLF biliary excretion was both decreased in high concentrations of glucose and insulin significantly.Both high concentrations of glucose and insulin can decrease ntcp mRNA for 3-4 fold, but increase bsep mRNA for approximately 2 fold. Levels of fxr mRNA was down regulated in high concentrations of glucose and up regulated in high concentrations of insulin.CONCLUSION: Abnormal glucose and insulin concentration have impacts on bile acid transporters in sandwich cultured rat hepatocytes.Sandwich cultured rat hepatocytes could offer a useful tool to explore hepatobiliary disposition of bile acid and drugs.

Key words: Glucose, Insulin, Sandwich cultured rat hepatocyte, Bile acid transporter

中图分类号:

R965.2

赫蕾, 郭岑, 姚丹, 盛晶晶, 潘国宇, 刘晓东. 葡萄糖和胰岛素调节三明治肝细胞中胆酸相关转运体表达及功能[J]. 中国临床药理学与治疗学, 2013, 18(8): 847-852.

HE Lei, GUO Cen, YAO Dan, SHENG Jing-jing, PAN Guo-yu, LIU Xiao-dong. Effects of glucose and insulin on expression and function of bile acid transporters in sandwich cultured rat hepatocytes[J]. Chinese Journal of Clinical Pharmacology and Therapeutics, 2013, 18(8): 847-852.

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