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论文摘要

缺氧诱导大鼠肝星状细胞激活及表达内脂素的实验研究

Effect of Hypoxia on the Activation and Visfatin Expression in Rat Hepatic Stellate Cells

作者:易智慧, 门若庭, 袁聪等

Author:YI Zhi-hui, MEN Ruo-ting, YUAN Cong. et al

收稿日期:          年卷(期)页码:2014,45(4):563-566

期刊名称:四川大学学报(医学版)

Journal Name:JOURNAL OF SICHUAN UNIVERSITY (MEDICAL SCIENCE EDITION)

关键字:肝星状细胞 缺氧诱导因子-1α 内脂素 α-平滑肌肌动蛋白

Key words:Hepatic stellate cells Hypoxia-inducible factor-1α Visfatin α-smooth muscle actin

基金项目:

中文摘要

目的 通过观察常氧及缺氧状态下大鼠肝星状细胞(HSCs)表达α-平滑肌肌动蛋白(α-SMA)、缺氧诱导因子-1α(HIF-1α)和内脂素水平的变化,探讨缺氧对HSCs激活及表达内脂素的影响。方法 对原代大鼠HSCs进行缺氧(37 ℃、5% CO2+1% O2 + 94% N2)处理,检测不同缺氧时间(3、6、12及24 h)和常氧条件(5%CO2+21%O2+74%N2)下HSCs中HIF-1α、α-SMA和内脂素的mRNA表达水平及蛋白表达水平。基因表达用RT-PCR检测,蛋白表达用蛋白质印迹法(Western blot) 检测。结果 缺氧3 h即能诱导大鼠HSCs表达HIF-1α mRNA(Pα-SMA mRNA及蛋白表达水平的表达上调(PPPr=0.991,蛋白r=0.968,P

英文摘要

Objective To investigate the effect of hypoxia on the visfatin and the expression of smooth muscle-actin (α-SMA) and hypoxia-inducible factor-1α (HIF-1α) in rat hepatic stellate cells (HSCs). Methods Rat primary HSCs were isolated from SD rats by in situ perfusion of collagenase and pronase and single-step Nycodenz density gradient centrifugation, and then cultured and activated. Completely activated primary HSCs were exposed to hypoxic conditions (37 ℃, 5% CO2, 1% O2,94%N2), or normoxic conditions (37 ℃, 5% CO2, 21% O2,74% N2), for 3, 6, 12 or 24 h respectively. The expression of α-SMA, the marker of HSC activation, and visfatin were assessed by Real time-PCR and Western blot. The Expression of HIF-1α was detected by Real time-PCR. Results HIF-1α mRNA in rat HSCs was induced after exposed to hypoxia for 3 h, and maintained elevated status up to 24 h. HSCs exposed to 1% O2 hypoxic conditions for 6 h increased α-SMA mRNA and protein expression. Visfatin mRNA expression was up-regulated after subjected to hypoxia for 12 h, and protein level was elevated after 6 h hypoxia. A positive linear correlation existed between α-SMA and visfatin expression in responsible to hypoxia 〔r=0.991 (genes) and r=0.968 (proteins),P

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