乙醇诱导肝星状细胞凋亡模型的建立  

Establishment of ethanol-induced apoptosis in hepatic stellate cell

作　　者：张春燕[1] 陈川宁[1] 陶忠桦 甘淋[1] 

机构地区：[1]西南医科大学基础医学院生物化学与分子生物学教研室,四川泸州646000 [2]西南医科大学公共卫生实验教学中心,四川泸州646000

出　　处：《西南医科大学学报》2017年第2期101-104,共4页Journal of Southwest Medical University

基　　金：国家自然基金项目(No:81341121);四川省卫生厅项目(No.120366);泸州市科技计划项目(泸市科函[2011]42号文件)

摘　　要：目的:探讨乙醇诱导肝星状细胞(hepatic stellate cell,HSC)凋亡模型的作用及其与凋亡相关基因caspase-3、p53、bax和bcl-2表达的关系。方法:用CCK-8法检测不同浓度(0、10、20、40、60和80 m L/L)的乙醇对HSC的杀伤作用。采用40 mL/L乙醇作用6 h诱导HSC细胞凋亡。通过流式细胞术分析乙醇诱导对凋亡的影响,并用实时定量PCR检测乙醇诱导对凋亡相关基因caspase-3、p53、bax和bcl-2表达的影响。结果:随着乙醇浓度的增加,对HSC的杀伤能力逐渐增加。40 m L/L乙醇使HSC发生显著的细胞凋亡变化。流式细胞术检测发现乙醇诱导可使HSC早期+晚期凋亡明显增加(8.76%±0.43%vs 11.51%±0.54%),差异具有统计学意义(t=2.11,P<0.05);促凋亡基因caspas-3、bax和p53的mRNA表达水平上调,抑制凋亡基因bcl-2的mRNA表达水平下调。结论 :低浓度乙醇可诱导HSC凋亡增加,其凋亡发生与凋亡相关基因caspase-3、p53、bax和bcl-2表达改变相关。Objective： To establish ethanol-induced apoptosis in hepatic stellate cell（HSC） and analyze the expression of apoptosis-related genes, caspase-3, p53, bax and bcl-2. Methods： HSC was exposed to different concentrations of ethanol（0、10、20、40、60 and 80 mL / L） for 6 h, and the cytotoxic effect was analyzed by CCK8 assay. Then HSC was treated with 40 mL/L of ethanol for 6 h, apoptosis was analyzed by FACS, and expression of apoptosis-related genes （caspase-3, p53, bax and bcl-2） was evaluated by real-time PCR. Results： The cytotoxic effect of ethanol is directly correlated with its concentration. At 40 mL/L, ethanol induced significant apoptosis in HSC as evaluated by FACS, and significantly increased the expression of caspase-3, p53, and bax, and significantly decreased the expression of bcl-2 as evaluated by real-time PCR. Conclusion： Low dose of ethanol can induce apoptosis in HSC, which may be associated with the altered expression of caspase-3, p53, bax and bcl-2.

关 键 词：乙醇 酒精性肝病 肝星状细胞 凋亡 凋亡相关基因 

分 类 号：R575.2[医药卫生—消化系统]