蛋白酶体抑制剂MG132诱导人血管内皮细胞凋亡及其机制  

Mechanism Involved in the Apoptosis of Human Umbilical Vein Endothelial Cells Induced by Proteasome Inhibitor MG132

作　　者：郭芳[1] 孙文清[1] 屈顺林[1] 何慧[1] 杨向东[2] 

机构地区：[1]南华大学病理生理学教研室,湖南衡阳421001 [2]南华大学心血管病研究所

出　　处：《南华大学学报（医学版）》2009年第2期136-139,共4页Journal of Nanhua University(Medical Edition)

基　　金：湖南省科技厅基金(06FJ3098);湖南省卫生厅基金(C2006-024)资助

摘　　要：目的观察蛋白酶体抑制剂MG132对人血管脐静脉内皮细胞(ECV-304)的致凋亡作用,并从泛素-蛋白酶体途径(UPP)相关基因E1、E2和E3及天冬氨酸特异的半胱氨酸蛋白酶3(Caspase-3)表达初步探讨MG132的致细胞凋亡机制。方法采用蛋白酶体抑制剂MG132(2μmol/L和5μmol/L)处理ECV-304细胞;流式细胞术检测细胞凋亡率;逆转录-聚合酶链反应(RT-PCR)检测UPP相关基因E1、E2、E3和凋亡相关基因caspase-3的转录水平;免疫细胞化学检测细胞caspase-3蛋白表达。结果MG132处理组细胞的凋亡率明显增加;细胞内UPP相关基因E1、E2、E3 mRNA表达下降,而凋亡相关基因caspase-3 mRNA表达上调;细胞内caspase-3蛋白表达水平升高。结论蛋白酶体抑制剂MG132能够诱导ECV-304细胞凋亡,其机制可能是MG132抑制UPP活性,使细胞内caspase-3蛋白降解减少,同时上调caspase-3基因转录而促进细胞凋亡。Objective To study the effect of proteasome inhibitor MG132 on expression of caspase - 3 and UPP associated genes in the apoptosis of human umbilical vein endothelial cells induced. Methods human umbilical vein endothelial cells were treated with MG132 （2 μmol/L, 5 μmol/L） for 24 hours. The apoptotic cells were detected by flow tyrometric analysis. The level of E1, E2, E3 and caspase - 3 mRNA expression were detected by reverse transcription - polymerase chain reaction （ RT - PCR ）. Caspase -3 protein expression was detected by immunocytoehemistry. Results The results showed that E1, E2, E3 genes expression were decreased in treatment group and apoptosis rate were increased obviously. MG132 could up - regulate the gene/protein expression of caspase - 3. Conclusions Protesome inhibitor MG132 could induced human umbilical vein endothelial cells apoptosis by inhibitting UPP activity, up - regulating the gene/protein expression of caspase - 3.

关 键 词：泛素-蛋白酶体途径 蛋白酶体抑制剂 血管内皮细胞 凋亡 CASPASE-3 

分 类 号：R541.4[医药卫生—心血管疾病]