MG132诱导人血管平滑肌细胞凋亡及对caspase3表达的影响  

Effect on expression of caspase-3 in the apoptosis of cultured human umbilical Vascular Smooth Muscle cells induced by Proteasome Inhibitor MG132

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

机构地区：[1]南华大学病理生理学教研室,衡阳421001 [2]南华大学分子生物中心

出　　处：《中国药物与临床》2009年第5期381-383,450,共4页Chinese Remedies & Clinics

基　　金：湖南省科技厅基金(06FJ3098)

摘　　要：目的观察蛋白酶体抑制剂MG132对人脐静脉血管平滑肌细胞(VSMC)的致凋亡作用及其对凋亡相关的天冬氨酸特异的半胱氨酸蛋白酶3(caspase3)表达的影响。方法采用多个浓度(2.5,5,10μmol/L)的蛋白酶体抑制剂MG132处理人脐静脉VSMC48h;DNA琼脂糖凝胶电泳检测细胞凋亡;流式细胞术分析细胞周期和细胞凋亡率;反转录-聚合酶链反应(RT-PCR)检测caspase3基因转录水平;免疫细胞化学检测细胞caspase3蛋白表达。结果蛋白酶体抑制剂MG132处理48h后细胞凋亡率增加;RT-PCR检测发现细胞内凋亡相关基因caspase3mRNA表达上调;免疫细胞化学检测细胞caspase3蛋白表达水平升高。结论蛋白酶体抑制剂MG132能够诱导人脐静脉VSMC凋亡,其作用呈量-效关系;MG132诱导VSMC凋亡的机制可能与MG132抑制泛素-蛋白酶体途径(UPP)活性,促进caspase3基因转录,使细胞内caspase3增加而促进细胞凋亡。Objective To study the effect of proteasome inhibitor MG132 on expression of caspase3 and apoptosis in cultured human umbilical vascular smooth muscle cells. Methods human umbilical vascular smooth muscle cells were treated with MG132 （2.5μmol·L^-1, 5μmol·L^-1, 10μmol·L^-1） for 48 hours.The apoptotic cells were determined by DNA fragment analysis and flow cytometric analysis. The level of caspase3 mRNA expression were detected by reverse transcription-polymerase chain reaction （RT-PCR）. Caspase3 protein expression was detected by immunocytochemistry. Results The rate of apoptosis as increased obviously after treatment with MG132 for 48 hours. RTPCR showed up-regulated gene/protein expression of caspase3. Conclusions Proteasome inhibitor MG132 could induce human umbilical vascular smooth muscle cells in a dose-dependent manner; Possibly through MG132 inhibition on upp activity which promotes caspase3 gene transcription and thereby leads to increased production of intracellular caspase3 protein.

关 键 词：肌 平滑 血管 细胞凋亡 泛素蛋白连接酶复合物 天冬氨酸特异的半胱氨酸蛋白酶3 

分 类 号：R96[医药卫生—药理学]