TRAIL介导SEB-1皮脂细胞凋亡中MG-132的促进机制研究  

The mechanism ofMG-132 on promotingTRAIL-mediated SEB-1 sebocyte apoptosis

作　　者：柳林[1] 苏江维[1] Liu Lin;Su Jiangwei(Department of Dermatology,The Central Hospital of E’zhou City,E’Zhou 436000,China)

机构地区：[1]湖北省鄂州市中心医院皮肤科,鄂州436000

出　　处：《广西医科大学学报》2020年第12期2186-2191,共6页Journal of Guangxi Medical University

基　　金：湖北省科技厅科研计划项目(No.2019YFS0436)。

摘　　要：目的:研究肿瘤坏死因子相关的凋亡诱导配体(TRAIL)介导SEB-1皮脂细胞凋亡中MG-132的促进机制。方法:将实验SEB-1皮脂细胞随机分为正常组、MG-132组及siRNA+MG-132组。正常组正常培养传代,MG-132组加入蛋白酶体抑制剂MG-132,siRNA+MG-132组在干扰处理TRAIL基因以后加入蛋白酶体抑制剂MG-132。采用MTT法检测细胞凋亡率。分别使用免疫印迹法(Western blotting)及实时荧光定量PCR(qPCR)法对蛋白及基因表达量进行检测。结果:24 h时正常组和MG-132组的细胞数量明显高于12 h,且正常组较高(P<0.05)。12 h至24 h内,siRNA+MG-132组细胞的细胞数量较MG-132组升高幅度更大(P<0.05);随着培养时间的延长,MG-132组细胞的凋亡率逐渐升高,且高于正常组(P<0.05);相比于MG-132组,siRNA+MG-132组培养12 h和24 h后细胞凋亡率则明显降低(P<0.05)。与正常组相比,MG-132组及siRNA+MG-132组细胞Bik、Bid基因表达量明显升高(P<0.05),siRNA+MG-132组的Bik、Bid凋亡基因表达量低于MG-132组(P<0.05);MG-132组及siRNA+MG-132组细胞的Caspase8、TRAIL和Bax蛋白表达量明显升高,Bcl-2蛋白表达量明显降低(P<0.05),siRNA+MG-132组细胞蛋白表达量变化明显低于MG-132组(P<0.05)。结论:蛋白酶体抑制剂MG-132对TRAIL介导的SEB-1皮脂细胞凋亡有促进作用。Objective:To study the mechanism of MG-132 on promoting TNF-related apoptosis-inducing ligand(TRAIL)-mediated SEB-1 sebocytes apoptosis.Methods:The SEB-1 cells were divided into 3 groups:normal group,MG-132 group and siRNA+MG-132 group.The normal group cells were normally cultured and passaged.The MG-132 group added proteasome inhibitor MG-132,and siRNA+MG-132 group added MG-132after interfering with the TRAIL gene.MTT method was used to detect the apoptotic rate.Real-time fluorescence quantitative PCR(qPCR)and Western blotting were used to determine the mRNA and protein expressions,respectively.Results:The number of cells in the normal group and the MG-132 group was significantly higher at 24 h than that at 12h,and the normal group was higher(P<0.05).From 12h to 24h,the increase of cells number in the siRNA+MG-132 group was more significant than that in the MG-132 group(P<0.05).With the cultured time prolonged,the apoptotic rate of cells in the MG-132 group increased gradually,and it was higher than that in the normal group(P<0.05).Compared with the normal group,the apoptotic rate of cells in MG-132 group andsiRNA+MG-132 group was significantly reduced after culturing for 12 h and 24 h.The mRNA levels of Bik and Bid in siRNA+MG-132 group were lower than those in MG-132 group(P<0.05).The protein expressions of Caspase8,TRAIL and Baxin MG-132 group and siRNA+MG-132group were significantly increased,while the expression of Bcl-2 protein was decreased,and the changes of protein expressions in siRNA+MG-132 group were more obvious(P<0.05).Conclusion:The proteasome inhibitor MG-132 can promote SEB-1 cell apoptosis mediated by TRAIL.

关 键 词：SEB-1细胞 凋亡 肿瘤坏死因子相关的凋亡诱导配体 MG-132 

分 类 号：R285[医药卫生—中药学]