PARP-1蛋白抑制剂降低TNF-α介导的人心肌细胞生长抑制和凋亡  

PARP-1 protein inhibitor attenuated the growth inhibition and apoptosis of human cardiomyocytes induced by TNF-α

作　　者：谢月群 王蕾[1] 陈玲珑[1] 徐颖 杨之涛[2] 卢中秋[3] XIE Yuequn;WANG Lei;CHEN Linglong;XU Ying;YANG Zhitao;LU Zhongqiu(Department of Emergency, Wenzhou People’s Hospital, the Third Clinical Institute Affiliated to Wenzhou Medical University, Wenzhou, 325000;Department of Emergency, Ruijin Hospital Shanghai JiaoTong University School of Medicine, Shanghai, 200025;Department of Emergency, the First Affiliated Hospital of Wenzhou Medical University, Wenzhou, 325015)

机构地区：[1]温州医科大学第三临床学院温州市人民医院急诊科,浙江温州325000 [2]上海交通大学医学院附属瑞金医院急诊科,上海200025 [3]温州医科大学附属第一医院急诊医学中心,浙江温州325015

出　　处：《温州医科大学学报》2017年第9期654-659,共6页Journal of Wenzhou Medical University

基　　金：浙江省医学创新学科建设计划项目(11-CX26);浙江省中医药重点学科计划项目(2012-XK-A28);浙江省"十二五"重点学科建设项目(2012-207);温州市科技局科研基金资助项目(Y20170736)

摘　　要：目的:研究PARP-1蛋白抑制剂对由肿瘤坏死因子α(TNF-α)介导的人心肌细胞生长抑制和凋亡的干预作用。方法:MTT比色法检测TNF-α抑制人心肌细胞HCM细胞株生长的IC_(50)浓度,以及PARP-1蛋白抑制剂干预对HCM细胞生长抑制率的影响;流式细胞术检测TNF-α和PARP-1蛋白抑制剂对HCM细胞的细胞凋亡比率的影响;RT-PCR和Western blot分析PARP-1蛋白抑制剂干预对PARP-1基因的mRNA和蛋白水平表达影响。结果:TNF-α对HCM细胞具有明显细胞增殖抑制和细胞凋亡诱导作用,并且上调PARP-1基因mRNA水平,促进PARP-1蛋白裂解(P<0.05);PARP-1蛋白抑制剂干预后,TNF-α对HCM细胞的生长抑制和诱导细胞凋亡作用均减弱(P<0.05),PARP-1基因表达下调(P<0.05),PARP-1蛋白和其裂解产物与对照组比较差异无统计学意义(P>0.05)。结论:通过PARP-1蛋白抑制剂阻断PARP-1蛋白活性和基因的转录水平可以减弱TNF-α对HCM的细胞生长抑制和细胞凋亡诱导作用。Objective:To study the effect of PARP-1inhibitor(4-Aminonaphthalimide)on the apoptosisof human cardiomyocytes mediated by TNF-α.Methods:MTT assays were used to detect the IC50concentrationof TNF-αon human cardiomyocytes cell line HCM,this assays were also performed to analyze the effect of thetreatment with IC50concentration of TNF-αcombined with different concentration of PARP-1inhibitor on proliferationof the HCM cells.Flow cytometry was used to monitor the apoptosis of HCM cells treated with differentconcentrations of TNF-αwith or without PARP-1inhibitors;RT-PCR and Western blot were used to analyze theexpression level of PARP-1in HCM cells after treated with different concentration of TNF-αwith or withoutPARP-1inhibitor.Results:TNF-αcould perform the inhibitory effect of proliferation and induced the apoptosisof HCM cells,TNF-αalso induced the degradation of PARP-1protein and up-regulated expression of PAPR-1gene in HCM cells(P<0.05);The inhibit effect of proliferation and the apoptosis rate of HCM cells inducedby TNF-αwas decreased after the intervention of PARP-1inhibitors(P<0.05).The mRNA expression level ofPARP-1gene was down-regulated and the difference between the intervention group and the control group wasnot significantly in protein level.Conclusion:The effect of cells growth inhibition and apoptosis induced byTNF-αon HCM can be attenuated by blocking the activity of PARP-1protein and gene transcription.

关 键 词：肿瘤坏死因子Α 肌细胞 心脏 PARP-1蛋白抑制剂 细胞凋亡 

分 类 号：R541[医药卫生—心血管疾病]