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
诱导,1材料和方法,2结果,3讨论
谢月群,王蕾,陈玲珑,徐颖,杨之涛,卢中秋(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)
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
谢月群1,王蕾1,陈玲珑1,徐颖1,杨之涛2,卢中秋3
(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)
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IC50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。
肿瘤坏死因子α;肌细胞,心脏;PARP-1蛋白抑制剂;细胞凋亡
Abstract: Objective:To study the effect of PARP-1 inhibitor (4-Aminonaphthalimide) on the apoptosis of human cardiomyocytes mediated by TNF-α.Methods:MTT assays were used to detect the IC50concentration of TNF-α on human cardiomyocytes cell line HCM, this assays were also performed to analyze the effect of the treatment with IC50concentration of TNF-α combined with different concentration of PARP-1 inhibitor on proliferation of the HCM cells. Flow cytometry was used to monitor the apoptosis of HCM cells treated with different concentrations of TNF-α with or without PARP-1 inhibitors; RT-PCR and Western blot were used to analyze the expression level of PARP-1 in HCM cells after treated with different concentration of TNF-α with or without PARP-1 inhibitor.Results:TNF-α could perform the inhibitory effect of proliferation and induced the apoptosis of HCM cells, TNF-α also induced the degradation of PARP-1 protein and up-regulated expression ofPAPR-1gene in HCM cells (P<0.05); The inhibit effect of proliferation and the apoptosis rate of HCM cells induced by TNF-α was decreased after the intervention of PARP-1 inhibitors (P<0.05). The mRNA expression level ofPARP-1gene was down-regulated and the difference between the intervention group and the control group was not significantly in protein level.Conclusion:The effect of cells growth inhibition and apoptosis induced by TNF-α on HCM can be attenuated by blocking the activity of PARP-1 protein and gene transcription. ......
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