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

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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。

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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

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 IC50 concentration of TNF-α on human cardiomyocytes cell line HCM, this assays were also performed to analyze the effect of the treatment with IC50 concentration 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 of PAPR-1 gene 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 of PARP-1 gene 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.

Key words： tumor necrosis factor-α myocytes, cardiac PARP-1 inhibitor apoptosis

收稿日期: 2017-01-10

基金资助: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）。

通讯作者: 卢中秋，主任医师，Email：lzq640815@163.com。

作者简介: 谢月群（1985-），女，浙江瑞安人，主治医师，硕士。

链接本文:

https://xb.wmu.edu.cn/CN/ 或 https://xb.wmu.edu.cn/CN/Y2017/V47/I9/654

[1] TIAN M, YUAN Y C, LI J Y, et al. Tumor necrosis factor-a and its role as a mediator in myocardial infarction: A brief review[J]. 慢性疾病与转化医学 (英文), 2015, 1(1): 18-26.
[2] EREMENKO A A, CHERNOVA E V, VINNITSKIĬ L I, et al. Effect of clarithromycin on the systemic inflammatory response syndrome severity in patients after myocardial revascularization surgery[J]. Anesteziol Reanimatol, 2012(3): 67-71.
[3] ADAMY C, LE C P, CANDIANI G, et al. Tumor necrosis factor alpha and glutathione interplay in chronic heart failure[J]. Arch Mal Coeur Vaiss, 2005, 98(9): 906-912.
[4] DE MURCIA J M, NIEDERGANG C, TRUCCO C, et al. Requirement of poly (ADP-ribose) polymerase in recovery from DNA damage in mice and in cells[J]. Proc Natl Acad Sci U S A, 1997, 94(14): 7303-7307.
[5] GIBSON B A, KRAUS W L. New insights into the molecular and cellular functions of poly (ADP-ribose) and PARPs [J]. Nat Rev Mol Cell Biol, 2012, 13(7): 411-424.
[6] SISTIGU A, MANIC G, OBRIST F, et al. Trial Watch-In-hibiting PARP enzymes for anticancer therapy[J]. Mol CellOncol, 2015, 3(2): e1053594.
[7] BERGER N A, SIMS J L, CATINO D M, et al. Poly (ADP-ribose) polymerase mediates the suicide response to massive DNA damage: studies in normal and DNA-repair defective cells[J]. Princess Takamatsu Symp, 1983, 13: 219-226.
[8] KABLAK-ZIEMBICKA A, PRZEWLOCKI T, SOKOŁO-WSKI A, et al. Carotid intima-media thickness, hs-CRP and TNF-α are independently associated with cardiovascular event risk in patients with atherosclerotic occlusive disease [J]. Atherosclerosis, 2011, 214(1): 185-190.
[9] 张利群, 陈冬, 齐国先. 米诺环素后处理通过抑制PARP过度活化减轻心肌缺血/再灌注损伤[J]. 中国病理生理杂志,2015, 31(11): 2009-2015.
[10] YAMAZAKI K, TANAKA S, SAKATA R, et al. Protective effect of cardioplegia with poly (ADP-ribose) polymerase-1 inhibitor against myocardial ischemia-reperfusion injury:in vitro study of isolated rat heart model[J]. J Enzyme Inhib Med Chem, 2013, 28(1): 143-147.
[11] KHAN T A, RUEL M, BIANCHI C, et al. Poly (ADP-ribose) polymerase inhibition improves postischemic myocardial function after cardioplegia-cardiopulmonary bypass[J].J Am Coll Surg, 2003, 197(2): 270-277.
[12] SZABÓ G, BUHMANN V, ANDRÁSI T, et al. Poly-ADP-ribose polymerase inhibition protects against myocardial and endothelial reperfusion injury after hypothermic cardiac arrest[J]. J Thorac Cardiovasc Surg, 2003, 126(3): 651-658.
[13] VUONG B, HOGAN-CANN A D, ALANO C C, et al. NF-κB transcriptional activation by TNFα requires phospholipase C, extracellular signal-regulated kinase 2 and poly (ADP-ribose) polymerase-1[J]. J Neuroinflamm, 2015, 12:229.
[14] MARTINEZ-ZAMUDIO R I, HA H C. PARP1 enhances inflammatory cytokine expression by alteration of promoter chromatin structure in microglia[J]. Brain Behav, 2014, 4(4): 552-565.