PARP-1蛋白抑制劑降低TNF-α介導的人心肌細胞生長抑制和凋亡

謝月群，王蕾，陳玲瓏，徐穎，楊之濤，盧中秋

（1.溫州醫科大學第三臨床學院 溫州市人民醫院 急診科，浙江 溫州 325000；2.上海交通大學醫學院附屬瑞金醫院 急診科，上海 200025；3.溫州醫科大學附屬第一醫院 急診醫學中心，浙江 溫州325015）

PARP-1蛋白抑制劑降低TNF-α介導的人心肌細胞生長抑制和凋亡

謝月群1，王蕾1，陳玲瓏1，徐穎1，楊之濤2，盧中秋3

（1.溫州醫科大學第三臨床學院 溫州市人民醫院 急診科，浙江 溫州 325000；2.上海交通大學醫學院附屬瑞金醫院 急診科，上海 200025；3.溫州醫科大學附屬第一醫院 急診醫學中心，浙江 溫州325015）

目的：研究PARP-1蛋白抑制劑對由腫瘤壞死因子α（TNF-α）介導的人心肌細胞生長抑制和凋亡的干預作用。方法：MTT比色法檢測TNF-α抑制人心肌細胞HCM細胞株生長的IC50濃度，以及PARP-1蛋白抑制劑干預對HCM細胞生長抑制率的影響；流式細胞術檢測TNF-α和PARP-1蛋白抑制劑對HCM細胞的細胞凋亡比率的影響；RT-PCR和Western blot分析PARP-1蛋白抑制劑干預對PARP-1基因的mRNA和蛋白水平表達影響。結果：TNF-α對HCM細胞具有明顯細胞增殖抑制和細胞凋亡誘導作用，并且上調PARP-1基因mRNA水平，促進PARP-1蛋白裂解（P＜0.05）；PARP-1蛋白抑制劑干預后，TNF-α對HCM細胞的生長抑制和誘導細胞凋亡作用均減弱（P＜0.05），PARP-1基因表達下調（P＜0.05），PARP-1蛋白和其裂解產物與對照組比較差異無統計學意義（P＞0.05）。結論：通過PARP-1蛋白抑制劑阻斷PARP-1蛋白活性和基因的轉錄水平可以減弱TNF-α對HCM的細胞生長抑制和細胞凋亡誘導作用。

腫瘤壞死因子α；肌細胞，心臟；PARP-1蛋白抑制劑；細胞凋亡

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.

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

腫瘤壞死因子-α（tumor necrosis factor-α，TNF-α）作為一種關鍵的調節因子參與炎癥反應，除了通過與其受體結合啟動炎癥免疫應答，還可以作為獨立因素參與心肌損傷[1]。……