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目的探讨普罗布考对过氧化氢诱导的新生大鼠心肌细胞凋亡及肿瘤坏死因子(tumor necrosis factor,TNF)-α生成的影响。方法培养新生大鼠心肌细胞,随机分为A,B,C,D 4组,A组加入0.2 mmol/L过氧化氢;B,C,D组分别给予1,10,100μmol/L普罗布考预处理6 h后,再分别加入0.2 mmol/L过氧化氢继续孵育6 h。采用流式细胞仪检测细胞凋亡;RT-PCR检测心肌细胞TNF-α mRNA、Bcl-2 mRNA及Bax mRNA表达情况;ELISA法检测细胞培养液中TNF-α水平。结果与A组比较,B,C,D组细胞凋亡率均明显降低,Bax mRNA表达下降,Bcl-2 mRNA表达升高,心肌细胞TNF-αmRNA表达及细胞培养液中TNF-α水平均明显降低,差异均有统计学意义(P<0.05)。心肌细胞凋亡率与培养液中TNF-α水平呈正相关(r=0.874,P<0.01)。多元线性回归分析结果表明,凋亡率与培养液中TNF-α水平独立相关(β=0.122,P<0.01)。结论普罗布考抑制过氧化氢诱导的心肌细胞凋亡,其作用机制与抑制TNF-α生成有关。
Objective To investigate the effect of probucol on the cardiomyocyte apoptosis and the production of tumor necrosis factor (TNF) -α induced by hydrogen peroxide in neonatal rats. Methods The neonatal rat cardiomyocytes were cultured and randomly divided into groups A, B, C and D 4, and 0.2 mmol / L hydrogen peroxide was added to group A; and probiotics B, C and D were given respectively to groups 10, After 6 h of treatment, 0.2 mmol / L hydrogen peroxide was added to continue the incubation for 6 h. Cell apoptosis was detected by flow cytometry. TNF-α mRNA, Bcl-2 mRNA and Bax mRNA expression were detected by RT-PCR and TNF-α in cell culture medium by ELISA. Results Compared with group A, the apoptosis rates of B, C and D groups were significantly decreased, the expression of Bax mRNA and Bcl-2 mRNA were increased, the expression of TNF-α mRNA and the level of TNF-α in cell culture medium were significantly increased Reduce, the difference was statistically significant (P <0.05). The apoptosis rate of cardiomyocytes was positively correlated with the level of TNF-α in the culture medium (r = 0.874, P <0.01). The results of multivariate linear regression analysis showed that the apoptosis rate was independent of the level of TNF-αin culture medium (β = 0.122, P <0.01). Conclusions Probucol inhibits hydrogen peroxide-induced cardiomyocyte apoptosis and its mechanism of action is related to the inhibition of TNF-α production.