维生素C对乌头碱中毒导致乳鼠心肌细胞凋亡的影响及机制分析

Effect and mechanism of vitamin C on cardiomyocyte apoptosis induced by aconitine

目的观察维生素C对乌头碱导致心肌细胞损伤的防护作用并探讨可能的机制。方法提取并培养乳鼠原代心肌细胞,采用蛋白质印迹法验证浓度梯度的乌头碱对心肌细胞凋亡的影响;加入浓度梯度的维生素C,用同样的实验方法观察其对乌头碱中毒导致心肌细胞凋亡的改善作用。利用蛋白质印迹法筛选出在整个中毒解救过程中p53蛋白表达的变化;探讨维生素C联合应用p53抗体对乌头碱导致心肌细胞凋亡的影响。结果乌头碱能够浓度梯度性地促进Bad蛋白表达并下调Bcl-2蛋白表达(P<0.01),说明其能诱导心肌细胞凋亡。维生素C可以减弱乌头碱中毒心肌细胞中Bad蛋白的表达而增强Bcl-2的表达(P <0.05),说明其能减弱乌头碱中毒引起的细胞凋亡。浓度梯度的维生素C与乌头碱合用时,p53蛋白的表达随着维生素C浓度的增加而降低(P <0. 01)。加入外源性p53抗体,可以减弱乌头碱中毒引起的细胞凋亡,增强维生素C的解救作用(P<0.01)。结论维生素C可以减轻乌头碱的毒性作用,这可能与维生素C抑制了p53信号通路有关。

Objective To investigate the effect of vitamin C on cardiomyocyte apoptosis caused by aconitine and to analyze the potential mechanism. Methods Primary cardiomyocytes of suckling mice were extracted and cultured. Western blotting was used to detect the effects of aconitine and vitamin C on cell growth and the effect of vitamin C combined with p53 on cell apoptosis induced by aconitine. Results Aconitine could increase Bad expression and reduce Bcl-2 expression(P<0.01), suggesting it induced the apoptosis of cardiomyocytes. Vitamin C could reduce Bad expression and increase Bcl-2 expression in cardiomyocytes cultured with aconitine(P<0.05);it could alleviate the toxicity of aconitine. Under gradient concentrations of vitamin C plus aconitine, the expression level of p53 in cells decreased with the concentration of vitamin C(P<0.01). P53 anti-body could reduce the toxicity of aconitine and enhance the effect of vitamin C(P<0.01). Conclusion Vitamin C can alleviate the toxicity of aconitine on cardiomyocytes, which may be mediated by the p53 pathway.