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目的:观察芝麻素含药血清对血管紧张素Ⅱ(AngⅡ)诱导大鼠心肌细胞凋亡的影响,并探讨其可能机制。方法:培养H9C2大鼠心肌细胞,与芝麻素含药血清或空白血清预孵12 h后,加入AngⅡ孵育24 h。MTT法检测心肌细胞活力,流式细胞术检测细胞凋亡率,Western blot法检测BCL-2、BAX、Caspase-3、SOD和p47phox蛋白表达水平,比色法测定细胞总抗氧化能力、细胞内活性氧(ROS)水平和丙二醛(MDA)含量。结果:芝麻素含药血清预孵可显著改善AngⅡ诱导的心肌细胞凋亡和活力损伤(P<0.05或P<0.01),下调BAX、Caspase-3及p47phox蛋白表达,上调BCL-2、SOD蛋白表达水平(P<0.05或P<0.01)。此外,芝麻素含药血清可显著提高心肌细胞总抗氧化能力降低细胞内ROS和培养上清MDA含量(P<0.05或P<0.01)。空白血清对上述指标均无显著影响。结论:芝麻素可显著抑制AngⅡ诱导的心肌细胞凋亡和活力损伤,其机制与改善氧化应激状态、调节BCL-2/BAX蛋白表达水平,进而下调Caspase-3蛋白表达有关。
Objective: To investigate the effects of sesamin-containing serum on cardiomyocyte apoptosis induced by angiotensin Ⅱ (AngⅡ) in rats and its possible mechanism. Methods: H9C2 rat cardiomyocytes were incubated with sesamin-containing serum or blank serum for 12 h, then incubated with AngⅡ for 24 h. The viability of cardiomyocytes was measured by MTT assay. The apoptosis rate was detected by flow cytometry. The protein expressions of BCL-2, BAX, Caspase-3, SOD and p47phox were detected by Western blot. The total antioxidant capacity, Reactive oxygen species (ROS) levels and malondialdehyde (MDA) content. Results: Preincubation with sesamin-containing serum significantly (P <0.05 or P <0.01) decreased AngⅡ-induced cardiomyocyte apoptosis and down-regulated the protein expression of BAX, Caspase-3 and p47phox, Expression level (P <0.05 or P <0.01). In addition, sesamin-containing serum could significantly increase the total anti-oxidative ability of cardiomyocytes and decrease the content of intracellular ROS and the content of MDA in cultured supernatant (P <0.05 or P <0.01). Blank serum had no significant effect on the above indicators. Conclusion: Sesamin can significantly inhibit Ang Ⅱ-induced cardiomyocyte apoptosis and its activation. Its mechanism is related to the improvement of oxidative stress, regulation of BCL-2 / BAX protein expression and down-regulation of Caspase-3 protein expression.