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目的:观察14,15-环氧化二十碳三烯酸(14,15-EET)对棕榈酸盐(Palmitate)诱导的H9c2大鼠心肌细胞凋亡的影响及其机制。方法:传代培养大鼠心肌细胞H9c2,分为正常组、溶媒组、Palmitate组、Palmitate+14,15-EET组,噻唑蓝比色法(MTT)检测细胞存活率、流式细胞仪检测细胞凋亡以及Western blot检测Bax,Bcl-2蛋白表达。结果:Palmitate的刺激可显著降低H9c2细胞的存活率并呈剂量依赖效应,14,15-EET呈剂量依赖性增加不同浓度Palmitate刺激后H9c2细胞存活率;Palmitate可诱导H9c2细胞凋亡,14,15-EET的作用显著抑制了Palmitate诱导的H9c2细胞的凋亡,联用胞外信号调节蛋白激酶(ERK1/2)及蛋白激酶B(PKB或AKT)信号通路抑制剂显著抑制了14,15-EET的抗凋亡作用。Palmitate的诱导使Bax表达增加,Bcl-2表达减少;14,15-EET可下调Bax,上调Bcl-2的表达,联用ERK1/2及AKT信号通路抑制剂能显著抑制14,15-EET对Palmitate诱导后H9c2细胞Bax表达水平的下调及Bcl-2表达水平上调的作用(P均<0.05)。结论:Plamitate可诱导H9c2细胞凋亡,14,15-EET可通过下调Bax,上调Bcl-2的表达抑制Palmitate诱导的H9c2心肌细胞凋亡作用。这些作用可由ERK1/2和AKT信号通路介导。
Objective: To investigate the effect and mechanism of 14,15-epoxyeicosatrienoic acid (14,15-EET) on cardiomyocyte apoptosis induced by Palmitate in H9c2 rats. Methods: H9c2 cells were subcultured and divided into normal group, vehicle group, Palmitate group, Palmitate + 14,15-EET group and MTT assay. Cell viability was measured by flow cytometry The expression of Bax and Bcl-2 protein were detected by Western blot. Results: Palmitate could significantly reduce the survival rate of H9c2 cells in a dose-dependent manner. 14,15-EET increased the survival rate of H9c2 cells stimulated with different concentrations of Palmitate in a dose-dependent manner. Palmitate induced the apoptosis of H9c2 cells, -EET significantly inhibited the apoptosis of H9c2 cells induced by Palmitate. The combination of ERK1 / 2 and PKB or AKT signaling inhibitors significantly inhibited the 14,15-EET Anti-apoptotic effect. Palmitate induces the increase of Bax expression and the decrease of Bcl-2 expression; 14,15-EET can down-regulate Bax and up-regulate the expression of Bcl-2, and the combination of ERK1 / 2 and AKT signaling pathway inhibitors can significantly inhibit 14,15-EET The expression of Bax and the up-regulation of Bcl-2 in H9c2 cells induced by Palmitate (all P <0.05). Conclusion: Plamitate can induce H9c2 cell apoptosis. 14,15-EET can inhibit the apoptosis of H9c2 cardiomyocytes induced by Palmitate by down-regulating Bax and up-regulating the expression of Bcl-2. These effects are mediated by the ERK1 / 2 and AKT signaling pathways.