摘要
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 possible mechanism of 14,15-epoxyeicosatrienoic acid （14,15-EET） to the palmitate-induced injury of rat cardiomyocyte H9c2 cells. Methods： The H9c2 cells were divided into control group（A）, vehicle group（B） ,palrnitate group（C） ,and palmi- tate＋14,15-EET group（D）. After treatment, H9c2 cells were analyzed by MTT for cell viability, flow cytometry for apoptosis and Western blot for the expression of Bax and Bcl-2. Results： Palmitate significantly decreased the viability of H9c2 cells, the palmitate-induced apoptosis and cell injury were markedly attenuated by 14,15-EET. The inhibition of AKT and ERK1/2 signa- ling inhibited the anti-apoptotic effect of 14,15-EET. Further studies showed that palmitate de- creased expression of the anti-apoptotic protein Bcl-2, increased the expression of pro-apoptotic protein Bax. 14, 15-EET reversed these effects. Inhibition of AKT and ERK1/2 signaling markedly decreased the effect of 14,15-EET on the expression of Bax and Bcl-2 （All P〈0.05）. Conclusion. Palmitate could induce the apoptosis of H9c2 cells, 14,15-EET could inhibitthe ap- optosis of H9c2 cells by down-regulation of Bax and up-regulation of Bcl-2. All these effects were possibly regulated by ERK1/2 and AKT signaling pathways.
出处
《武汉大学学报（医学版）》
CAS
2016年第5期694-698,713,共6页
Medical Journal of Wuhan University
