细胞色素P450表氧化酶经线粒体途径抗内皮细胞凋亡的机制研究

The protect effect of arachidonic acids cytochrome P450 expoxygenases against endothelial cells apoptosis through mitochondrial pathway

目的:研究细胞色素P450表氧化酶对肿瘤坏死因子诱导的内皮细胞抗凋亡的机制。方法:原代培养主动脉内皮细胞(BAECs)2周后,分别转染细胞色素P450表氧化酶基因rAAV-F87V。用肿瘤坏死因子(TNF-α)和放线菌素D(ActD)诱导凋亡,用Western blot法分别检测细胞胞质及线粒体的细胞色素C的含量。以ELISA方法检测Caspase-3、-8、-9的活性。结果:Western Blot结果表明,转染CYP P450表氧化酶后细胞胞质的细胞色素C明显减少,线粒体的细胞色素C明显增加。ELISA结果显示Caspase-3、-8、-9的活性均较对照组明显降低(均P<0.05)。结论:细胞色素P450表氧化酶基因rAAV-F87V抑制内皮细胞凋亡是通过抑制线粒体途径发挥作用的。

Objective：Aim To investigate the protect effect of arachidonic acids cytochrome P450 expoxygenases（CYP450）against endothelial cells apoptosis induced by tumor necrosis factor-α.Method：Arachidonic acids cytochrome P450 expoxygenases（CYP450）genes CYPF87 V,which had been constructed in recombinant adeno-associated viral expression vectors（rAAV）,were transfected into bovine aortic endothelial cells（BAECs）.Two weeks later,BAECs were treated with TNF-αand ActD.The level of cytochrome C（CytC）in cytosol or in mitochondria were detected by Western blot.The activity of Casapase-3、-8、-9 were detected by ELISA.Result：CYPF87 elevated Cyt C content in mitochondria and reduced Cyt C content in cytoplasm.Furthermore,CYPF87 Vdecreased the activation of caspase-3,-8 and-9.Conclusion：Epoxygenase reduced the apoptosis of endothelial cells through mitochondrial pathway.