细胞色素P450表氧化酶对内皮细胞凋亡及NF-κB表达的影响

Effect of cytochrome p450 expoxygenases on vessel endothelial cell apoptosis and the expression of nuclear factor-κB

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凋亡及NF-κB活性的影响。方法在原代培养的牛主动脉内皮细胞中,分别转染细胞色素P450表氧化酶基因rAAV-2J2,rAAV-2C11,rAAV-F87V两周后,用肿瘤坏死因子(TNF-α)(10ng/ml)和放线菌素D(ActD) (5ng/ml)诱导凋亡,通过DNA ladder观察细胞凋亡,同时ELISA法测定NF-κB的活性。结果与对照组相比,转染细胞色素P450表氧化酶基因后能抑制内皮细胞的凋亡,同时诱导细胞凋亡后,转染CYP450各基因组NF-κB中P65的活性较对照组明显降低。结论细胞色素P450表氧化酶能明显的抑制肿瘤坏死因子诱导的内皮细胞凋亡,并通过抑制NF-κB的核转位发挥抗凋亡作用。

Objective To observe the effect of arachidonic acids cytochrome P450 expoxygenases （CYP450） on the apoptosis and activity of nuclear factor-κB （NF-κB） of vessel endothelial cells induced by tumor necrosis factor in primarily cultured bovine aortic endothelial cells （BAECs）. Method arachidonic acids cytochrome P450 expoxygenases（CYP450） genes were transfected by CYP2J2, CYPF87V , CYP2Cll using recombinant adeno-associated viral vector（rAAV） into cultured BAECs for 2 weeks, BAECs were treated with TNF-cα（ 10 ng/ml）and ActD（5 ng/ml）. Apoptotic responses were assessed by DAN ladder and activity of nuclear factor-κB （NF-κB） was tested by ELISA. Result Apoptosis in BAECs was significantly inhibited and the activity of p65 of nuclear factor-κB （NF-κB） was deseased after infected with these CYP450 genes compared with control. Conclusion The results indicated that CYP450 could significantly inhibit the apoptosis in BAECs induced by TNF-αand the antiapoptotic effect were mediated by inhibiting the nuclear translocation of NF-κB.