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的影响  

EFFECT OF CYTOCHROME P450 EXPOXYGENASES ON VESSEL ENDOTHELIAL CELL APOPTOSIS INDUCED BY TUMOR NECROSIS FACTOR ALPHA

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作  者:陈积雄[1] 蒋建刚[1] 汪道文[1] 

机构地区:[1]华中科技大学同济医学院附属同济医院心血管内科研究室武汉,430030

出  处:《中国组织化学与细胞化学杂志》2005年第4期361-365,共5页Chinese Journal of Histochemistry and Cytochemistry

基  金:国家自然基金资助课题(30270561)

摘  要: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(bovineaorticendothelialcells,BAECs)凋亡的影响。方法在原代培养的BAECs中,分别转染构建在腺相关病毒载体内的细胞色素P450表氧化酶基因2J2,2C11,F87V两周后,用肿瘤坏死因子(TNF-α)(10ng/ml)和放线菌素D(ActD)(5ng/ml)诱导内皮细胞凋亡,通过细胞形态学和流式细胞计数观察其凋亡变化,同时采用Westernblot分析检测Bcl-2和P38和丝裂原激动蛋白激酶ERK的表达水平。结果与对照相比,转染细胞色素P450表氧化酶基因后BAECs凋亡细胞数减少,Bcl-2表达增加,P38表达减低,ERK的磷酸化水平增加。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BAECs凋亡,并可能通过上调Bcl-2、下调P38的表达水平和通过增加ERK的磷酸化水平发挥抗凋亡作用。Objective To observe the effect of cytochrome P450 expoxygenases (CYP450) on the apoptosis of primarily cultured bovine aortic endothelial cells (BAECs) induced by tumor necrosis factor-α. Methods Arachidonic acide CYP450 genes-CYP2J2, CYPF87V and CYP2Cll-which had been constructed in recombinant adeno-associated viral expression vectors (rAAV), were transfected into cultured BAECs. Two weeks later, BAECs were treated with TNF-α (10ng/ml) and ActD (5ng/ml). Apoptosis was checked by morphological observation and flow-cytometry assay. The levels of Bcl-2, P38 and phospho-ERK expression in the cells were determined by Western blot. Results Compared with the controls, the CYP450 genes significantly inhibited TNF-α-induced BAEC apoptosis. Furthermore, the expression of CYP450 genes significantly up-regulated Bcl-2 protein expression, down-regulated P38 expresssion and increased ERK phosphorylation. Conclusion The results indicate that CYP450 can protect vessel endothelial cells from apoptosis induced by TNF-α and the antiapoptotic effect of CYP450 mediated by up-regulating Bcl-2, increasing the phosphorylation level of ERK and down-regulating P38 protein expression.

关 键 词:细胞色素表氧化酶 肿瘤坏死因子 内皮细胞 凋亡 

分 类 号:R541[医药卫生—心血管疾病]

 

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