Up-regulation of endothelial nitric oxide synthase by cytochroine P450 arachidonic acid epoxygenase BM3F87V  

Up-regulation of endothelial nitric oxide synthase by cytochrome P450 arachidonic acid epoxygenase BM_3F87V

作　　者：WANG Hong, LIN Li, LU Zaiying and WANG Daowen(Division of Cardiology, Department of Internal Medicine, Tongji Hospital, Tongji Medical College of Huazhong University of Science and Technology, Wuhan 430030, China) 

出　　处：《Progress in Natural Science:Materials International》2002年第8期598-602,共5页自然科学进展·国际材料（英文版）

基　　金：Supported by the National Natural Science Fundation of China (Grant No. 39870307)

摘　　要：Cytochrome P450 (CYP)-dependent metabolites of arachidonic acid, epoxyeicosatrienoic acids (EETs), have been suggested to be an endothelium-derived hyperpolarizing factor (EDHF). However, the interaction or relation between EDHF and endothelial nitric oxide synthase (eNOS) is still to be elucidated. In the present study, the regulation of eNOS by endogenous EDHF is examined. The cytochrome P450 epoxygenase BM3F87V is cloned into the mammalian expression vector pCB6. Cultured bovine aortic endothelial cells (BAECs) less than 4 passages are used and transfected with BM3F87V. The effects of endogenous EETs result from BM3F87V transfection on eNOS are assessed in the endothelial cells by Western blot and Northern blot, and eNOS activity is also measured by the conversion of L-arginine to L-citrulline. Compared to transfection with the empty pCB6 vector, transfection of BAECs with BM3F87V significantly elevates the levels of eNOS protein expression, which is markedly inhibited by treatment with CYP inhibitor 17-ODYA. BM3F87V transfection also elevates the eNOS mRNA level and increases the eNOS activity. This study suggests that EDHF up-regulates eNOS gene expression.Cytochrome P450 (CYP)-dependent metabolites of arachidonic acid, epoxyeicosatrienoic acids (EETs), have been suggested to be an endothelium-derived hyperpolarizing factor (EDHF) . However, the interaction or relation between EDHF and endothelial nitric oxide synthase (eNOS) is still to be elucidated. In the present study, the regulation of eNOS by endogenous EDHF is examined. The cytochrome P450 epoxygenase BM3F87V is cloned into the mammalian expression vector pCB6. Cultured bovine aortic endothelial cells (BAECs) less than 4 passages are used and transfected with BM3F87V. The effects of endogenous EETs result from BM3F87V transfection on eNOS are assessed in the endothelial cells by Western blot and Northern blot, and eNOS activity is also measured by the conversion of L-arginine to L-citrulline. Compared to transfection with the empty pCB6 vector, transfection of BAECs with BM3F87V significantly elevates the levels of eNOS protein expression, which is markedly inhibited by treatment with CYP inhibitor 17-ODYA. BM3F87V transfection also elevates the eNOS mRNA level and increases the eNOS activity. This study suggests that EDHF up-regulates eNOS gene expression.

关 键 词：endothelium-derived hyperpolarizing factor 14 15-epoxyeicosatrienoic acid endothelial nitric oxide synthase en-dothelia] cells BM3F87V. 

分 类 号：R341[医药卫生—基础医学]