利用代谢组学研究血流剪切力对内皮细胞类二十烷酸代谢产物的影响

Effect of shear stress on eicosanoid metabolism in endothelial cells by metabolomic approach

本文旨在研究血流剪切力对内皮细胞多不饱和脂肪酸产生的类二十烷酸代谢产物的影响,以阐明不同模式的血流剪切力对内皮细胞功能的影响及机制。首先,分别给予人脐静脉内皮细胞对照静止(Static)、层流型剪切力(laminar shear stress,LSS)、振荡型剪切力(oscillatory shear stress,OSS)处理6 h后,用新鲜的M199培养液孵育内皮细胞3 h,利用超高效液相色谱-质谱联用技术检测孵育培养液中内皮细胞分泌的类二十烷酸代谢产物的水平。结果显示:在不同的血流剪切力作用下,类二十烷酸代谢的水平有显著改变。我们筛选出OSS组相对于LSS组显著上调的10种代谢物,包括由环氧合酶产生的代谢产物PGD2、PGE2、PGF2α以及PGJ2,由脂氧合酶或自发氧化作用产生的11-HETE、15-HETE、13-HDoHE,以及由细胞色素P450氧化酶和可溶性环氧化物水解酶产生的12,13-EpOME、9,10-EpOME、9,10-DiHOME,并从离体和在体水平验证了类二十烷酸代谢酶mRNA表达水平升高。以上结果表明OSS可能通过上调代谢酶基因的转录水平促进类二十烷酸代谢产物的升高,从而在动脉粥样硬化发生与发展中起关键作用。本研究利用代谢组学技术从整体上研究了类二十烷酸代谢产物对血流剪切力的响应,为血流动力学的系统生物学研究做出了重要的补充。

The article aims to study the effect and mechanism of shear stress on eicosanoids produced by the metabolism of polyunsaturated fatty acids in endothelial cells.First,human umbilical vein endothelial cells were treated by control(Static),laminar shear stress(LSS)and oscillatory shear stress(OSS)for 6 h.Then the endothelial cells were incubated with fresh M199 medium for 3 h,and the cell culture medium was collected.Ultra-performance liquid chromatography-mass spectrometer was used to detect the level of eicosanoid metabolites secreted by endothelial cells.The results showed that under different shear stress,the level of eicosanoid metabolites were changed significantly.We found 10 metabolites were significantly up-regulated by OSS compared with those in LSS group,including PGD2,PGE2,PGF2αand PGJ2 produced by cyclooxygenase;11-HETE,15-HETE,13-HDoHE produced by lipoxygenase or spontaneous oxidation;12,13-EpOME,9,10-EpOME,9,10-DiHOME produced by cytochrome P450 oxidase and soluble epoxide hydrolase.The transcription levels of these up-regulated eicosanoids metabolic enzyme-related genes were also increased in vitro and in vivo.These results indicate that OSS may promote the increase of metabolites by up-regulating the transcription level of metabolic enzyme-related genes,which playing a key role in the development of atherosclerosis.This study reveals the effect of shear stress on eicosanoid metabolism in endothelial cells,which provides a novel supplement to the systems biology approach to study systemic hemodynamics.