11,12-环氧二十碳三烯酸对缺氧/复氧诱导的内皮细胞损伤及细胞间黏附因子-1表达的影响  

Effect of 11,12-Epoxyeicosatrienoic Acids on Injury of Endothelial Cells induced by Hypoxia/reoxygenation and ICAM-1

作　　者：邱笑违[1] 王珏[1] 王红霞[1] 王雯[1] 蒋东桥[1] 芦玲巧[1] 唐朝枢[1] 张立克[1] 

机构地区：[1]首都医科大学基础医学院病理生理学教研室

出　　处：《首都医科大学学报》2009年第6期805-808,共4页Journal of Capital Medical University

基　　金：北京市自然科学基金(7062007)资助项目~~

摘　　要：目的观察11,12-环氧二十碳三烯酸(11,12-epoxyeicosatrienoic acids,11,12-EET)对缺氧/复氧诱导的内皮细胞损伤以及黏附分子表达的影响,了解环氧二十碳三烯酸发挥血管保护作用的可能途径,初步探讨其作用机制。方法采用原代培养人脐静脉内皮细胞,用数字表法将其随机分为对照组、11,12-EET对照组、缺氧/复氧组和11,12-EET缺氧/复氧组。通过向培养瓶内通入混合气体(2%O2,5%CO2,93%N2)3h,复氧1h,复制缺氧/复氧损伤模型。采用MTT法检测细胞活力,用比色法检测培养液中超氧化物歧化酶(SOD)及丙二醛(MDA)的变化,用RT-PCR法检测细胞间黏附分子-1(intercellular adhesion molecule-1,ICAM-1)mRNA表达,用酶联免疫吸附实验测定细胞间黏附分子-1蛋白表达,用Western blotting方法检测蛋白激酶B(Akt)。结果11,12-EET在常氧条件下可对细胞造成轻度损伤,在缺氧/复氧条件下能显著提高内皮细胞的存活率,提高SOD的活性,降低MDA的含量,抑制细胞间黏附分子-1 mRNA和蛋白的表达,提高Akt的表达。结论11,12-EET具有减轻内皮细胞缺氧/复氧损伤作用,这可能与其能提高缺氧/复氧条件下内皮细胞SOD活性、清除氧自由基、抑制细胞间黏附分子-1 mRNA和蛋白的表达和促进Akt的表达有关。Objective To investigate the effects of 11,12-epoxyeicosatrienoic acids（11,12-EET） on injury of endothelial cells induced by hypoxia/reoxygenation and intercellular adhesion molecule-1（ICAM-1）,and reveal the possible pathway of EETs on protection. Methods Primary cultured HUVECs were randomly divided into control group,11,12-EET control group,hypoxia/reoxygenation group,11,12-EET hypoxia/reoxygenation group. Hypoxia/reoxygenation injury model in HUVECs was produced by exposed to hypoxia（2% O2,5% CO2 and 93% N2） for 3 hours,respectively,followed by reoxygenation（95% air and 5% CO2）. The cell viability were monitored by MTT assay. Colorimetry method was used to assay methyl lnedioxyamphetamine（MDA） and activity of superoxide dismutase（SOD） in culture medium. The ICAM-1 mRNA expression was determined by reverse transcriptase-polymerase chain reaction（RT-PCR）. The ICAM-1 protein expression was determined by enzyme-linked immunosorbent essay（ELISA）. Western blotting method was used to detect the expression of protein kinase B（Akt）. Results 11,12-EET caused the minor injury in normal oxygen incubated HUVECs,but in hypoxia/reoxygenation HUVECs,it raised the cell viability markedly,decreased MDA content,increased the activity of SOD and the expression of Akt,and depressed the expression of ICAM-1 mRNA and protein. Conclusion 11,12-EET may provid the protective effection against endothelial cell hypoxia/reoxygenation injury. The mechanism may be related to increasing the activity of SOD and elimination of oxygen-derived free radicals,increasing the expression of Akt,and depressing the expression of ICAM-1 mRNA and protein.

关 键 词：11 12-环二十碳三烯酸 内皮细胞 缺氧/复氧损伤 

分 类 号：R541.4[医药卫生—心血管疾病]