中文摘要：

目的 探讨内皮型一氧化氮合酶（eNOS）、小凹蛋白-1（CAV1）、磷脂酰肌醇3-激酶/蛋白激酶B（PI3K/Akt）信号通路在同型半胱氨酸（Hcy）促进大鼠血管平滑肌细胞（VSMCs）迁移、增殖中的作用.方法 2016年1—6月采用组织贴块法分离SDF级SD大鼠胸主动脉中膜并传代培养VSMCs,取4-7代细胞用于实验.细胞分为4组,分别为空白对照组、100μmol/L Hcy组、200μmol/L Hcy组、500μmol/L Hcy组,使用相应浓度Hcy（0、100、200、500μmol/L）进行干预培养,采用Transwell法检测VSMCs迁移能力、MTT法检测VSMCs增殖情况〔以光密度（OD）值表示〕,硝酸还原酶法测定VSMCs释放NO情况,Western blotting检测VSMCs eNOS、CAV1、PI3K、p-Akt蛋白表达情况.结果 VSMCs穿膜细胞数与Hcy浓度呈正相关（r=0.582,P=0.020）.OD值与Hcy浓度呈正相关（r=0.620,P=0.015）.100、200、500μmol/L Hcy组释放NO低于空白对照组（P〈0.05）.NO（r=-0.479,P=0.028）,eNOS蛋白表达水平（r=-0.544,P=0.033）与Hcy浓度呈负相关.NO与eNOS蛋白表达水平呈正相关（r=0.721,P=0.009）.100、200、500μmol/L Hcy组VSMCs CAV1、PI3K、p-Akt蛋白表达水平均高于空白对照组（P〈0.05）.CAV1蛋白表达水平（r=0.505,P=0.042）,PI3K蛋白表达水平（r=0.428,P=0.030）、p-Akt蛋白表达水平（r=0.389,P=0.047）与Hcy浓度均呈正相关.结论 Hcy可能通过促进CAV1蛋白表达,抑制eNOS活性和释放,活化信号通路,从而诱导的迁移和增殖,导致动脉粥样硬化.

英文摘要：

Objective To explore the effects of endothelial nitric oxide synthase （eNOS）,caveolae - 1 （CAV1） and phosphatidylinositol3 - kinase / protein kinase B （PI3K/ Akt）signaling pathway in homocystein （Hcy）promoting rat vascular smooth muscle cells （VSMCs）migration and proliferation. Methods Between January and June 2016 VSMCs were isolated from the thoracic aorta media of the SPF grade SD rats and subcultured by substrate - attached explant method,and by which 4 - 7 generations of VSMCs were obtained. Four groups were set and given 0 （blank control group）,100,200 and 500μmol/ L Hcy group,respectively to culture the VSMCs. Transwell assay was used to detect VSMCs migration,while the conditions of VSMCs proliferation were examined by MTT assay. Nitric oxide （NO）concentration in VSMCs culture supernatant fluid was examined by nitrate reductase test,and the expressions of eNOS,CAV1,PI3K and p - Akt in VSMCs were detected by Western blotting. Results The numbers of VSMCs permeating the septum were significantly positively correlated with Hcy concentrations （r = 0. 582,P = 0. 020 ） . The MTT assay showed that positive correlations were found between OD values and Hcy concentrations （r = 0. 620,P = 0. 015）. NO concentrations of 100,200 and 500 μmol/ L Hcy groups were all lower than those of the blank control group （P 〈 0. 05）. Hcy concentrations were negatively associated with NO concentrations （r = - 0. 479,P= 0. 028）as well as eNOS expressions （r = - 0. 544,P = 0. 033）. NO concentrations were positively correlated with eNOS expressions （r = 0. 721,P = 0. 009）. Expressions of CAV1,PI3K and p - Akt in VSMCs cultured in 100,200 as well as 500μmol/ L Hcy groups were all higher than those in the blank control group （P 〈 0. 05）. Hcy concentrations showed positive correlation with CAV1 expressions （r = 0. 505,P = 0. 042）,PI3K expressions （r = 0. 428,P = 0. 030 ）,and p - Akt expressions （r = 0. 389,P = 0. 047）. Conclusion Hcy may induce VSMCs migration