Ad-hVEGF165通过调控一氧化氮系统逆转同型半胱氨酸对内皮细胞的损伤  

Ad-hVEGF165 reverses homocysteine-induced endothelial dysfunction by regulating nitric oxide system

作　　者：赵莉莉[1] 毛用敏[1] 张莹[1] 师莹[1] 宋衍秋[1] 

机构地区：[1]天津市心血管病研究所天津市胸科医院,300222

出　　处：《中华心血管病杂志》2015年第5期432-436,共5页Chinese Journal of Cardiology

基　　金：天津市卫生局科技基金（10KG123）

摘　　要：目的 探讨携带人血管内皮生长因子165基因的重组腺病毒（Ad-hVEGF165）对经同型半胱氨酸（Hcy）损伤的内皮细胞的作用及机制.方法 不同浓度Hcy（0、0.05和1.00 mmol/L）刺激人脐静脉内皮细胞CRL-1730 24 h后,分别加入Hcy、空载体重组腺病毒Ad-Track和Ad-hVEGF165作用48 h（分为9组,包括空白组、Hcy0.05组、Hcy1.00组、Ad-Track组、Hcy0.05＋ Ad-Track组、Hcy1.00＋Ad-Track组、Ad-hVEGF165组、Hcy0.05＋ Ad-hVEGF165组、Hcy1.00＋ Ad-hVEGF165组）,采用实时荧光定量PCR和Western blot半定量分析细胞中内皮型一氧化氮合酶（eNOS）和二甲基精氨酸二甲胺水解酶（DDAH）2的mRNA和蛋白相对表达量,二者mRNA和蛋白表达的相关性采用Pearson相关分析.结果 Hcy0.05组和Hcy0.05＋Ad-hVEGF165组eNOS的mRNA和蛋白表达均分别低于空白组和Ad-hVEGF165组（P均＜0.05）.Hcy0.05组、Hcy1.00组与空白组相比,Hcy0.05＋Ad-Track组、Hcy1.00＋ Ad-Track组与Ad-Track组相比,Hcy0.05＋Ad-hVEGF165组、Hcy1.00＋Ad-hVEGF165组与Ad-hVEGF165组相比,细胞内的DDAH2 mRNA和蛋白表达均较低（P均＜0.05）.Ad-hVEGF165组与空白组、Ad-Track组相比,Hcy0.05＋Ad-hVEGF165组与Hcy0.05组、Hcy0.05＋Ad-Track组相比,Hcy1.00＋ Ad-hVEGF165组与Hcy1.00组、Hcy1.00＋ Ad-Track组相比,DDAH2 mRNA和蛋白表达均较高（P均＜0.05）.Pearson相关分析显示,Hcy损伤内皮细胞后DDAH2与eNOS的mRNA及蛋白表达均不相关（r值分别为0.057和0.449,P均＞0.05）;在外源性血管内皮生长因子作用下,DDAH2与eNOS的mRNA和蛋白表达也均不相关（r值分别为0.284和0.432,P均＞0.05）.结论 Ad-hVEGF165调控一氧化氮系统功能,是其逆转Hcy导致的内皮细胞损伤的可能作用机制之一.Objective To investigate the therapeutic effect of Ad-hVEGF165 on the endothelial cells dysfunction induced by homocysteine （Hcy） and related molecular mechanisms.Methods Human umbilical vein endothelial cells CRL-1730 were treated with Hcy at different concentrations （0,0.05,1.00 mmol/L） for 24 h.The same concentration of Hey,Ad-Track and Ad-hVEGF165 were added to the cells in the following groups：blank group,Hcy0.05 group,Hcy1.00 group,Ad-Track group,Hey0.05 ＋ Ad-Track group,Hcy1.00 ＋ Ad-Track group,Ad-hVEGF165 group,Hey0.05 ＋ Ad-hVEGF165 group,Hcy1.00 ＋ AdhVEGF165 group for 48 h.The mRNA and protein expressions of eNOS and DDAH2 were detected by realtime PCR and Western blot.The correlations of mRNA and protein expressions between endothelial nitric oxide synthase （eNOS） and dimethylarginine dimthylaminohydrolase （DDAH）2 were evaluated by Pearson correlation analysis.Results Compared with blank group and Ad-hVEGF465 group,the mRNA and protein expressions of eNOS were decreased in Hcy0.05 group and Hcy0.05 ＋ Ad-hVEGF165 group （both P ＜ 0.05）,and the mRNA and protein expressions of DDAH2 in cells treated with 0.05 mmol/L and 1.00 mmol/L Hcy were reduced as well（all P ＜ 0.05）.DDAH2 mRNA and protein expression are increased （all P ＜ 0.05） in Ad-hVEGF165 group compared with the blank group and Ad-Track,Hcy0.05 ＋ Ad-hVEGF165 and Hcy0.05 group compared with Hcy0.05 ＋ Ad-Track group,Hcy1.00 ＋ Ad-hVEGF165 and Hcy1.00 group compared with Hcy1.00 ＋ Ad-Track group.The mRNA and protein expressions of eNOS and DDAH2 were uncorrelated under the effect of Hcy（r =0.057 and 0.449,both P ＞0.05） and VEGF （r =0.284 and 0.432,both P ＞ 0.05）.Conclusion Recombinant adenovirus Ad-hVEGF165 could reverse Hcy-induced endothelial cells dysfunction via upregulating the expressions of eNOS and DDAH2.

关 键 词：血管内皮生长因子类 高半胱氨酸 一氧化氮 

分 类 号：R450[医药卫生—治疗学]