人血管内皮生长因子165基因联合间充质干细胞治疗对扩张型心肌病胶原重塑的影响  

Collagen Remodeling Effect of Transplantation Mesenchymal Stem Cells with a Cardiac-Specific Expressing Human Vascular Endothelial Growth Factor-165 Gene in Dilated Cardiomyopathy Animal Model

作　　者：于勤[1] 朱宁[2] 李倩晓[3,4] 李晓菲[3,4] 那荣妹[3,4] 郑晓群[5] 方唯一[6] 

机构地区：[1]大连大学附属中山医院心内科,辽宁大连116001 [2]大连医科大学附属二院心内科,辽宁大连116023 [3]大连大学附属中山医院 [4]遵义医学院,辽宁大连116001 [5]大连市中心医院心内科,辽宁大连116023 [6]上海市胸科医院心内科,上海200030

出　　处：《中国临床医学》2011年第5期587-590,共4页Chinese Journal of Clinical Medicine

基　　金：辽宁省教育厅高等学校科研计划项目(编号:L2010023);大连市科技局科研计划项目(编号:2007E21SF203)

摘　　要：目的:探讨人血管内皮生长因子165基因(hVEGF165)联合间充质干细胞(mesenchymalstemcells,MSCs)治疗对扩张型心肌病胶原重塑的影响。方法:将40只扩张型心肌病(DCM)大鼠模型随机分为4组:PBS组、MSCs心肌移植联合基因治疗组(MSCs-GENE组)、MSCs心肌移植组(MSCs组)以及基因治疗组(GENE组)。心肌局部注射所构建MLC-2v/pIRES2-EG-FP-hVEGF165与MSCs,采用逆转录-聚合酶链反应(RT-PCR)检测I型、Ⅲ型胶原和转化生长因子-β(TGF-β)基因表达,蛋白免疫印迹(Westren blot)检测hVEGF165蛋白表达。结果:PBS组、MSCs+GENE组、MSCs组以及GENE组的I型胶原PCR产物灰度比分别为(0.359±0.010)、(0.240±0.012)、(0.313±0.058)、(0.230±0.011);而Ⅲ型胶原分别为(0.831±0.011)、(0.842±0.015)、(0.917±0.058)、(0.688±0.015);TGF-β1分别为(0.548±0.067)、(0.370±0.012)、(0.561±0.014)、(0.369±0.098);MSCs+GENE组TGF-β1与GENE组比较差异无统计学意义,但两者显著低于PBS组和MSCs组,提示TGF-β1表达下调;I型/III型胶原灰度比分别为(0.436±0.072)、(0.290±0.023)、(0.337±0.021)、(0.333±0.011),其中MSCs组与GENE组比较差异无统计学意义;MSCs组与GENE组的I型/III型胶原灰度比减低,2组比较差异无统计学意义,但MSCs-GENE组进一步使I型/III型胶原灰度比减低;TGF-β1表达分析结果显示,hVEGF165基因治疗较MSCs移植使TGF-β1表达下调的作用更显著。结论:MSCs移植与hVEGF165基因治疗有可能通过下调TGFβ1表达,降低I型/III胶原比值,增加心肌顺应性,减轻心肌胶原网络重塑而改善心功能。Objective：To investigate the collagen remodeling effect of transplantation mesenchymal stem cells （MSCs） combined with a cardiac-specific expressing human vascular endothelial growth factor-165 （hVEGF165） gene in dilated cardiomyopathy （DCM） animal model. Methods.. The bone marrow-derived mesenehymal stem cells of rat were isolated, cultured and expanded evivo. The characteristics of those cells were identified by flow cytomery. The subjects were divided into PBS group, MSCs transplantation group （MSCs）, gene therapeutic group （GENE）, and MSCs transplantation combined with gene transfection group （MSCs-GENE）, in which the MSCs and/or pmlc-2v-EGFP-hVEGF165 were injected into the rats＇ heart. After 1 month, reverse transcriptase PCR （RT-PCR） was used to detect the mRNA expressions of type I, type III collagen and human transforming growth factor β1 （TGF-β1）. The expressions of hVEGF165 were documented by Western blot. Resuits： Although the optical densities of TGF-β1 were significantly lower in MSCs-GENE group and GENE group than those in PBS group and MSCs, there were no difference between MSCs-GENE group and GENE group （P = 0. 910）, which suggested the expressions of TGF-β1 were down-regulation in both MSCs-GENE group and GENE group. Comparison of optical density ratio of type I/III collagen, the optical densities ratios were lower in MSCs-GENE group than that in MSCs group and GENE group, but there was no statistics different （P = 0. 172）. Though there were expressions of GAPDH in each group, but the expression of hVEGF16s were detected in MSCs-GENE group and GENE group, which suggested the target gene specifically expressing its products in myocardial. Conclusions： The mechanism of cardiac function improved by MSCs with VEGF genes may be related to the down-regulation of TGF-β1 expression, decreasing ratio of type I/III collagen, and increased myo cardial compliance and attenuation of remodeling of myocardial collagen lattice.

关 键 词：人血管内皮生长因子165基因 胶原重塑 转化生长因子Β1 间充质干细胞 基因治疗 扩张型心肌病 

分 类 号：R542.2[医药卫生—心血管疾病]