过表达c-Met蛋白诱导内皮祖细胞分布于肺动脉高压大鼠肺组织  

Overexpression of c-Met protien mediates EPCs distribution in lung of hypoxia pulmonary arterial hypertension rat

作　　者：杜明成[1] 秦廷江 夏世辉 王朝辉[1] 刘赢贝 安永[1] 

机构地区：[1]重庆医科大学附属儿童医院儿童发育疾病研究教育部重点实验室认知发育与学习记忆障碍转化医学重庆市重点实验室,重庆400014

出　　处：《基础医学与临床》2016年第5期644-650,共7页Basic and Clinical Medicine

基　　金：国家自然科学基金(81370432);重庆市自然科学基金(No cstc2011jj A10096);国家临床重点专科建设资助项目(国卫办医函[2013]544)

摘　　要：目的探讨过表达c-Met蛋白对内皮祖细胞(EPCs)在低氧性肺动脉高压(HPAH)大鼠中的分布情况。方法复制HPAH模型,原代培养大鼠骨髓源性EPCs并鉴定,构建带有c-Met基因的腺病毒并转染EPCs,细胞分为对照组、空载病毒组和c-Met过表达组,用real time PCR和Western blot检测c-Met mRNA和蛋白表达,用CM-Dil活细胞染色剂标记EPCs,通过尾静脉将各处理组EPCs输注到HPAH大鼠模型体内,1周后观察EPCs在主要脏器中的分布。结果对照组、空载病毒组和c-Met过表达组c-Met mRNA表达量分别为:0.19、0.17和1.85;c-Met蛋白的表达量分别为:0.24、0.17和0.82。正常EPCs组中细胞在肺脏、肝脏、脾脏和肾脏的阳性率分别是:1.7%、0.5%、7.9%和9.4%,空载病毒组细胞阳性率分别是:1.5%、0.4%、8.3%和8.5%,c-Met EPCs组细胞阳性率分别为9.3%、0.2%、1.3%和0.3%。c-Met EPCs组中肺脏的细胞阳性率明显升高,在肾脏和脾脏的分布明显下降(P<0.001)。结论过表达c-Met蛋白促进EPCs优势分布于HPAH模型的肺组织。Objective To discuss overexpression c-Met protein mediated distribution of EPCs in HAPH model. Methods To establish the model of HPAH, primary culture rat bone marrow-derived EPCs and identificated, adenoviral with c-Met gene infected EPCs and Cells were divided into control group, viral load group, c-Met over-expression group, c-Met expression in protein and gene level were detected by Western blot and real time PCR, endothelial progenitor cells were marked by CM-Dil, the cells were infused through the tail vein of rat, distribution of the cells were observed in of the major organs afte 1 week. Results c-Met mRNA expressions of control group, viral load group and c-Met over-expression group were 0. 19,0. 17 and 1.85 ;c-Met protein expressions were 0. 24,0. 17 and 0. 82. The positive rates of EPCs in lungs,liver, spleen and kideny of EPCs group were 1.7% ,0.5% ,7.9% and 9.4% ;The positive rates of EPCs in viral load group were：l. 5% ,0. 4% ,8.3% and 8.5% ;The positive rates of EPCs in c-Met EPCs group were 9.3% ,0. 2% , 1.3% and 0. 3% ,The positive rate of EPCs in c-Met EPCs group significant higher in lung and decreased in spleen and kideny compared with control group （ P 〈 0. 001 ）. Conclusions Overexpression of c-Met protein makes EPCs superiority distributed in lung of HPAH model.

关 键 词：低氧性肺动脉高压 c—Met蛋白 内皮祖细胞 细胞分布 

分 类 号：R544.1[医药卫生—心血管疾病]