HGF/c-Met促进大鼠骨髓内皮祖细胞迁移能力  

HGF/c-Met increases the migration ability of marrow EPCs in rats

作　　者：秦廷江 杜明成[1] 王朝辉[1] 刘盈贝 何瑞靖 梁绍燕 安永[1] 

机构地区：[1]重庆医科大学附属儿童医院胸心外科儿童发育疾病研究教育部重点实验室儿童发育重大疾病国家国际科技合作基地儿科学重庆市重点实验室,重庆400014

出　　处：《基础医学与临床》2016年第12期1611-1617,共7页Basic and Clinical Medicine

基　　金：国家自然科学基金(81370432);重庆市教委科学技术研究项目(KJ1500238);重庆市自然科学基金(cstc2011jjA10096);国家临床重点专科建设项目(国卫办医函[2013]544)

摘　　要：目的探讨肝细胞生长因子（HGF）及其受体c-Met对大鼠骨髓内皮祖细胞（EPCs）迁移能力的影响及其机制。方法提取、培养和鉴定大鼠骨髓EPCs;重组腺病毒载体介导c-Met基因转染EPCs,用real-time PCR、Western blot和CCK8分别检测c-Met的表达和细胞增殖;分别用0、5、10、20和50 ng/m L HGF处理Ad-c-Met-EPCs,并通过Transwell迁移系统检测Ad-c-Met-EPCs的迁移能力;选取适宜浓度的HGF处理EPCs、Ad-GFP-EPCs和Ad-c-MetEPCs,并设置PBS对照组和磷脂酰肌醇3-激酶抑制剂LY294002组（同时加入HGF和10 g/L的LY294002）,通过Transwell迁移系统和Western blot分别检测各组细胞的迁移能力、Akt和P-Akt的表达。结果 1）Ad-c-Met-EPCs中c-Met基因与蛋白均为高表达（P〈0.05）。2）c-Met基因对EPCs的增殖无明显影响。3）HGF在0～50 ng/m L范围内呈浓度依赖性促进Ad-c-Met-EPCs迁移。4）HGF＋Ad-c-Met-EPCs组的迁移能力和P-Akt的表达显著高于对照组、HGF＋EPCs组、HGF＋Ad-GFP-EPCs组和HGF＋LY294002＋Ad-c-Met-EPCs组（P〈0.05）。结论 HGF/c-Met能够显著提高EPCs的迁移能力;HGF/c-Met可能通过PI3K/Akt信号通路促进EPCs的迁移。Objective To investigate the effect of hepatocyte growth factor（ HGF） and its receptor c-Met on the migration ability of rat bone marrow-derived endothelial progenitor cells（ EPCs） and its mechanism. Methods The EPCs were separated,obtained and identified. The Ad-c-Met-EPCs were obtained by recombinant adenovirus vector mediates c-Met transfered EPCs. The expression of c-Met and proliferation capacity of each group were detected by real-time PCR,Western blot,and CCK8 respectively. The migration ability of Ad-c-Met-EPCs with different concentrations of HGF was checked by Transwell system. The test cells were processed by proper concentration of HGF,while the control group and phosphatidylinositol 3-kinase inhibitor group（ add HGF and 10 g/L LY294002） were also setted. The migration ability,Akt and P-Akt of the each group cells were detected by Tran-swell system and Western blot. Results 1） The results of q PCR and Western blot showed that c-Met gene and protein in Ad-c-Met-EPCs had a high expression（ P〈 0. 05）. 2） The c-Met gene did not have significant impact on the appreciation capacity of EPCs. 3） HGF increased Ad-c-Met-EPCs migration with concentration increasing（ 0-50 ng/m L）. 4） The migration capacity of HGF ＋ Ad-c-Met-EPCs and P-Akt protein were significantly higher than other groups（ P〈 0. 05）. Conclusions HGF/c-Met can remarkably increase the migration capacity of EPCs. Besides,HGF/c-Met may accelerate the migration of EPCs through PI3 K/Akt signal path.

关 键 词：HGF C-MET 内皮祖细胞 细胞迁移 

分 类 号：R329.2[医药卫生—人体解剖和组织胚胎学] R363[医药卫生—基础医学]