基础医学与临床 ›› 2016, Vol. 36 ›› Issue (12): 1611-1617.

• 研究论文 •    下一篇

HGF/c-Met促进大鼠骨髓内皮祖细胞迁移能力

秦廷江1,杜明成2,王朝辉1,刘盈贝1,何瑞靖1,梁绍燕1,安永1   

  1. 1. 重庆医科大学附属儿童医院
    2. 重庆市渝中区中山二路136号
  • 收稿日期:2016-05-10 修回日期:2016-07-01 出版日期:2016-12-05 发布日期:2016-11-29
  • 通讯作者: 安永 E-mail:3168538416@qq.com
  • 基金资助:
    国家自然科学基金面上项目;重庆市教委科学技术研究项目;重庆市自然科学基金项目;国家临床重点专科建设项目

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

  • Received:2016-05-10 Revised:2016-07-01 Online:2016-12-05 Published:2016-11-29
  • Supported by:
    the National Natural Science Foundation of China

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的表达和细胞增值;分别用0、5、10、20和50 ng/mL 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-Met-EPCs,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 ①Ad-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的增值无明显影响。③HGF在0~50 ng/mL范围内呈浓度依赖性促进Ad-c-Met-EPCs迁移。④HGF+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。

关键词: HGF, c-Met, 内皮祖细胞, 细胞迁移

Abstract: 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 cells were detected by real-time PCR, Western blot, and CCK8 respectively. The migration ability of Ad-c-Met-EPCs with different concentrations of HGF were checked by Transwell system. The each group cells were processed by proper concentration of HGF, while the PBS control group and phosphatidylinositol 3-kinase inhibitor group (add HGF and 10g/L LY294002) were also setted. The migration ability, Akt and P-Akt of the each group cells were detected by Transwell system and Western blot. Results 1)The results of QPCR and Western blot showed that c-Met gene and protein in Ad-c-Met-EPCs had high expression(P<0.05). 2)The c-Met gene did not have significant impact on the appreciation capacity of EPCs. 3)HGF could increased Ad-c-Met-EPCs migration with concentration increasing(0-50 ng/ml). 4)The migration capacity of HGF+Ad-c-Met-EPCs and P-Akt protein were significantly higher than other groups(P<0.05). Conclusion HGF/c-Met can remarkably increase the migration capacity of EPCs. Besides, HGF/c-Met may accelerate the migration of EPCs through PI3K/Akt signal path.

Key words: Hepatocyte growth factor, c-Met, endothelial progenitor cells, cell migration

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