摘要
目的 :分析肝细胞生长因子 (HGF)能否动员骨髓内皮祖细胞 ,以及动员的内皮祖细胞能否参与创伤修复时的血管新生和内皮修复。方法 :将腺病毒HGF载体 (adenovirusvectorencodingHGFgene,Ad HGF)经尾静脉注射到Balb/c小鼠体内 ,用ELISA方法检测血浆HGF水平的变化 ;用流式细胞术检测外周血CD34+ 细胞含量变化 ;对外周血单个核细胞进行分离、培养 ,并对生长的细胞克隆进行内皮细胞表面标志Tie 2、vW因子的免疫组化检测。建立雌性小鼠CCl4肝损伤模型 ,静脉移植HGF处理后雄性小鼠外周血单个核细胞到其体内 ,4W后利用原位杂交技术检测新生肝组织中是否存在雄性细胞。结果 :注射Ad HGF能明显提高小鼠血浆的HGF水平 ,并使外周血中以CD34、Tie 2和vW因子等为标志的内皮祖细胞的数量显著增多。这些细胞参与肝损伤修复时的血管新生。结论 :HGF对骨髓内皮祖细胞具有明显的动员作用。
Aim: To assess whether hepatocyte growth factor recruits bone marrow-derived endothelial progenitor cells into blood circulation to participate in postnatal angiogenesis and endothelium repair. Methods: The adenovirus vector encoding HGF gene (Ad-HGF) were intravenous administrated into BALB/c mice, and then serum HGF was determined by enzyme-linked immunosorbent assay, the number of CD34 + cells in peripheral blood was assayed by flow cytometry, and the nucleated cells in peripheral blood were isolated, cultured and the endothelial cell colonies were characterized by staining with antibodies against tie-2, vWF. The carbon tetrachloride-induced liver damage model of female mice was established. The peripheral blood nucleated cells of Ad-HGF treated male mice were intravenous administrated into these mice, and 4 weeks later, in situ hybridization for the sry gene was used to identify the implanted cells in the damaged tissues. Results: Intravenous administration of Ad-HGF resulted in significant elevation of serum hepatocyte growth factor level and induced profoundly increase of endothelial progenitor cells in the peripheral blood, which were characterized by their ability to form endothelial cell colonies in culture and expression of CD34, tie-2, and vW factor. HGF-mobilized endothelial progenitors could incorporate into sites of neovascularization in a liver regeneration model. Conclusion: Hepatocyte growth factor could markedly recruit bone marrow-derived endothelial progenitor cells into blood circulation.
出处
《中国应用生理学杂志》
CAS
CSCD
北大核心
2005年第1期100-103,共4页
Chinese Journal of Applied Physiology
关键词
肝细胞生长因子
动员作用
内皮祖细胞
hepatocyte growth factor
mobilization
endothelial progenitor cells
