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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。
OBJECTIVE: To investigate whether hepatocyte growth factor (HGF) can mobilize bone marrow endothelial progenitor cells and whether mobilized endothelial progenitor cells can participate in angiogenesis and endothelial repair during wound healing. Methods: The HGF gene was injected into the Balb / c mice via the caudal vein and the level of plasma HGF was measured by ELISA. The changes of peripheral blood CD34 + cells were detected by flow cytometry. Blood mononuclear cells were isolated, cultured, and immunohistochemical detection of endothelial cell surface markers Tie 2 and vW was performed on the growing cell clones. The CCl4 hepatic injury model was established in female mice. The peripheral blood mononuclear cells of male mice were transplanted into the body of HGF mice after intravenous administration. After 4 weeks, the presence of male cells in the newborn liver tissue was detected by in situ hybridization. Results: Ad HGF injection significantly increased HGF level in plasma and markedly increased the number of endothelial progenitor cells marked by CD34, Tie 2 and vW in peripheral blood. These cells are involved in angiogenesis when liver damage is repaired. Conclusion: HGF has a significant mobilization effect on endothelial progenitor cells of bone marrow.