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目的构建一种携带人肝细胞生长因子(HGF)基因的表达质粒(pUDKH),探讨重组质粒对体外培养的成人外周血中内皮祖细胞(endothelial progenitor cell,EPC)增殖的影响。方法用PCR方法从人胎盘cDNA文库扩增HGF基因,并将其克隆至病毒表达载体pLenti6/V5-Dest,获得表达质粒pLenti-HGF。制备病毒,感染原代培养的内皮祖细胞,免疫组化分析其感染效率及上清中HGF的表达水平。MTT法检测感染病毒后对细胞增殖的影响。结果 pLenti-HGF病毒可有效感染体外培养的内皮祖细胞,表达HGF,并能促进内皮祖细胞的增殖。结论内皮祖细胞经病毒感染后可以表达hHGF,外源性hHGF基因能够刺激内皮祖细胞增殖。
Objective To construct an expression plasmid (pUDKH) carrying human hepatocyte growth factor (HGF) gene and investigate the effect of recombinant plasmid on the proliferation of endothelial progenitor cells (EPCs) cultured in vitro. Methods The HGF gene was amplified from human placenta cDNA library by PCR and cloned into the viral expression vector pLenti6 / V5-Dest to obtain the expression plasmid pLenti-HGF. The virus was infected and infected with primary cultured endothelial progenitor cells. The infection efficiency and the expression of HGF in the supernatant were analyzed by immunohistochemistry. Effect of Infected Virus on Cell Proliferation Detected by MTT Method. Results The pLenti-HGF virus can effectively infect endothelial progenitor cells cultured in vitro, express HGF and promote the proliferation of endothelial progenitor cells. Conclusions Endothelial progenitor cells can express hHGF after virus infection. Exogenous hHGF gene can stimulate the proliferation of endothelial progenitor cells.