HGF基因对大鼠血管内皮细胞增殖的作用

Effect of HGF gene on proliferation of vascular endothelial cells in rats

目的构建肝细胞生长因子(HGF)真核细胞表达质粒pEGFP-HGF-C1,探讨HGF对大鼠血管内皮细胞(VECs)的作用。方法通过亚克隆技术,构建含有人HGF cDNA全长的真核细胞表达质粒pEGFP-HGF-C1并测序;向大鼠原代骨骼肌细胞转染质粒pEGFP-HGF-C1,测定转染效率;ELISA法测定细胞上清液中HGF蛋白表达浓度;MTT法测定HGF表达产物对大鼠VECs促增殖作用。结果成功构建正向表达HGF的真核细胞表达质粒pEGFP-HGF-C1,该质粒可有效转染原代培养的骨骼肌细胞并表达HGF,且其表达产物HGF对大鼠VECs的促增殖作用具有量效和时效关系(P<0.05或P<0.01)。结论成功构建的真核表达质粒pEGFP-HGF-C1能有效转染大鼠骨骼肌细胞,分泌的HGF对大鼠VECs有促增殖作用。

Objective To construct the eukaryotic expression plasmid pEGFP-HGF-C1 of hepatocyte growth factor（HGF） and investigate the effect of HGF on vascular endothelial cells（VECs） in rats. Methods The eukaryotic expression plasmid pEGFP-HGF-C1 with whole length of human HGF cDNA was constructed by subcloning technique and then sequenced.The transfection efficiency was evaluated by transfecting pEGFP-HGF-C1 into primary skeletal muscle cells in rats,and the protein expression of HGF in the cell supernatant was detected by ELISA.The proliferative effect of HGF-expressing product on VECs was measured by MTT assay. Results The pEGFP-HGF-C1 expressing HGF was successfully contructed and effectively transfected into primary skeletal muscle cells to express HGF. The HGF-expressing product could significantly stimulate the proliferation of VECs in dose-and time-dependent manner（P0.05 or P0.01）. Conclusion The eukaryotic expression plasmid pEGFP-HGF-C1 constructed can be effectively transfected into skeletal muscle cells in rats and promote the proliferation of VECs.