Therapeutic angiogenesis induced by human hepatocyte growth factor gene in rat hindlimb of ischemia / 中华老年医学杂志

ABSTRACT

Objective To investigate the effect of plasmid pEGFP- hepatocyte growth factor (HGF)-C1 on rat acute ischemia of hindlimb.Methods The eukaryotic expressed plasmid pEGFPHGF-C1 carrying human HGF cDNA was constructed.The transfection efficiency and the expression level of HGF were evaluated by transfecting pEGFP-HGF-C1 into primary rat skeletal muscle cells.Ligation-induced ischemia of femoral artery of one hindlimb in Wistar rats was performed.The plasmids (200 μg/500 μl) were injected once directly into the ischemic limb muscle (5 sites around ligation position) immediately after ligation.HGF expression was detected in the muscle tissue of rats on days 3,6 and 9 by immunochemical stain and Western blot.At day 24,the muscles were removed and stained with CD31 to assess histologically the capillary formation.Results The primary rat skeletal muscle cells could be transfected 0efficiently with pEGFP-HGF-C1 using LipofectamineTM2000 (0.8％),and secreted HGF which peak concentration was (5402.0±227.9) ng/L at 4 d.HGF expression was detected clearly in muscle tissue on days 3,6,9 in pEGFP-HGF-C1 groups.Western blot semi-quantitied analysis showed the levels of HGF expression were higher than control groups on days 3,6,9(P＜0.05).A significant increase in capillary density was found in rats transfected with human HGF(10.81±2.35) as compared with sham group (6.11±0.90) and control group (5.45±0.90) at 24 d(P＜0.01).Conclusions Intramuscular injection of naked human HGF plasmid may induce therapeutic angiogenesis in rat ischemic hindlimb models,as potential therapy for peripheral arterial disease.