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目的探讨携带肝细胞生长因子(HGF)基因的减毒沙门菌体外促血管形成的效应。方法构建携带HGF基因真核表达载体的减毒沙门菌菌株(TPH),体外转染人脐静脉内皮细胞(HUVEC)后观察目的基因表达及其增殖活性;评价TPH的细胞表达上清对鸡胚绒膜尿囊膜血管生成效应的刺激效应。结果构建的TPH菌株在体外可有效转染HUVEC并表达有活性的HGF蛋白,6×10~5个HUVEC细胞可表达160~190 ng的HGF蛋白,和对照组相比,TPH转染组细胞表达上清可明显促进HUVEC增殖,而且具有剂量-效应关系;TPH转染组细胞表达上清也刺激鸡胚绒膜尿囊膜小血管生成,其血管数量(133.0±11.5)/cm~2明显大于转染TP的HUVEC细胞上清组(83.3±5.5)/cm~2和对照组(62.7±7.1)/cm~2(P<0.05)。结论携带HGF基因的减毒沙门菌可有效转染HUVEC并促进细胞增殖,刺激小血管新生,可能在组织创面愈合中起重要作用,在胃溃疡治疗中有潜在的应用价值。
Objective To investigate the effect of attenuated Salmonella typhimurium carrying hepatocyte growth factor (HGF) gene on angiogenesis in vitro. Methods The attenuated Salmonella typhimurium strain (TPH) harboring HGF gene eukaryotic expression vector was constructed and transfected into human umbilical vein endothelial cells (HUVEC) in vitro to observe the target gene expression and proliferation activity. To evaluate the effect of TPH supernatant on chick embryo Stimulation effect of chorioallantoic membrane angiogenesis effect. Results The constructed TPH strain could effectively transfect HUVECs in vitro and express HGF protein. 6 × 10 ~ 5 HUVEC cells expressed 160 ~ 190 ng HGF protein. Compared with the control group, the TPH transfection group expressed The supernatant could significantly promote the proliferation of HUVEC with a dose-effect relationship. The supernatant of TPH transfection group also stimulated the formation of chorioallantoic membrane angiogenesis in chick embryo chick embryo. The blood vessel number (133.0 ± 11.5) / cm ~ 2 was significantly greater than The HUVEC transfected TP group (83.3 ± 5.5) / cm ~ 2 and the control group (62.7 ± 7.1) / cm ~ 2 (P <0.05). Conclusion The attenuated Salmonella carrying HGF gene can effectively transfect HUVECs and promote cell proliferation and stimulate angiogenesis, which may play an important role in the healing of tissue wounds. So it has potential value in the treatment of gastric ulcer.