携带肝细胞生长因子基因的减毒沙门菌促血管形成的实验研究

发布时间：2019-02-13 02:12

【摘要】：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(P0.05)。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。
[Abstract]:Objective to investigate the effect of attenuated salmonella carrying hepatocyte growth factor (HGF) gene on angiogenesis in vitro. Methods (TPH), a attenuated Salmonella strain carrying the eukaryotic expression vector of HGF gene, was constructed and transfected into human umbilical vein endothelial cells (HUVEC) in vitro to observe the expression of the target gene and its proliferative activity. To evaluate the stimulating effect of supernatant of TPH expression on the angiogenesis of chorioallantoic membrane in chicken embryo. Results the constructed TPH strain could effectively transfect HUVEC and express active HGF protein in vitro. 6 脳 10 ~ 5 HUVEC cells could express 160 ~ 190 ng HGF protein. Compared with the control group, the expression supernatant of TPH transfected cells could significantly promote the proliferation of HUVEC. Moreover, there is a dose-effect relationship. The supernatant of TPH transfected cells also stimulated the formation of small vessels in chorioallantoic membrane of chicken embryo. The number of blood vessels (133.0 卤11.5) / cm~2 was significantly higher than that in the supernatant of HUVEC cells transfected with TP (83.3 卤5.5) / cm~2 and in the control group (62.7 卤7.1) / cm~2 (P0.05). Conclusion attenuated Salmonella carrying HGF gene can effectively transfect HUVEC, promote cell proliferation and stimulate small angiogenesis, which may play an important role in wound healing and have potential application value in the treatment of gastric ulcer.
【作者单位】：兰州军区兰州总医院检验中心/甘肃省干细胞与基因药物重点实验室;
【基金】：国家自然科学基金(81273568) 甘肃省杰出青年基金(2011GS03841) 解放军总后勤部后勤科研项目(CLZ11J09)
【分类号】：R378.22

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