中国医药生物技术
中國醫藥生物技術
중국의약생물기술
Chinese Medicinal Biotechnology
2015年
6期
522-527
,共6页
哈小琴%郭馨云%葛秀洁%杨迎桂%朱晓红%张萍%杨淑娟%薛荣利%张媛媛%肖娜娜
哈小琴%郭馨雲%葛秀潔%楊迎桂%硃曉紅%張萍%楊淑娟%薛榮利%張媛媛%肖娜娜
합소금%곽형운%갈수길%양영계%주효홍%장평%양숙연%설영리%장원원%초나나
肝细胞生长因子%基因转移技术%血管生成诱导剂%转化,细菌%减毒沙门菌
肝細胞生長因子%基因轉移技術%血管生成誘導劑%轉化,細菌%減毒沙門菌
간세포생장인자%기인전이기술%혈관생성유도제%전화,세균%감독사문균
Hepatocyte growth factor%Gene transfer techniques%Angiogenesis inducing agents%Transformation,bacterial%Salmonella typhimurium
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，6×105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）/cm2明显大于转染 TP的 HUVEC细胞上清组（83.3±5.5）/cm2和对照组（62.7±7.1）/cm2（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。
目的：探討攜帶肝細胞生長因子（HGF）基因的減毒沙門菌體外促血管形成的效應。方法構建攜帶 HGF基因真覈錶達載體的減毒沙門菌菌株（TPH），體外轉染人臍靜脈內皮細胞（HUVEC）後觀察目的基因錶達及其增殖活性；評價 TPH的細胞錶達上清對鷄胚絨膜尿囊膜血管生成效應的刺激效應。結果構建的 TPH菌株在體外可有效轉染 HUVEC併錶達有活性的 HGF蛋白，6×105箇 HUVEC細胞可錶達160~190 ng的 HGF蛋白，和對照組相比，TPH轉染組細胞錶達上清可明顯促進 HUVEC增殖，而且具有劑量-效應關繫；TPH 轉染組細胞錶達上清也刺激鷄胚絨膜尿囊膜小血管生成，其血管數量（133.0±11.5）/cm2明顯大于轉染 TP的 HUVEC細胞上清組（83.3±5.5）/cm2和對照組（62.7±7.1）/cm2（P<0.05）。結論攜帶 HGF基因的減毒沙門菌可有效轉染 HUVEC併促進細胞增殖，刺激小血管新生，可能在組織創麵愈閤中起重要作用，在胃潰瘍治療中有潛在的應用價值。
목적：탐토휴대간세포생장인자（HGF）기인적감독사문균체외촉혈관형성적효응。방법구건휴대 HGF기인진핵표체재체적감독사문균균주（TPH），체외전염인제정맥내피세포（HUVEC）후관찰목적기인표체급기증식활성；평개 TPH적세포표체상청대계배융막뇨낭막혈관생성효응적자격효응。결과구건적 TPH균주재체외가유효전염 HUVEC병표체유활성적 HGF단백，6×105개 HUVEC세포가표체160~190 ng적 HGF단백，화대조조상비，TPH전염조세포표체상청가명현촉진 HUVEC증식，이차구유제량-효응관계；TPH 전염조세포표체상청야자격계배융막뇨낭막소혈관생성，기혈관수량（133.0±11.5）/cm2명현대우전염 TP적 HUVEC세포상청조（83.3±5.5）/cm2화대조조（62.7±7.1）/cm2（P<0.05）。결론휴대 HGF기인적감독사문균가유효전염 HUVEC병촉진세포증식，자격소혈관신생，가능재조직창면유합중기중요작용，재위궤양치료중유잠재적응용개치。
Objective To investigate the stimulatory effect of TPH, a recombinant attenuatedSalmonella strain carrying hepatocyte growth factor (HGF) gene eukaryotic expression vector, on angiogenesisin vitro. Methods A stable strain (TPH), recombinant attenuatedSalmonella carrying HGF gene eukaryotic expression vector, was constructed. The TPH was transfected into human umbilical vein endothelial cells (HUVEC). And the express level of HGF interest protein was assessed by ELISA. Effects of the expression product after TPH transfection on HUVEC proliferation and angiogenesis were assayed by MTT and chicken embryos chorionic allantois membrane (CAM) stimulating activity, respectively. Results The TPH could be effectively transfected into HUVEC in vitro and highly express HGF interest protein. The expression concentration of HGF protein is (160 - 190)ng/ml by 6 × 105 cells. The expression supernatant after TPH transfection could significantly stimulate the proliferation of HUVEC, with a dose-effect relation, compared with the supernatant (same volume as HGF expression supernatant) from TP-transfected and control groups. The expression of HGF supernatant after TPH transfection also could significantly promote the angiogenesis of chorioallantoic membrane (CAM). The numbers of blood vessel in expression supernatant from TP-transfected group (83.3 ± 5.5)/cm2 and PBS group (62.7 ± 7.1)/cm2 were significant fewer than that in HGF expression product from TPH-transfected group (133.0 ± 11.5)/cm2 (P< 0.05). Conclusion TPH can effectively transfect HUVEC and promote the cell proliferation, stimulate angiogenesis, may play an important role in wound healing in organization, has potential application value in gastric ulcer treatment.
