重组人Β防御素2在真皮多能干细胞中的表达及抗菌活性的测定  

Expression of recombinant human β-defensin 2 in dermal multipotent stem cells and its antiseptic activity

作　　者：李楠[1] 肖桃元[2] 粟永萍[1] 徐辉[1] 王军平[1] 宗兆文[1] 冉新泽[1] 董世武[1] 刘志君[1] 

机构地区：[1]第三军医大学军事预防医学院防原医学教研室,全军复合伤研究所,创伤,烧伤与复合伤国家重点实验室,重庆400038 [2]第三军医大学高原军事医学系中心实验室,重庆400038

出　　处：《第三军医大学学报》2006年第4期284-286,共3页Journal of Third Military Medical University

基　　金：国家重点基础研究发展规划资助项目("973项目")(2005CB522605);国家自然科学基金资助项目(30300185)~~

摘　　要：目的检测重组人Β防御素2(HUMANΒ-DEFENSIN2,HBD2)腺病毒表达载体在大鼠真皮多能干细胞(DERMALMULTIPOTENT STEM CELLS,DMSCS)中的表达,并观察重组HBD2的体外抗菌活性。方法将含有HBD2重组腺病毒转染DMSCS,RT-PCR、荧光免疫化学、WESTERN BLOTTING检测HBD2的表达情况,ELISA测定培养上清中HBD2的浓度,K-B纸片扩散法检测上清对大肠埃希菌、金黄色葡萄球菌和铜绿假单胞菌等标准菌株的杀灭效果。结果RT-PCR、荧光免疫化学和WESTERNBLOTTING的结果显示转染后HBD2可在DMSCS中有效地表达,上清中HBD2的浓度为743.6NG/ML,K-B纸片法显示HBD2对上述标准菌株有明显的杀灭效应。结论HBD2重组腺病毒表达载体在DMSCS可高效表达,并对大肠埃希菌等标准菌株有杀灭效应。Objective To examine the expression of human β-defensin 2 （ hBD2 ） recombinant adenovirus expression vector in rat dermal multipotent stem cells （dMSCs） and to observe the antiseptic activity of recombinant hBD2. Methods The expression of hBD2 in dMSCs was examined by RT-PR, fluorescent immunochemistry and Western blotting, and the concentration of recombinant hBD2 in supernate was measured by ELISA. The antiseptic activity of recombinant hBD2 was assessed by K-B disc agar diffusion test. Results hBD2could be effectively expressed in dMSCs, and the concentration of recombinant hBD2 in supernate was about 743.6 ng/ml. Recombinant hBD2in supernate showed antiseptic activity. Conclusion Recombinant adenovirus expression vector of hBD2 could be effectively expressed in dMSCs, and the recombinant hBD2 in supernate showed obvious antiseptic effects toward some standard bacteria lines.

关 键 词：人Β防御素2 重组 真皮多能干细胞 

分 类 号：R329.24[医药卫生—人体解剖和组织胚胎学] R977.6[医药卫生—基础医学]