肌腱愈合及粘连形成机制的研究:胰岛素样生长因子1基因真核表达载体的构建及表达  

Research on the mechanisms of tendon healing and desmoplasia:construction of eukaryotic expression vector of insulin like growth factor 1 gene and its expression in rat tendon cells

作　　者：程昌志[1] 张正治[1] 潘险峰[1] 苟俊[1] 孙玮[1] 潘峰[1] 傅晓岚[1] 白贵友[1] 

机构地区：[1]解放军第三军医大学中心实验室,重庆市400038

出　　处：《中国临床康复》2004年第20期3984-3986,i002,共4页Chinese Journal of Clinical Rehabilitation

基　　金：国家自然科学基金资助项目(39970387)~~

摘　　要：目的:探讨真核表达载体介导外源性胰岛素样生长因子-1(IGF-1)基因转染肌腱细胞的可能性。方法:组织块法培养原代肌腱细胞,体外重组真核表达载体pcDNA3.1(+)-IGF-1,并以脂多胺DOGS为介导转染培养的肌腱细胞,反转录多聚酶链反应(RT-PCR)检测IGF-1mRNA的表达,ELISA法检测IGF-1活性蛋白表达。结果:成功构建真核表达载体pcDNA3.1(+)-IGF-1;RT-PCR检测显示转染后的肌腱细胞成功表达IGF-1mRNA;ELISA结果显示转染组IGF-1蛋白表达较对照组明显增高(t=-2.873,P<0.05)。结论:重组的真核表达载体pcDNA3.1(+)-IGF-1能够将IGF-1cDNA成功导入肌腱细胞并表达。AIM: To explore the possibility of transfecting exogenous insulin like growth factor 1 (IGF 1) mediated by eukaryotic expression vector into tendon cells. METHODS:Primary tendon cells were cultured with tissue block method. Eukaryon expression vector of pcDNA3.1(+) IGF 1 was recombined in vitro and then transfered into tendon cells mediated by lipopolyamine DOGS.The expression of IGF 1mRNA was identified by RT PCR and the active protein was determined by ELISA method. RESULTS:pcDNA3.1(+) IGF 1 of eukaryon expression vector was constructed successfully.IGF 1 mRNA was successfully expressed in tendon cells after transfection. The results of ELISA showed that the content of IGF 1 active protein in the cultured medium in the experimental group was significant higher than that in control group(t=-2.873,P< 0.05). CONCLUSION:The recombinated eukaryon expression vector of pcDNA3.1(+) IGF 1 can successfully transfer IGF 1 cDNA into tendon cells and express normally.

关 键 词：胰岛素样生长因子Ⅰ 腱/细胞学 基因表达 遗传载体 转染 

分 类 号：R349.83[医药卫生—基础医学]