四氧嘧啶诱导糖尿病小鼠的胰岛素原基因治疗  

Gene Therapy for Diabetic Mice Induced by Alloxan Using Proinsulin Gene

作　　者：刘白南[1] 唐建国[1] 纪建国[1] 顾军[1] 

机构地区：[1]北京大学生命科学学院,蛋白质工程与植物基因工程国家重点实验室,北京100871

出　　处：《中国生物化学与分子生物学报》2004年第5期681-684,共4页Chinese Journal of Biochemistry and Molecular Biology

基　　金：国家重点基础研究项目‘973计划’ (No .G2 0 0 0 0 5 69和No .G2 0 0 0 0 5 70 0 9)资助~~

摘　　要：为了解决基因治疗中的安全性问题 ,和寻求一种简便而又经济的糖尿病基因治疗模式 ,采用腹腔注射四氧嘧啶的方法 ,以昆明小鼠为实验对象 ,成功地建立了糖尿病小鼠模型 .通过电穿孔的基因转移 ,将含有胰岛素原cDNA的质粒pCMV IN转移到这些小鼠的股四头肌 .反转录PCR的结果表明 ,转基因在转染部位有转录活性 ,而放射免疫分析结果却表明转基因的表达产物分泌进了小鼠的血循环系统之中 .血糖浓度的变化说明 ,这些产物具有明显的降血糖效果 .在电基因转移前对转移部位用透明质酸酶处理 ,使电激时的使用电压降低 ,增加了转移效果 ,也更加安全 .To ensure safety in gene therapy and to find a convenient and economical gene therapy model for diabetes mellitus,the diabetic model was created in KM mice by intraperitoneal injection of alloxan. A vector pCMV-IN containing proinsulin cDNA, was delivered into diabetic mice quadriceps by electroporation with pretreatment of hyaluronidase. The result from RT-PCR indicated that the transgene was transcripted in the transfected site. The outcome of radioimmunoassay suggested that the expressed products were secreted into the circulating system. Furthermore, the products showed the function of reducing blood glucose. The injection of hyaluronidase before electroporational gene transfer lowered the electrical field to 80 V/cm, resulting in a content expression level and providing a safer mode of gene transfer.

关 键 词：糖尿病 基因治疗 电穿孔基因转移 透明质酸酶 四氧嘧啶 

分 类 号：Q78[生物学—分子生物学]