经尿道灌注慢病毒转染豚鼠膀胱的研究  

The study of transfection through perfusing bladder of guinea pig with lentivirus

作　　者：魏艳青[1] 王江平[1] 王勤章[1] 钱彪[1] 丁国富[1] 

机构地区：[1]石河子大学第一附属医院泌尿外科,832000

出　　处：《天津医药》2015年第11期1275-1277,共3页Tianjin Medical Journal

基　　金：国家自然科学基金资助项目(81360120)

摘　　要：目的观察慢病毒介导绿色荧光蛋白(GFP)转染膀胱的效果,并确定取得较好转染效果的病毒量。方法豚鼠经尿道灌注及静脉注射不同量的携带GFP基因的慢病毒,饲养7 d后取膀胱、肝、肺、肾等组织冰冻切片,激光共聚焦显微镜下观察各组织GFP分布。结果滴度为4×108的慢病毒,经尿道膀胱灌注30μL时可见组织黏膜下有绿色荧光,40μL时组织肌层有广泛绿色荧光分布,50μL时组织肌层有更强绿色荧光分布,静脉注射25μL时组织肌层可见有广泛绿色荧光分布;经尿道灌注各病毒量下未在膀胱外组织见有明显绿色荧光分布,静脉注射下在肝、肺等组织见有较膀胱更强绿色荧光。结论慢病毒可以成功介导GFP转染豚鼠膀胱,并且经尿道灌注可以避免静脉注射带来的病毒在膀胱外组织广泛分布,能够为膀胱疾病的临床治疗带来新的研究方向和手段。Objective To observe the effects of green fluorescence protein mediated by lentivirus in bladder, and to de？termine the amount of virus obtained good transfection effects. Methods lentivirus carring GFP gene was perfused using transurethral approach into bladder of guinea pigs. Samples of bladder, liver, kidney and lungs were collected for frozen sec-tions after feeding seven days. The distribution of green fluorescence was observed using laser confocal microscopy. Re？sults The titer of lentivirus was 4 × 108. GFP was found under the mucosa when the amount of lentivirus transurethral was 30μL. GFP was distributed widely in muscle layer using 40μL lentivirus. GFP was detected even stronger in muscle layer when the amount was 50μL. GFP was found in muscle layer when 25μL lentivirus was injected intravenously. GFP was not found in other tissues than in bladder via transurethral perfusion. There was higher GFP in liver, lungs and other organs than in bladder via intravenous injection. Conclusion Lentivirus can mediate GFP transfecting bladder of guinea pig successful？ly and escape the distribution of GFP all over the body intravenously, which will bring new research direction and method for clinical treatment of diseases in bladder.

关 键 词：慢病毒感染 膀胱 转染 绿色荧光蛋白质类 豚鼠 

分 类 号：R694[医药卫生—泌尿科学]