经尿道灌注慢病毒转染豚鼠膀胱的研究

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 30 μL 时可见组织黏膜下有绿色荧光， 40 μL 时组织肌层有广泛绿色荧光分布， 50 μL 时组织肌层有更强绿色荧光分布， 静脉注射 25 μL 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。

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

Objective To observe the effects of green fluorescence protein mediated by lentivirus in bladder, and to determine 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 sections after feeding seven days. The distribution of green fluorescence was observed using laser confocal microscopy. Results The titer of lentivirus was 4×108. GFP was found under the mucosa when the amount of lentivirus transurethral was 30 μL. GFP was distributed widely in muscle layer using 40 μL lentivirus. GFP was detected even stronger in muscle layer when the amount was 50 μL. GFP was found in muscle layer when 25 μL 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 successfully 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.