The Effect Evaluation Of Lentivirus-mediated SCL Gene Transfected DCP Bladder

Objective:To observe the distribution of green fluorescence in bladder, and to determine the amount of virus which got relative good effect of transfection; Preparing diabetic cystopathy guinea pig model, and observing the effect of SCL lentivirus transfection through perfusing bladder of guinea pig.To lay the foundation for further research on the clinical treatment of diabetic cystopathy.Methods : 1. Perfusing bladder transurethrally and intravenous injecting lentivirus carring green fluorescence, and getting tissues of bladder/liver/kidney/lungs for frozen sections after feeding seven days, observing the distribution of green fluorescence under confocal laser microscopy. 2. DCP blader model was constructed by single intraperitoneal injection of streptozotocin, screening 15 DCP guinea pigs and dividing into two groups for further experiment; Getting the tissues of bladder after perfusing 2d/14d/7d/28 d for frozen sections,observing the distribution of green fluorescence under confocal laser microscopy, and testing the expression of SCL gene.Results:1. The titer of lentivirus was 4×108, when the amount of lentivirus was 30μl, green fluorescence could be found under the mucosa, and green fluorescence distributed widely in muscle layer using 40μl lentivirus, even stronger green fluorescence was detected in muscle layer when the amount was 50μl; when 25μl lentivirus was injected intravenously, green fluorescence could be found in muscle layer, but it’s stronger in other tissues than in bladder.2. Twenty three DCP models was screened from sixty Holland guinea pigs(two months old).There was no green fluorescence in tissue,SCL level was slightly increased after perfusing bladder transurethrally 2 days; strong green fluorescencethe was found in all tissue layers and the level of SCL was significantly increased after perfusing 7 days and 14days; weak green fluorescencethe still existed and the level of SCL was still higher than normal level after 28 days.Conclusion: SCL gene can be mediated by lentivirus to transfect bladder of guinea pig successfully and expresses stablely;It,s possible to bring a new research direction and method for clinical treatment of DCP.