Effect Of Transurethral Bladder Instillation Of SCL And Cx43 Gene Lentivirus On DCP Function And Exploration Of Mechanism

Object: To investigate the effects and mechanisms of the combination of Stem Cell Leukemia(SCL)gene and Gap Junction Protein-43(Connexin43,Cx43)gene on the function of diabetic cystopathy(DCP)in guinea pigs.Methods: Ninety healthy male guinea pigs aged 2～3 months were fed with diets and vegetables for 1 week,and then given a single intraperitoneal injection of streptozotocin(STZ)at 200 mg/kg,and random blood glucose was monitored regularly every week.After 12 weeks of normal feeding,40 guinea pigs meeting the criteria for DCP were screened by urodynamic examination and randomly divided into 4 groups:(1)diabetic group,(2)SCL lentivirus group,(3)Cx43 lentivirus group,and(4)SCL+Cx43 lentivirus group.The diabetic group instilled 0.2 ml of empty lentivirus without gene into the bladder via urethra,the SCL group and Cx43 groups instilled 0.2 ml of SCL lentivirus and Cx43 lentivirus respectively using the same method,and the SCL+Cx43 lentivirus group instilled 0.2 ml each of SCL and Cx43 lentivirus via urethra.Urodynamic examinations were performed 14 days after transfection to assess bladder function.After the examination,the guinea pigs were executed and the bladders were quickly removed and bladder muscle strips were made to assess the contraction frequency and amplitude of the muscle strips in isolation.Finally,frozen sections of bladder tissue were prepared for fluorescence double-staining using laser confocal microscopy to observe changes in Interstitial Cells of Cajal(ICC)and Cx43.Results: 1,Transurethral instillation of SCL gene lentivirus and Cx43 gene lentivirus could be successfully transfected into guinea pig bladder with stable expression.2,There was no difference in urodynamic examination results between SCL and Cx43 groups compared with diabetic group(P < 0.05),and urodynamic examination of SCL+Cx43 group compared with diabetic group showed improvement in forced urinary muscle pressure,abdominal pressure and bladder pressure(P > 0.05).3,In the isolated forceps muscle strip experiment,the amplitude and frequency of forceps contraction were significantly decreased in the diabetic group compared with the normal group(P < 0.05),the amplitude and frequency of forceps contraction were improved in the SCL group compared with the diabetic group(P < 0.05),and there was no difference in the amplitude and frequency of forceps contraction in the Cx43 group compared with the diabetic group(P >0.05)There was no difference between the SCL and Cx43 groups(P > 0.05).4,The number of ICC was reduced in the diabetic group by laser confocal,and the cellular shuttle structure and cellular In the SCL and Cx43 groups,the number of ICC-like cells was increased,the cell shuttle structure and cell protrusion were improved,and ICC-dimers could be formed.Conclusion: Combined transurethral instillation of SCL gene lentivirus and Cx43 gene lentivirus improved the contraction amplitude and frequency of isolated detrusor muscle,and the improvement was more significant than that of single instillation.The improvement of urodynamic examination indexes such as forced urinary muscle pressure,abdominal pressure,and bladder pressure was also better with combined instillation than with single instillation.The increased number of ICC cells and improved cell structure were observed by laser confocal microscopy,and it was observed that similar ICCs could form ICC-dimers.We speculate that the formed ICC-dimer structure is an important basis for the effect of combined perfusion,and the specific mechanism needs to be further investigated.