| Object: A model of diabetic cystopathy(DCP)was constructed using guinea pigs to study the effect of trans-urethral infusion of Cx43 recombinant lentiviral drugs on the expression of Connexin 43(Cx43)and m RNA on the surface of the damaged bladder forceps cells,To lay the foundation for further research on the alteration of contractile function in diabetic cystopathyMethods: Eighty healthy Dutch grade guinea pigs of similar age and body weight were selected and acclimatized for 1 week,and 15 of the 80 guinea pigs were selected for regular feeding using the randomization grouping principle and set up as the normal group;the remaining 65 guinea pigs were set up as the DCP group and were given high sugar and high fat feeding for 4 weeks,fasted for 12 h,and immediately induced with 1% streptozotocin 200 mg/kg intraperitoneal injection for diabetic guinea pigs.The successful induction of the diabetic guinea pig model was screened by random blood glucose ≥16.7 mmol/L for 4 consecutive weeks,and the remaining 15 guinea pigs were kept normally.The successfully induced animal models and normal guinea pigs were kept until 10 weeks,and the guinea pigs that did not conform to the diabetes induction model were excluded;and the urodynamic test was performed after the 10 th week to screen out the guinea pigs conforming to the DCP model.37 guinea pigs conforming to the DCP model were screened,and 36 guinea pigs were random Ly selected as experimental subjects and divided into three groups according to the random grouping principle:experimental group(n=12),control group(n=12),and blank group(n=12).The experimental group was perfused with 0.2 m L of Cx43 recombinant lentivirus via urethra,the control group was perfused with0.2 m L of empty recombinant lentivirus via urethra,and the blank group was perfused with 0.2 m L of Phosphate Buffered Saline(PBS)via urethra.Three guinea pigs in each group were executed at four time points,day 2,day 7,day 14,and day 28 after successful urethral perfusion.The expression levels of Cx43 m RNA and protein were detected by q RT-PCR assay and Western-blot assay,respectively,and the expression and distribution of Cx43 protein in the bladder tissues of the three groups were observed by immunohistochemical staining.Results:(1)Thirty-seven guinea pigs models of diabetic cystopathy were induced successfully.The forced urinary muscle pressure of the diabetic cystopathy guinea pig model was significantly lower than that of the normal group(P < 0.05),and the residual urine,maximum bladder capacity,and bladder compliance were all greater than those of the normal group,with significant differences(P < 0.05)(2)After transurethral infusion of Cx43 recombinant lentivirus,the Cx43 protein expression and m RNA expression levels were significantly higher in the experimental group compared with the control and blank groups(P < 0.05),and the recombinant lentivirus had the highest Cx43 protein and m RNA expression at day 14 after transfection,and the differences in Cx43 protein and m RNA expression levels between the control and blank groups were not statistically significant(P > 0.05).Conclusion: Transurethral perfusion of Cx43 gene can be stably expressed in bladder tissue and can upregulate Cx43 m RNA and protein expression,which may be beneficial to improve the function of damaged diabetic bladder forced muscle cells... |
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