| Background:Erectile dysfunction is a common complication of diabetes.High levels of oxidative stress are characteristic of the penile cavernosa in diabetic rats.Injection of MSC into the penis sponge is a novel therapy to erectile function of diabetic rats,but it also exposed obviously inadequate that MSCs showed low activity and short survival time under high oxidative stress.It’s urgent to find a novel method to prolong the survival efficiency of MSCs and improve its therapeutic effect.Objective(1)Establishment of erectile dysfunction model in type 1 diabetic rats;(2)Study the expression of Nrf2and HO-1 antioxidant pathway in penile cavernous tissue during probucol-MSCs combination therapy for DMED;(3)Explore the expression of antioxidant pathway in oxygen;Survival status of MSCs and expression of p62,Beclinl,Nrf2 and HO-1 in cells under chemical stress or probucol intervention.(4)To investigate the effect of probucol oxidative stress on autophagy of MSCs and its mechanism.Methods:In animal experiment,52 only 12 weeks of male Sprague wrote-Dawley rats fasting 12 hours.40 rats by intraperitoneal injection of one-time STZ 60 mg/kg.Only give 1 x PBS,12 rats were as control group.After that,the random blood glucose concentrations of 36 stz-treated rats were all greater than 16.7mmol/L.According to these test results,the establishment of diabetic rats was successful.These sham group rats and diabetic rats were divided into following 4 groups randomly which are:1.Feedings of probucol at doses of 500 mg/kg/day for 4days before intracavernosal(IC)injection of MSCs(1 × 106)and treated daily with intragastric probucol at the same doses after intracavernosal injection of MSCs(P+M group,n=12);2.intracavernosal injection of MSCs(1× 106)in diabetic rats and feeding the same dose of saline(M group,n=12);3.diabetic group(DM group,n=12)and sham group feeding the same dose of saline(S group)for 3 days,1 week,and 2 weeks.After 4 weeks,the cavernous nerve was electrical stimulated to evaluate erectile function.Penile tissue was harvested and histologic/molecular analyses were performed after the number of MSCs was observed by frozen sections to evaluate the survival status after euthanasia.In cell experiments,MSCs was collected and cultured from S-D male rats.Mesenchymal stem cells were identified by flow cytometry and then expanded and cultured.The third generation cells were used for further experiments.Mesenchymal stem cells were randomly divided into the following groups:(1)PBS was added into MSCs medium as sham group;(2)H2O2 was added into MSCs medium as oxidant group;(3)Probucol was added into MSCs medium as treatment group under the intervention of H2O2.The survival status of MSCs in different oxidative stress and probucol intervention environments were detected.Results:In animal experiments,after 2 weeks of treatment,the number of viable cells in P+M group was significantly increased,and the erectile function was significantly better than that in DM group(P<0.05),Nrf2 and HO-1 expression were higher than in DM group(P<0.05).We proved that probucol combined with MSCs could significantly improve the therapeutic effect of MSCs.In cell experiments,the oxidative stress induced by H2O2 levels can obviously decrease the activity of MSCs,concentration dependence,under the cloth to take an examination of intervention on the street,with increasing concentration,cell autophagy level called oxidative stress environment raised(P<0.05),we prove that ordinary cloth can enhance cell activity,improve their survival condition in oxidative stress condition.Conclusion:① the results showed that the level of oxidative stress in penile cavernous tissue of diabetic rats with erectile dysfunction was higher than that of blank control group.The erectile function of diabetic rats treated with probucol was improved,and the level of oxidative stress was significantly lower than that of diabetic rats,and the expression of antioxidant stress protein was higher than that of diabetic rats.Rats increased significantly.Probucol improves oxidative stress in the penile cavernous tissue of diabetic rats and indirectly improves the survival of MSCs.②In oxidative stress environment,probucol can enhance the ability of antioxidant stress in MSCs.③The survival of MSCs decreased under oxidative stress,and the cell activity increased after probucol intervention,which proved that probucol could improve the survival status of MSCs under oxidative stress. |
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