The Research On The Expression Of Calcium-activated Potassium And Chloride Channels In ICCs In The Diabetic Bladder

Background and objectiveDiabetes,which was a chronic,systemic metabolic disease,had become a global health problem.The incidence of it in China was 11.6 %,and in the United States,it also affected the 8.3% population of the US.Diabetic Bladder Dysfunction(DBD)was one of the diabetic urological complications,that affected more than 50% of diabetic patients.Early onset of occult Diabetes patients and clinicians often do not attach importance to DBD in the early stage until it came to the late stage of DBD disease.Because detrusor contraction of the patients weakened,patients suffer from Overactive Bladder(OAB),urinary retention,urinary urgency,dysuria and the declining feeling of urination,which seriously affect the quality of life.At present,the study of bladder excitability in DBD was mainly focused on the regulation of the nervous system.Most patients were not satisfied with the clinical treatment of DBD that used neurotrophic medicine.Related studies had confirmed that the excitability of bladder muscle was an important part of detrusor excitability changes,and the bladder ICC cells were the control center of the excitability of bladder muscle.What is more,study found that KCa/ClCa ion channel is the key to the excitability of ICC.In this study,bladder ICC and the KCa/ClCa ion channels of ICC cells,which were closely related to the excitatory and the contraction of the bladder,were the new point to explore the mechanisms of the pathogenesis of DBD.These results would help to provide new potential targets for clinical treatment.MethodsIn this study,SD rats,which were the research,objected by STZ,in this way,we got the model of bladder dysfunction of diabetes.First,ICCs were Separated by collagenase digestion of bladder tissue,and the purity were identified by PE Kit antibody,and then KCa/ClCa channel subtypes(BKCa,SKCa2,SKCa3,ClCa)expression was found by immunofluorescence and RT-PCR technique in normal ICCs and DBD ICCs.The last,in vitro experiment we tested KCa/ClCa channel subtypes expression by immunofluorescence and RT-PCR technique in normal bladder ICC cells cultured under glucose of different concentrations.Results1.the ICCs were harvested by collagenase digestion of bladder tissue were in 96% purity,met the needs of the follow-up study.2.BKCa,SKCa2,SKCa3 expression were significantly elevated in DBD ICCs than that in normal ICCs,and ClCa expression is decreased obviously in DBD ICCs than that in normal ICCs.3.With the increase of glucose concentrations,the expression of BKCa,SKCa2,SKCa3 were increased,the expression of ClCa was decreased.Conclusion1.We got the the ICCs by the different adherent ability between ICC-like cells and detrusor smooth muscle cells.2.Using the immunofluorescence and RT-PCR technique,we firstly recorded that BKCa,SKCa2,SKCa3 expression were significantly elevated in DBD ICCs than that in normal ICCs,and ClCa expression is decreased obviously in DBD ICCs than that in normal ICCs,indicating that the excitability of ICC cells in diabetic group decreased.3.Using the immunofluorescence and RT-PCR technique,we showed that high glucose is an important factor in the changes of BKCa,SKCa2,SKCa3,ClCa ion channels in the bladder ICCs,indicated that BKCa,SKCa2,SKCa3,ClCa may play an important role in the pathogenesis of DBD.