The Mechanisms Of Endoplasmic Reticulum Stress Inhibiting Autophagy Flux To Increase The Excitability Of Bladder In Rat Interstitial Cystitis Model

Background and PurposeInterstitial cystitis is a pathogenesis unknown Clinical yndrome,The major characte ristic is urgency,frequency,suprapubic or pelvic pain,post-voiding bladder filling could reduce the performance of clinical syndrome.It is also known as painful bladder syndrome or bladder irritation syndrome,its pathogenesis is unknown to date.Autophagy plays an important role in the pathogenesis of interstitial cystitis,studies have confirmed that ER stress participated in interventing autophagy flux in a variety of inflammatory diseases and intervened occurrence,development of these diseases,this study should be adopted by the endoplasmic reticulum stress regulating autophagy flux angle to clarify the pathogenesis of interstitial cystitis.Method1.In this study,we used SD rats as the research object,first we created two groups control(C),interstitial cystitis(IC)rat model,we respectively applicated Western blot to observe the autophagy flux markers(P62),autophagy relative markers(LC3A / B,Beclin1),endoplasmic reticulum stress markers(GRP78).To clear ER stress,autophagy level and to confirm autophage flux was blocked or not in interstitial cystitis rat model.2.The first step was to clear the high level of endoplasmic reticulum stress and autophagy in interstitial cystitis,and autophagic flux was blocked when we used endoplasmic reticulum stress inhibitor 4-PBA,to further establish control(C),interstitial cystitis(IC),interstitial cystitis + 4-PBA(IC + 4-PBA)group.3.Molecular biology: The expression of endoplasmic reticulum stress marker(GRP78),autophagy flux markers(P62)and autophagy markers(LC3A / B,Beclin1)were detected by Western blot in three groups.4.Histological examination: We used immunofluorescence to detect the expression levels of LC3 A / B rats bladder tissue of these three groups.Toluidine blue staining was used to detect the mast cell invasion situation in bladder tissue.Then HE staining was detected the epithelial tissue integrity,inflammatory cell infiltration and tissue edema in bladder tissue of these three groups.5.Bladder function detecting: were We observed intermittent contractions and bladder voiding frequency changes by urodynamic testing technology in order to assess bladder function of these three groups rats.Results:1.Western blot analysis results of C and IC Group showed that the ER stress and autophagy flux marker GRP78,P62 in IC group was significantly higher than in C group,autophagy markers LC3 A / B,Beclin1 were also significantly higher in IC group than in C group(P <0.05).2.We used endoplasmic reticulum stress inhibitor 4-PBA to intervent IC group,the results Western blot analysis of GRP78,P62,LC3 A / B,Beclin1 in C,IC and IC + 4-PBA groups were found that the expression of GRP78,P62,LC3 A / B,Beclin1 in IC + 4-PBA group was significantly lower than in IC group and higher than in group C(P <0.05)3.Immunofluorescence tested LC3 A / B protein expression in bladder tissues and we found the expression was significantly lower in IC + 4-PBA group than in IC group and higher in IC + 4-PBA group than in group C(P <0.05)4.HE staining showed that the bladder tissue was significant epithelial injury,inflammatory cell infiltration,mucosa and lamina propria edema in IC group,and in IC + 4-PBA group showed better integrity of epithelial tissue,tissue edema and inflammatory cell infiltration was significantly reduced(P <0.05)5.Toluidine blue staining results showed that mast cell infiltration was significantly higher in IC group than in the control group,and in IC + 4-PBA group showed fewer bladder tissue mast cell invasion.6.Urodynamic results showed the IC group rats exhibited intermittent bladder contractions shortened and frequency of urination increased compared with C group.The bladder function was improved in IC + 4-PBA group rats: intermittent contractions increased,urination frequency was significantly lower(P <0.05).Conclusion:1.High level of the in interstitial cystitis bladder tissue and autophagy flux was blocked.2.We used endoplasmic reticulum stress 4-PBA to intervene IC group,amd we found that the inhibition of endoplasmic reticulum stress,autophagy flux being restored and the autophagy level was reduced.3.Immunofluorescence results further confirmed that inhibition of endoplasmic reticulum stress could reduce autophagy level.4.HE result indicated that the bladder tissue was improved by inhibiting endoplasmic reticulum stress to recover autophagic flux.5.Toluidine blue staining results showed that inhibiting endoplasmic reticulum stress could inhibit bladder inflammation.6.Urodynamic results confirm that inhibition of endoplasmic reticulum stress recovering autophagy flux could restore the bladder function.