The Role Of JNK Pathway Inhibitors On Protamine Sulfate Induced Bladder Inflammation Of Rats

Background and PurposeInterstitial cystitis, a kindle of chronic inflammatory disease involving the wholebladder wall, the main clinical manifestations are urinary urgency, lower abdominal painassociated with urinary. Its cause is still unknown. IC is still a lack of research aboutdetrusor lesions, even IC muscular morphology and even molecular biology research isalmost a blank. TGFβ plays an important role in a variety of fibrotic diseases, and MAPKpathway is an important signaling pathway involved in regulation of inflammation andfibrosis. This studyl explore of the morphology of human interstitial cystitis and theexpression levels of TGFβ, MAPK signaling pathway related proteins, and to explore theNAC and JNK pathway inhibitors for the treatment of rat model of bladder inflammationand possible mechanisms.MethodIn our study,11cases of interstitial cystitis bladder specimens and nine cases ofbladder cancer para-carcinoma tissue. Morphology was observed by H.E. staining includingepithelial defect, inflammatory infiltration; toluidine blue staining observe the number ofmast cells; Masson staining observe the bladder wall collagen deposition.Immunohistochemical staining observe expression and distribution of TGFβ, Smad2/3,p-Smad2/3, CTGF. Westernblot detects expression levels of TGFβ, Smad2/3, p-Smad2/3,CTGF, JNK, p-JNK, c-jun, pc-jun, ERK1/2, p-ERK1/2, P38, p-P38, IL-6, TNF-α.112female SD rats were randomly divided into normal control group, saline infusiongroup, protamine sulfate infusion group, protamine sulfate infusion+saline injection group,PPCES protamine sulfate injection+infusion group, SP600125injection+protaminesulfate infusion group, acetylcysteine injection+protamine sulfate infusion group. Eachgroup include16rats. All rats were treated three times a week, for a month. Then, eight ratsin each group were randomly selected to take Urodynamic testing under anesthesia,recording of bladder basic pressure, BP, bladder peak pressure PP, intermittentcontraction ICI, bladder capacity BC. Morphology was observed by H.E. staining,including epithelial defect, inflammatory infiltration; toluidine blue staining observe thenumber of mast cells;Westernblot detect expression levels of JNK, p-JNK, c-jun, pc-jun,IL-6, TNF-α.Results:1. HE staining, toluidine blue staining and Masson staining showed human interstitialcystitis of the bladder epithelial thinning, mast cell infiltration, collagen deposition betweendetrusor muscle bundles than normal tissue obviously.2. Immunohistochemical staining and Westernblot tests showed human interstitialcystitis bladder TGFβ, Smad2/3, p-Smad2/3, CTGF, p-JNK, IL-6, TNF-α expressionlevel was significantly higher than that in normal tissues (P <0.05). Westernblot testsshowed ERK1/2, p-ERK1/2expression decreased (P <0.05); P38, p-P38, JNK showed nodifference (P>0.05).3. In animal experiments, H.E. staining, toluidine blue staining NAC, SP600125canreduce bladder inflammation (P <0.05), reducing mast cell infiltration (P <0.05).Urodynamic test showed that NAC, SP600125can improve contraction intermittent ICI (P<0.05), bladder capacity BC (P <0.05), but the basic pressure of the bladder, BP, bladderpeak pressure PP showed no obvious improvement (P <0.05).4. In animal experiments: Westernblot showed NAC can reduce p-c-jun, IL-6, TNF-α(P <0.05). NAC can reduce the activity of JNK pathway to reduce the release ofinflammatory cytokines.Further, SP600125intervention group showed lower expressionof p-JNK, p-c-jun, IL-6, TNF-α than the control group (P <0.05). JNK pathway inhibitorscan reduce the release of inflammatory cytokines.Conclusion:In human interstitial cystitis, bladder fibrosis may be related to the activation of TGFβand MAPK pathway. In the rat model of protamine sulfate-induced bladder inflammation,NAC can ease bladder inflammation improve urodynamic parameters, which may relate tothe inhibition of activity of JNK pathway to reduce IL-6, TNF-α expression. JNK pathwayinhibitor,SP600125can significantly reduce the release of inflammatory cytokines,improve bladder inflammation and urodynamic parameters.