Differential Expressions Of NGF, PGP9.5 And 5-HT Receptor Subtypes In Autoimmune Interstitial Cystitis In Mice

Objective:To establish autoimmune IC/BPS mouse model which induced by UP3A65-84-Study on the expression of nerve growth factor and the distribution of nerve fibers in bladder at different stages, the expressions of 5-HT receptor subtypes in the T1 3-L2 spinal segments also be researched.Methods:6-8 weeks BALB/c female mice were immuned with the mixture of UP3A65-84 peptide and complete Freund’s adjuvant by subcutaneously multi-point inject. Normal BALB/c female mice as the blank control group, without UP3A65.84 polypeptide emulsifiers as the experimental control group. The characteristics of the model’s behavior, morphology and molecular biological were measured by mechanical pain response, HE staining, RTq-PCR method respectively. The expression of nerve growth factor in bladder at different stages were measured by immunohistochemistry and RTq-PCR method, The.distribution of PGP9.5 positive nerve fibers measured by immunohistochemistry and stereotactic technical analysis. The expressions of 5-HT receptor subtypes in the T13-L2 spinal segments were measured by RTq-PCR method.Results:UP3A65.84 peptide combined with complete Freund’s adjuvant can be successfully induced autoimmune IC/BPS mouse model. Behavioral performance measurement show increased pain response in model group, the ratios of bladder weight to body weight in all model groups were significantly increased (P<0.05) compared with the normal group and the CFA group. Bladder tissue morphology shows acute and chronic inflammatory cell infiltration, spotting and blood vessels to dilate, epithelial cell necrosis, etc. The expression of IL-17A and TNFmRNA in bladder were significant increase (P<0.05) compared with the normal group and the CFA group measured by RTq-PCR method. NGF protein were continuously expressed in the bladder epithelium cytoplasm. The expression of NGF protein and NGFmRNA in all model groups were significantly increased (P <0.05) compared with the normal group. PGP9.5 positive nerve fibers expressed in each layer of the bladder tissue, the positive expression in submucosal layer was significantly higher than the detrusor. The expression of PGP9.5 positive nerve fibers in submucosal layer were increased compared with the normal group,60d and 90d model group have a significantly higher number density of PGP9.5 positive nerve fibers compared with the normal group (P<0.05). The number density of PGP9.5 positive nerve fibers and NGF protein expression levels was positively correlated (r= 0.6448, p= 0.0070). The expressions of 5-HTR2A,5-HTR5A and 5-HTR7mRNA in the spinal cord of all model groups were significantly up-regulated (P<0.05) compared with the normalgroup, there is no significant differences between different model groupsConclusion:UP3A65-84 peptide combined with complete Freund’s adjuvant can be successfully induced autoimmune cystitis mice model. The expression of NGF and PGP9.5 increased in bladder of IC/BPS model and were positively correlated with pain sensitivity suggesting that NGF participate bladder pain-related neural circuits remodeling may through promote nerve fiber proliferation The expressions of 5-HTR2A,5-HTR5A and 5-HTR7mRNA in the spinal cord was significantly upregulated, suggesting that 5-HT system may be involved in spinal level regulation of pain in interstitial cystitis/bladder pain syndrome.