PROTEOMIC AND BIOINFORMATIC ANALYSISOF UROTHELIUM IN OVERACTIVE RAT MODELS
Y. NA, J. SHIN, J. LIM, K. SONG, J.PARK, S. YANG;
Chungnam Natl. Univ Hosp, Daejeon, Korea,Republic of.
Introduction: The bladderurothelium has important pathophysiological role to manifestoveractive bladder (OAB) symptoms. However, the mechanisms ofpathogenesis of OAB at the molecular level remain poorly understood,mainly as a result of lack of modern molecular analysis.
Objective:In this study we tried to identify novel urothelium proteins that arerelated to the development of detrusor overactivity (DO) usingproteomic and bioinformatic methodologies.
Methods: Thestudy was conducted using male Sprague-Dawley rats, subdivided intosham operated (n=10) and partial BOO groups (n=30). Partial BOO wasinduced for 2 weeks and DO was confirmed with measuring cystometry.The urothelium was separated from muscle layer under a dissectingmicroscope and the identification of protein was assessed withLC-MS/MS using LTQ-Velos mass spectrometry. Functional analysis ofthe data set was done using the Ingenuity GO (Gene Ontology) andPathway Knowledge Base.
Results: A total of 507 proteinswere detected in normal rat urothelium and 380 proteins in OAB raturothelium. 201 and 74 proteins were expressed uniquely in normal andOAB urothelium. The results of bioinformatic analysis using IPAidentified 17 putative upstream regulators. These regulators areinvolved primarily in inflammation and cytoskeletal organization.In the OAB urothelium, pathways involved in inflammation, such asthe complement system, acute phase response signaling, LXR/RXRactivation, and p38 MAPK signaling, were notably up-regulated. Bycontrast, signaling pathways related to cytoskeletal organization,including ILK signaling, RhoA signaling, and remodeling of epithelialadherens junctions, were commonly down-regulated in OAB urothelium.It is also notable that proteins involved in the unfolded proteinresponse (ER stress) were down-regulated and proteins involved indeath receptor signaling were up-regulated. 52 proteins involved insignaling in bladder urothelium were identified; 34 may regulate theproduction or release of ATP, 18 may regulate the production andrelease of NO.
Conclusions: Signaling pathway analysisrevealed that the differentially expressed proteins were mainlyinvolved in the inflammatory response and apoptosis. Ourfindings will provide new insights understanding the development andpathophysiology of OAB.
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