Quantitative Proteomic Analysis of Differentially Expressed Proteins and Downstream Signaling Pathways in Chronic Bladder Ischemia
Quantitative Proteomic Analysis of Differentially Expressed Proteins and Downstream Signaling Pathways in Chronic Bladder Ischemia
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DOI：
10.1016/j.juro.2015.09.079
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发表时间：
2016-02-01
影响因子：
6.6
通讯作者：
Azadzoi, Kazem M.
中科院分区：
文献类型：
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作者：
Su, Ning;Choi, Han-Pil;Azadzoi, Kazem M.
目的：越来越多的证据表明，缺血可能与衰老相关的膀胱功能障碍和下尿路症状有关。我们的目标是确定慢性缺血对膀胱蛋白质组的影响，并确定下游信号通路。材料和方法：雄性SD(R)大鼠建立双侧髂动脉粥样硬化和慢性膀胱缺血模型。8周后行膀胱测压检查。然后对缺血和对照膀胱组织进行无标记定量蛋白质组学分析。应用GO(基因本体论)和IPA(Invenuity(R)路径分析)软件对膀胱缺血改变的蛋白质进行分类。Western印迹分析证实差异表达的蛋白质。结果：慢性缺血可导致逼尿肌不稳定和顺应性降低。蛋白质组学分析显示，缺血膀胱组织中共有4,277个蛋白质，对照膀胱组织中有4,602个蛋白质。在缺血膀胱中，359和66蛋白的差异表达分别超过两倍和五倍的变化。在GO分析中，差异表达的蛋白质与蛋白质分解和退化过程中的分子信号机制有关。对缺血组织的通路和网络分析表明，改变的蛋白质参与了泛素化、Nrf2介导的氧化应激反应、细胞死亡、葡萄糖代谢和细胞骨架重构。Western印迹验证了Nedd41、MPO、CA3和FKBP5等4种代表性蛋白的变化。结论：膀胱蛋白质组学改变可能为研究盆腔动脉粥样硬化中膀胱功能障碍和下尿路症状的分子机制提供新的线索。
Purpose: Growing evidence suggests that ischemia may contribute to aging associated bladder dysfunction and lower urinary tract symptoms. Our goal was to determine the effects of chronic ischemia on bladder proteomic profiles and characterize downstream signaling pathways.Materials and Methods: Bilateral iliac artery atherosclerosis and chronic bladder ischemia were created in male Sprague Dawley (R) rats. At 8 weeks cystometrograms were obtained. Ischemic and control bladder tissues were then processed for label-free quantitative proteomic analysis. GO (Gene Ontology) and IPA (Ingenuity (R) Pathway Analysis) software were used to classify altered proteins in bladder ischemia. Western blot was done to confirm differentially expressed proteins. Tissue structure was examined by transmission electron microscopy.Results: Chronic ischemia resulted in detrusor instability and noncompliance. Proteomic analysis revealed a total of 4,277 proteins in ischemic and 4,602 in control bladder tissues. In ischemic bladders 359 and 66 proteins were differentially expressed with a greater than twofold and fivefold change, respectively. On GO analysis differentially expressed proteins were associated with molecular signaling mechanisms underlying proteolysis and degenerative processes. Pathway and network analysis of ischemic tissues suggested that altered proteins are involved in ubiquitination, Nrf2 mediated oxidative stress response, cell death, glucose metabolism and cytoskeleton remodeling. Western blot verified changes in 4 representative proteins, including Nedd4l, Mpo, Ca3 and Fkbp5. Altered proteomic profile of the bladder was associated with widespread ultrastructural damage.Conclusions: Alterations of bladder proteomic profiles in ischemia may provide new insight into molecular pathways underlying bladder dysfunction and lower urinary tract symptoms in pelvic atherosclerosis.