新生鼠输尿管部分梗阻肾脏差异蛋白质的筛选和鉴定  

Screening and identification of differential proteins in neonatal rat kidney after partial unilateral ureteral obstruction

作　　者：赵琦[1] 杨屹[1] 牛之彬[1] 王常林[1] 侯英[1] 陈辉[1] Zhao Qi;Yang Yi;Niu Zhibin;Wang Changlin;Hou Ying;Chen Hui(Department of Pediatric Surgery,Affiliated Shengjing Hospital,China Medical University,Shenyang 110004,China)

机构地区：[1]中国医科大学附属盛京医院小儿外科,沈阳110004

出　　处：《中华小儿外科杂志》2018年第10期781-786,F0004,共7页Chinese Journal of Pediatric Surgery

基　　金：国家自然科学基金资助项目（81370772）;辽宁省自然基金项目（2013021036）

摘　　要：目的筛选新生鼠输尿管部分梗阻（partial unilateral ureteral obstruction，PUUO）后不同时间点肾脏的差异蛋白质，并检测差异蛋白PRDX1在新生鼠肾脏及先天性肾积水患儿尿液中的表达。方法应用双向电泳及质谱分析技术，检测SD新生鼠PUUO后1 d、2 d、5 d以及正常对照组肾脏组织差异蛋白质的表达。应用Western blot方法检测筛查出的差异蛋白质PRDX1在PUUO组和正常对照组新生鼠肾脏组织中的表达，应用Elisa方法检测PRDX1在20例先天性肾积水患儿及20例正常对照组患儿尿液中的表达。结果46个表达量相差2倍以上的差异蛋白质点，经质谱分析和蛋白质数据库检索成功鉴定出43个蛋白质，涉及肌动蛋白的调节、能量代谢及线粒体损伤、氧化应激及内质网应激等方面。新生鼠肾脏组织中PRDX1蛋白在PUUO1d组为0.836 1±0.017，与正常对照组相比高表达，在PUUO 2 d和5 d组分别为0.541 9±0.035和0.548 8±0.021，与正常对照组相比无明显差异。PRDX1/cr.在先天性肾积水患儿的尿液中为0.119±0.056，与对照组相比明显上调，术后1个月的尿液中PRDX1/cr.的表达为0.072±0.034，与术前相比明显下降，与正常对照组相比无明显差异。结论运用双向电泳技术结合质谱分析及生物功能信息分析筛选并鉴定出43个在PUUO新生鼠肾脏中差异表达的蛋白质，这些蛋白质可能与梗阻性肾病的发生、发展有关。其中PRDX1可能在梗阻性肾病的发生、发展过程中发挥重要的作用。ObjectiveTo screen the differential proteins in neonatal rat kidney at different timepoint with partial unilateral ureteral obstruction （PUUO） and verify the expression of PRDX1 in animal model and clinical hydronephrosis.MethodsTwo-dimensional gel electrophoresis （2-DE） and MALDI-TOF/TOF MS were employed for separating and identifying differential proteins in groups of PUUO and corresponding sham kidney tissues.Western blot and enzyme-linked immunosorbent assay （ELISA） were used for verifying the protein expression of PRDX1 in rat kidney and clinical urine samples.ResultsForty-three differential proteins whose expression levels increased or decreased 〉2-fold identified by peptide mass fingerprinting were involved in regulation of actin, energy metabolism and injuries of mitochondrion, oxidative stress and endoplasmic reticulum stress, et al.In neonatal rat kidney, higher expression of PRDX1 protein in PUUO1d, PUUO 2 d and PUUO 5 d （0.836 1±0.017, 0.541 9±0.035, 0.548 8±0.021 respectively） were detected by Western blot.In clinical urine samples, the expressions of PRDX1/cr.were up-regulated in hydronephrosis and decreased significantly within 1 month postoperatively （0.119±0.056, 0.072±0.034 respectively）.ConclusionsForty-three differentially expressed proteins associated with genesis and development of obstructive nephropathy have been successfully screened from PUUO and corresponding sham kidney tissues.And PRDX1 might be involved in the development and progression of obstructive nephropathy.

关 键 词：输尿管梗阻 蛋白质组 动物实验 

分 类 号：R726.9[医药卫生—儿科]