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大鼠单侧输尿管梗阻模型中尿液候选生物标志物的发现

Urinary candidate biomarker discovery in a rat unilateral ureteral obstruction model.

作者信息

Yuan Yuan, Zhang Fanshuang, Wu Jianqiang, Shao Chen, Gao Youhe

机构信息

Department of Pathophysiology, National Key Laboratory of Medical Molecular Biology, Institute of Basic Medical Sciences Chinese Academy of Medical Sciences, School of Basic Medicine Peking Union Medical College, Beijing, 100005, China.

1] Department of Pathophysiology, National Key Laboratory of Medical Molecular Biology, Institute of Basic Medical Sciences Chinese Academy of Medical Sciences, School of Basic Medicine Peking Union Medical College, Beijing, 100005, China [2] Department of Biochemistry and Molecular Biology, Beijing Normal University, Gene Engineering and Biotechnology Beijing Key Laboratory, Beijing, 100875, China.

出版信息

Sci Rep. 2015 Mar 20;5:9314. doi: 10.1038/srep09314.

Abstract

Urine has the potential to become a better source of biomarkers. Urinary proteins are affected by many factors; therefore, differentiating between the variables associated with any particular pathophysiological condition in clinical samples is challenging. To circumvent these problems, simpler systems, such as animal models, should be used to establish a direct relationship between disease progression and urine changes. In this study, a unilateral ureteral obstruction (UUO) model was used to observe tubular injury and the eventual development of renal fibrosis, as well as to identify differential urinary proteins in this process. Urine samples were collected from the residuary ureter linked to the kidney at 1 and 3 weeks after UUO. Five hundred proteins were identified and quantified by LC-MS/MS, out of which 7 and 19 significantly changed in the UUO 1- and 3-week groups, respectively, compared with the sham-operation group. Validation by western blot showed increased levels of Alpha-actinin-1 and Moesin in the UUO 1-week group, indicating that they may serve as candidate biomarkers of renal tubular injury, and significantly increased levels of Vimentin, Annexin A1 and Clusterin in the UUO 3-week group, indicating that they may serve as candidate biomarkers of interstitial fibrosis.

摘要

尿液有潜力成为更好的生物标志物来源。尿蛋白受多种因素影响;因此,区分临床样本中与任何特定病理生理状况相关的变量具有挑战性。为规避这些问题,应使用更简单的系统,如动物模型,来建立疾病进展与尿液变化之间的直接关系。在本研究中,采用单侧输尿管梗阻(UUO)模型观察肾小管损伤及肾纤维化的最终发展情况,并在此过程中鉴定差异尿蛋白。在UUO术后1周和3周,从与肾脏相连的残余输尿管收集尿液样本。通过液相色谱-串联质谱法(LC-MS/MS)鉴定并定量了500种蛋白质,其中与假手术组相比,UUO 1周组和3周组分别有7种和19种蛋白质显著变化。蛋白质印迹法验证显示,UUO 1周组中α-辅肌动蛋白-1和埃兹蛋白水平升高,表明它们可能作为肾小管损伤的候选生物标志物,而UUO 3周组中波形蛋白、膜联蛋白A1和聚集素水平显著升高,表明它们可能作为间质纤维化的候选生物标志物。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/b5ac/4366765/d2dd291d0cb2/srep09314-f1.jpg

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