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尿肾损伤分子-1作为梗阻性急性肾损伤的早期诊断生物标志物及快速检测方法的研发

Urinary kidney injury molecule‑1 as an early diagnostic biomarker of obstructive acute kidney injury and development of a rapid detection method.

作者信息

Jin Yingli, Shao Xiaona, Sun Bo, Miao Chunsheng, Li Zhengqiang, Shi Yan

机构信息

Department of Pharmacology, College of Basic Medical Science, Jilin University, Changchun, Jilin 130021, P.R. China.

Department of Experimental Pharmacology and Toxicology, School of Pharmacy, Jilin University, Changchun, Jilin 130021, P.R. China.

出版信息

Mol Med Rep. 2017 Mar;15(3):1229-1235. doi: 10.3892/mmr.2017.6103. Epub 2017 Jan 5.

Abstract

The aim of the present study was to investigate whether urinary kidney injury molecule‑1 (KIM‑1) presents a suitable early diagnostic biomarker of obstructive nephropathy‑induced acute kidney injury (AKI), and to develop a rapid detection method for urinary KIM‑1. Obstructive AKI was induced in an experimental rat model by a unilateral ureteral obstruction (UUO) operation. Macro‑ and micromorphological kidney alterations were determined by visual observation and hematoxylin and eosin (HE) staining, respectively. Kidney functions were evaluated by detecting urea nitrogen and creatinine levels in rat urine and blood. Urinary KIM‑1 levels were measured using an enzyme‑linked immunosorbent assay, and the protein expression levels of KIM‑1, α‑smooth muscle actin (α‑SMA) and vimentin in kidney tissues were detected using immunohistochemical assays. In order to measure KIM‑1 levels, colloidal gold immunochromatographic strips were developed based on the colloidal gold immunochromatographic assay. The results indicated that KIM‑1 levels were significantly higher in the UUO group when compared with the Sham group. KIM‑1 levels in the urine and kidney tissues exhibited a time‑dependent increase, together with increasing obstructive AKI in the UUO group. In addition, KIM‑1 levels were demonstrated to be a more sensitive biomarker of early obstructive AKI, when compared with α‑SMA and vimentin. A colloidal gold‑based immunochromatographic strip was developed, whereby the detection of urinary KIM‑1 could be completed within 5‑10 min. In conclusion, results of the present study demonstrated that urinary KIM‑1 may be a valuable biomarker for the early diagnosis of obstructive AKI, and the use of a colloidal gold immunochromatographic strip may be a promising method for the rapid detection of urinary KIM‑1.

摘要

本研究的目的是探讨尿肾损伤分子-1(KIM-1)是否是梗阻性肾病所致急性肾损伤(AKI)的合适早期诊断生物标志物,并开发一种尿KIM-1的快速检测方法。通过单侧输尿管梗阻(UUO)手术在实验大鼠模型中诱导梗阻性AKI。分别通过肉眼观察和苏木精-伊红(HE)染色确定肾脏的大体和微观形态学改变。通过检测大鼠尿液和血液中的尿素氮和肌酐水平来评估肾功能。使用酶联免疫吸附测定法测量尿KIM-1水平,并使用免疫组织化学测定法检测肾组织中KIM-1、α-平滑肌肌动蛋白(α-SMA)和波形蛋白的蛋白表达水平。为了测量KIM-1水平,基于胶体金免疫层析法开发了胶体金免疫层析试纸条。结果表明,与假手术组相比,UUO组的KIM-1水平显著更高。UUO组尿液和肾组织中的KIM-1水平呈现出时间依赖性升高,同时梗阻性AKI也在加重。此外,与α-SMA和波形蛋白相比,KIM-1水平被证明是早期梗阻性AKI更敏感的生物标志物。开发了一种基于胶体金的免疫层析试纸条,可在5-10分钟内完成尿KIM-1的检测。总之,本研究结果表明,尿KIM-1可能是梗阻性AKI早期诊断的有价值生物标志物,使用胶体金免疫层析试纸条可能是快速检测尿KIM-1的一种有前景的方法。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/f446/5367370/7fae28d80ac7/MMR-15-03-1229-g00.jpg

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