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通过聚多巴胺纳米球/适配体纳米复合物与 DNase I 辅助的循环放大偶联,快速灵敏地检测 NGAL 预测急性肾损伤。

Rapid and sensitive detection of NGAL for the prediction of acute kidney injury via a polydopamine nanosphere/aptamer nanocomplex coupled with DNase I-assisted recycling amplification.

机构信息

State Key Laboratory of Natural Medicines, Jiangsu Key Laboratory of TCM Evaluation and Translational Research, Research Center for Traceability and Standardization of TCMs, School of Traditional Chinese Pharmacy, China Pharmaceutical University, Nanjing 211198, P.R. China.

出版信息

Analyst. 2020 May 18;145(10):3620-3625. doi: 10.1039/d0an00474j.

DOI:10.1039/d0an00474j
PMID:32338259
Abstract

Early detection of acute kidney injury (AKI) is important, as early intervention and treatment can prevent further kidney injury and improve kidney health. Neutrophil gelatinase-associated lipocalin (NGAL) has emerged as the earliest and promising non-invasive biomarker of AKI in urine, and has been used as a new predictive biomarker of AKI in the bench-to-bedside journey. In this work, a nanocomplex composed of a polydopamine nanosphere (PDANS) and a fluorophore-labelled aptamer has been constructed for the detection of NGAL using a DNase I-assisted recycling amplification strategy. After the addition of NGAL, the fluorescence intensity increases linearly over the NGAL concentration range from 12.5 to 400 pg mL-1. The limit of detection of this strategy is found to be 6.25 pg mL-1, which is almost 5 times lower than that of the method that does not involve DNase I. The process can be completed within 1 h, indicating a fast fluorescence response. Furthermore, the method using the nanocomplex coupled with DNase I has been successfully utilized for the detection of NGAL in the urine from cisplatin-induced AKI and five-sixths nephrectomized mice, demonstrating its promising ability for the early prediction of AKI. This method also demonstrates the protective effect of the Huangkui capsule on AKI, and provides an effective way to screen potentially protective drugs for renal disease.

摘要

早期发现急性肾损伤 (AKI) 很重要,因为早期干预和治疗可以防止进一步的肾损伤并改善肾脏健康。中性粒细胞明胶酶相关脂质运载蛋白 (NGAL) 已成为尿液中 AKI 的最早和最有前途的非侵入性生物标志物,并已被用作 AKI 从实验室到临床的新预测生物标志物。在这项工作中,构建了一种由聚多巴胺纳米球 (PDANS) 和荧光标记的适体组成的纳米复合物,用于使用 DNase I 辅助循环放大策略检测 NGAL。加入 NGAL 后,荧光强度在 12.5 到 400 pg mL-1 的 NGAL 浓度范围内呈线性增加。发现该策略的检测限为 6.25 pg mL-1,几乎比不涉及 DNase I 的方法低 5 倍。该过程可以在 1 小时内完成,表明具有快速的荧光响应。此外,已成功将使用纳米复合物与 DNase I 结合的方法用于顺铂诱导的 AKI 和六分之五肾切除小鼠尿液中 NGAL 的检测,表明其具有早期预测 AKI 的巨大潜力。该方法还证明了黄葵胶囊对 AKI 的保护作用,并为筛选潜在的肾病保护药物提供了有效的途径。

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