大鼠急性肾损伤模型中15种新型尿液生物标志物的评价研究  

Evaluation of 15 novel urine biomarkers in an animal model of acute kidney injury induced by gentamicin

作　　者：朱思睿 屈哲[1] 杨艳伟[1] 刘鑫磊 黄芝瑛[2] 周晓冰[1] 李波[1] ZHU Si-rui;QU Zhe;YANG Yan-wei;LIU Xin-lei;HUANG Zhi-ying;ZHOU Xiao-bing;LI Bo(National Center for Safety Evaluation of Drugs,National Institutes for Food and Drug Control,Beijing Key Laboratory of Drug Nonclinical Safety Evaluation Research,Beijing 100176,China;School of Pharmaceutical Sciences,Sun Yat-sen University,Guangzhou 510006,China)

机构地区：[1]中国食品药品检定研究院国家药物安全评价监测中心,药物非临床安全评价研究北京市重点实验室,北京100176 [2]中山大学药学院,广州510006

出　　处：《中国医药生物技术》2022年第4期313-320,共8页Chinese Medicinal Biotechnology

基　　金：国家自然科学基金(81603210);“重大新药创制”国家科技重大专项(2018ZX09201017-001)。

摘　　要：目的 评价短期采尿下15种新型尿液生物标志物对药物肾毒性的诊断效能。方法 Wistar大鼠连续腹腔注射120mg/kg庆大霉素构建急性肾损伤(AKI)模型。分别在检疫期、给药第5、8天采集动物5h尿液和血样,并解剖动物,进行肾脏病理学检查。全自动生化分析仪检测各组动物的血清和尿生化。应用Luminex液相芯片方法检测分析尿液生物标志物浓度,与传统肾功能指标尿素氮(BUN)和肌酐(CRE)进行比较。结果 组织病理学结果显示,给药动物出现典型急性肾损伤变化,并呈明显时间相关性;与对照组比较,干扰素诱导蛋白-10(IP-10)和β2-微球蛋白(β2-MG)在给药第5天就显著升高,且持续增加。在给药第8天,除表皮生长因子(EGF)、骨桥蛋白(OPN)两种生物标志物以外,尿液中其他8种生物标志物浓度均明显升高(P <0.01)。而BUN和CRE也在给药第8天出现显著升高(P <0.05),受试者工作特性曲线(ROC)表明,白蛋白(ALB)、尿蛋白(uTP)、肾损伤因子-1(KIM-1)、IP-10、β2-MG、中性粒细胞明胶酶相关脂质运载蛋白(NGAL)、丛生蛋白(CLU)、半胱氨酸蛋白酶抑制剂-C(CysC)和酸性糖蛋白(AGP)的曲线下面积明显优于BUN和CRE。结论 证实ALB、uTP、KIM-1、IP-10、β2-MG、NGAL对药物致急性肾毒性早期诊断效能优于BUN和CRE,这些候选生物标志物在药物的大鼠临床前安全性研究中将具有良好的应用前景。Objective To evaluate the diagnostic efficacy of 15 novel urine biomarkers for drug nephrotoxicity.Methods Wistar rats were given continuous intraperitoneal injection of 120 mg/kg gentamicin(GEN) to construct an acute kidney injury(AKI) model. The 5-hour urine and blood samples were collected during the quarantine period and on the 5th and 8th day after administration, and the animals were dissected for renal pathological examination. An automatic biochemical analyzer was used for serum and urine biochemical testing in each group of animals. The Luminex liquid chip method was used to analyze the concentration of urine biomarkers as compared with the traditional renal function indicators of urea nitrogen(BUN) and creatinine(CRE).Results The histopathological results showed that typical acute kidney injury changes were found in gentamicin-treated animals as an obviously time-related manner. Compared with the control group, interferon-inducible protein-10(IP-10) and β2-microglobulin(β2-MG) were significantly up on the 5th day, and continued to increase. On the 8th day, in addition to the two biomarkers of epidermal growth factor(EGF) and osteopontin(OPN), other 8 biomarkers in urine increased significantly(P < 0.01). BUN and CRE also increased significantly on the 8th day(P < 0.05). The receiver operating characteristic curve(ROC) results showed that the area under the curve(AUC) of albumin(ALB), IP-10, kidney injury factor-1(KIM-1), neutrophil gelatinase-associated lipocalin(NGAL)and β2-MG were significantly better than BUN and CRE.Conclusion The 5-hour urine samples from the drug-induced AKI animal model are collected and analyzed, of which confirmed that ALB, IP-10, KIM-1, NGAL and β2-MG are better than BUN and CRE in the early diagnosis of drug-induced acute nephrotoxicity. Candidate biomarkers will have good application prospects in the preclinical safety studies of drugs in rats.

关 键 词：肾毒性 生物标志物 急性肾损伤 庆大霉素 临床前安全性评价 

分 类 号：R692[医药卫生—泌尿科学]