Udupa Venkatesha, Prakash Veeru
Department of Toxicology, Glenmark Pharmaceuticals Limited, A607, TTC Industrial Area, MIDC, Mahape, Navi Mumbai, 400 709, Maharashtra, India.
Department of Biochemistry and Biochemical Engineering, Jacob Institute of Biotechnology and Bioengineering, Sam Higginbottom University of Agriculture, Technology and Sciences, Allahabad, 211 007, Uttar Pradesh, India.
Toxicol Rep. 2018 Nov 30;6:91-99. doi: 10.1016/j.toxrep.2018.11.015. eCollection 2019.
Consistent, sensitive biomarkers of acute kidney injury in animal models and humans have historically represented a poorly met need for investigators and clinicians. Detection of early kidney damage using urinary biomarkers is essential to assess the adversity in preclinical toxicology studies, which will help in reducing attrition of lead candidates in drug development. This study was undertaken to evaluate recently identified urinary biomarkers use in identifying acute kidney injury compared to traditional serum markers in experimentally induced nephrotoxicity in male Sprague Dawley (SD) rats. Gentamicin induced nephrotoxicity in Sprague Dawley rats is commonly detected using serum markers and histological evaluation of kidneys. Gentamicin, an aminoglycoside was administered at 30 and 100 mg/kg/day dose (subcutaneous) for seven consecutive days to induce nephrotoxicity. On day 4 and day 8 post treatment, serum and urine samples from these rats were analyzed for traditional serum/urine and novel urinary biomarkers and microscopic evaluation of kidneys. On Day 4, no statistically significant change in serum BUN and creatinine level, but increase in urinary microalbumin (mALB) and urinary protein (UP) noticed in both doses of Gentamicin treated rats. On Day 8 significant increase in serum blood urea nitrogen (BUN), serum creatinine, UP and urinary mALB at 100 mg/kg/day, increase in total protein and decrease in albumin in 30 and 100 mg/kg/day and decrease in BUN and creatinine at 100 mg/kg of Gentamicin treated rats. The BUN and creatinine levels or fold change was comparable between control and 30 mg/kg of Gentamicin on Day 8, however, there was 5.6 and 3.4 fold change in BUN and Creatinine level noticed at 100 mg/kg/day of Gentamicin. On Day 4 and 8, significant increase in urinary levels of Clusterin was noted with animals administered both doses of Gentamicin. Similarly, significant increase in urinary levels of kidney injury molecule 1 (Kim-1), Cystatin C and neutrophil gelatinase-associated lipocalin (NGAL) were noticed with animals administered Gentamicin at 100 mg/kg/day on both Day 4 and 8. All these markers have shown dose-dependent change. Histological changes seen on Day 4 and Day 8 were of minimal to mild and moderate to severe in nature at both doses, respectively. The results demonstrated the sensitiveness and accuracy of detecting acute renal damage with novel urinary biomarkers, and their use in diagnosing early kidney damage. This helps in adversity assessment in animal toxicology studies and advocating right treatment to patients who have early renal injury which otherwise can only be diagnosed by elevated levels of traditional biomarkers in blood only after >30% of kidneys is damaged.
在动物模型和人类中,急性肾损伤的一致性、敏感性生物标志物一直以来都远不能满足研究人员和临床医生的需求。在临床前毒理学研究中,使用尿液生物标志物检测早期肾损伤对于评估药物毒性至关重要,这将有助于减少药物开发中候选先导药物的淘汰率。本研究旨在评估最近鉴定出的尿液生物标志物在雄性Sprague Dawley(SD)大鼠实验性诱导的肾毒性中与传统血清标志物相比用于识别急性肾损伤的情况。庆大霉素诱导的SD大鼠肾毒性通常使用血清标志物和肾脏组织学评估来检测。将氨基糖苷类药物庆大霉素以30和100mg/kg/天的剂量(皮下注射)连续给药7天以诱导肾毒性。在治疗后第4天和第8天,对这些大鼠的血清和尿液样本进行传统血清/尿液和新型尿液生物标志物分析以及肾脏显微镜评估。在第4天,庆大霉素处理的两组大鼠血清尿素氮(BUN)和肌酐水平无统计学显著变化,但尿微量白蛋白(mALB)和尿蛋白(UP)增加。在第8天,100mg/kg/天剂量的庆大霉素处理组大鼠血清血尿素氮(BUN)、血清肌酐、UP和尿mALB显著增加,30和100mg/kg/天剂量组大鼠总蛋白增加、白蛋白减少,100mg/kg庆大霉素处理组大鼠BUN和肌酐减少。在第8天,对照组和30mg/kg庆大霉素处理组的BUN和肌酐水平或变化倍数相当,然而,100mg/kg/天庆大霉素处理组的BUN和肌酐水平变化倍数分别为5.6和3.4倍。在第4天和第8天,给予两种剂量庆大霉素的动物尿中Clusterin水平显著增加。同样,在第4天和第8天,给予100mg/kg/天庆大霉素的动物尿中肾损伤分子1(Kim-1)、胱抑素C和中性粒细胞明胶酶相关脂质运载蛋白(NGAL)水平显著增加。所有这些标志物均呈现剂量依赖性变化。在第4天和第8天观察到的组织学变化在两种剂量下分别为轻度至中度和中度至重度。结果证明了新型尿液生物标志物检测急性肾损伤的敏感性和准确性,以及它们在诊断早期肾损伤中的应用。这有助于动物毒理学研究中的毒性评估,并为早期肾损伤患者提供正确的治疗建议,否则只有在超过30%的肾脏受损后才能通过血液中传统生物标志物水平升高来诊断。