There have been intensive efforts to identify in vivo biomarkers that can be used to monitor drug-induced kidney damage before significant impairment occurs. Kidney injury molecule-1, neutrophil gelatinase-associated lipocalin, clusterin, β2-microglobulin and cystatin C (CysC) have been validated as clinical or preclinical biomarkers in urinary and plasma predictive of acute and chronic kidney injuries and diseases. A high-throughput in vitro assay predictive of nephrotoxicity could potentially be implemented in early drug discovery stage to reduce attrition at later stages of drug development. To assess the potential of these known in vivo biomarkers for in vitro evaluation of drug-induced nephrotoxicity, we selected four nephrotoxic agents (cisplatin, cyclosporin, aristolochic acid I and gentamicin) and detected their effects on the protein levels of nephrotoxic biomarkers in RPTEC/TERT1 cells. The protein levels of clusterin, CysC, GSTπ and TIMP-1 significantly increased in the conditioned media of RPTEC/TERT1 cells treated with cisplatin, cyclosporin, aristolochic acid I and gentamicin. The messenger RNA levels of clusterin, CysC, GSTπ and TIMP-1 also increased in RPTEC/TERT1 cells treated with cisplatin, cyclosporin, aristolochic acid I and gentamicin, indicating that drug-induced upregulation involves transcriptional activation. Taken together, the results clearly demonstrate that among the known in vivo nephrotoxic biomarkers, clusterin, CysC, GSTπ and TIMP-1 can be effectively used as in vitro biomarkers for drug-induced nephrotoxicity in RPTEC/TERT1 cells.

中文翻译：

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在体外预测RPTEC / TERT1细胞中药物诱导的肾毒性的生物标记物的评估。

已经进行了广泛的努力以鉴定可用于在显着损害发生之前监测药物诱导的肾损伤的体内生物标志物。肾脏损伤分子-1，与中性粒细胞明胶酶相关的脂质运载蛋白，簇蛋白，β2-微球蛋白和胱抑素C（CysC）已被证实在尿液和血浆中可作为临床或临床前生物标志物，预测急性和慢性肾脏损伤和疾病。预测肾毒性的高通量体外测定法可能会在药物开发的早期阶段实施，以减少药物开发后期的磨损。为了评估这些已知的体内生物标记物在体外评估药物诱导的肾毒性的潜力，我们选择了四种肾毒性剂（顺铂，环孢菌素，并检测它们对RPTEC / TERT1细胞中肾毒性生物标志物蛋白质水平的影响。在用顺铂，环孢菌素，马兜铃酸I和庆大霉素处理的RPTEC / TERT1细胞的条件培养基中，簇蛋白，CysC，GSTπ和TIMP-1的蛋白质水平显着增加。在用顺铂，环孢菌素，马兜铃酸I和庆大霉素处理的RPTEC / TERT1细胞中，簇蛋白，CysC，GSTπ和TIMP-1的信使RNA水平也增加，表明药物诱导的上调涉及转录激活。两者合计，结果清楚地表明，在已知的体内肾毒性生物标志物中，簇蛋白，CysC，GSTπ和TIMP-1可以有效地用作体外生物标志物，以用于RPTEC / TERT1细胞中药物诱导的肾毒性。