Using intracellular metabolic profiling to identify novel biomarkers of cisplatin-induced acute kidney injury in NRK-52E cells
Using intracellular metabolic profiling to identify novel biomarkers of cisplatin-induced acute kidney injury in NRK-52E cells
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DOI：
10.1080/15287394.2021.1969305
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发表时间：
2021-08-29
影响因子：
2.6
通讯作者：
Kim, Hyung Sik
中科院分区：
文献类型：
--
作者：
Kim, Hae Ri;Park, Jae Hyeon;Kim, Hyung Sik
本研究的目的是研究顺铂（CDDP）诱导的正常肾小管上皮细胞NRK-52 E肾毒性引起的细胞内代谢的变化。在用CDDP处理的NRK-52 E细胞中评价细胞毒性、细胞周期分析和凋亡性细胞死亡。随后，使用质子核磁共振（H-1-NMR）光谱来研究细胞代谢谱。在体内模型中测定CDDP诱导的肾毒性。在NRK-52 E细胞中的细胞毒性在用CDDP处理后显著升高，并且发现这些增加是浓度依赖性的。CDDP处理的NRK-52 E细胞中p53和Bax蛋白表达均增加，与细胞凋亡增强相关。此外，在这些细胞中，培养基和细胞裂解物中的许多代谢物都发生了变化。在细胞裂解物中，柠檬酸盐，肌酸酐，和乙酸水平显着降低后，与20 μ M的CDDP浓度的治疗，而谷氨酸水平升高。乳酸盐和乙酸盐水平显着增加培养基中，但柠檬酸盐浓度降低后，高20 μ M CDDP浓度孵育。此外，在CDDP处理的细胞中，丛生蛋白、钙结合蛋白、中性粒细胞明胶酶相关脂质运载蛋白（NGAL）、肾损伤分子-1（KIM-1）、硒结合蛋白1（SBP 1）和丙酮酸激酶M2（PKM 2）向培养基中的分泌显著增加，而乙酰辅酶A合成酶1（AceCS 1）的表达在这些细胞中显著降低。这些结果表明，醋酸依赖性代谢途径可能是一个可靠的和有用的生物标志物检测CDDP诱导的肾毒性。总之，数据表明，通过靶细胞中的代谢产物谱发现新的生物标志物可能有助于检测肾毒性和新药开发。
The aim of this study was to investigate changes in the intracellular metabolism resulting from cisplatin (CDDP)-induced nephrotoxicity in normal kidney tubular epithelial NRK-52E cells. Cytotoxicity, cell cycle analysis, and apoptotic cell death were all evaluated in NRK-52E cells treated with CDDP. Subsequently, proton nuclear magnetic resonance (H-1-NMR) spectroscopy was used to investigate cellular metabolic profiles. CDDP-induced nephrotoxicity was determined in vivo model. Cytotoxicity in the NRK-52E cells significantly rose following treatment with CDDP and these increases were found to be concentration-dependent. Both p53 and Bax protein expression was increased in CDDP-treated NRK-52E cells, correlating with enhanced cellular apoptosis. In addition, a number of metabolites were altered in both media and cell lysates in these cells. In cell lysates, citrate, creatinine, and acetate levels were dramatically reduced following treatment with 20 mu M CDDP concentrations, while glutamate level was elevated. Lactate and acetate levels were significantly increased in culture media but citrate concentrations were reduced following high 20 mu M CDDP concentrations incubation. In addition, excretion of clusterin, calbindin, neutrophil gelatinase-associated lipocalin (NGAL), kidney injury molecule-1 (KIM-1), selenium binding protein 1 (SBP1), and pyruvate kinase M2 (PKM2) into the culture media was significantly increased in CDDP-treated cells while expression of acetyl CoA synthetase 1 (AceCS1) was markedly reduced in these cells. These findings suggest that acetate-dependent metabolic pathway may be a reliable and useful biomarker for detecting CDDP-induced nephrotoxicity. Taken together, data demonstrate that the discovery of novel biomarkers by metabolite profiling in target cells may contribute to the detection of nephrotoxicity and new drug development.