a Analysis Research Team, R&D Center, GL Pharm Tech Corp ., Gyeonggi-do , Republic of Korea.
b Department of Pharmacy, College of Pharmacy, Dankook University , Chungnam , Republic of Korea.
J Toxicol Environ Health A. 2018;81(11):408-420. doi: 10.1080/15287394.2018.1451180. Epub 2018 Apr 2.
Gentamicin (GM) is an aminoglycoside antibiotic used in treatment of various types of bacterial infections, but the major adverse effect is drug-induced nephrotoxicity. This study aimed to determine biomarkers that might predict nephrotoxicity initiated by GM using serum or urinary proton nuclear magnetic resonance (H NMR) spectral data in male Sprague-Dawley rats. GM (0, 30, or 300 mg/kg/d) was intraperitoneally administered for 3 consecutive days. Animals were sacrificed 2 d (D2) or 8 d (D8) after last administration of GM in order to perform analysis of serum biochemistries and histopathologic examination. Urine samples were collected every 24 h from prior to treatment until sacrifice. Serum and urinary H NMR spectral data revealed apparent differential clustering between control and GM-treated groups as evidenced by principal component analysis (PCA) and orthogonal projections to latent structure-discriminant analysis (OPLS-DA) in global and targeted profiling. The concentrations of endogenous serum metabolites including 3-hydroxybutyrate, alanine, citrate, creatine, glucose, and glycine were increased significantly on D2 or D8. Urinary levels of glucose, glycine, and succinate were significantly elevated on D2 or D8, whereas the concentration of hippurate was significantly decreased on D2 and D8. Correlation of serum and urinary H NMR OPLS-DA with serum biochemistry and renal histopathologic changes suggests that H NMR urinalysis may be used to reliably predict or screen for GM-induced nephrotoxicity. In contrast, Western blot analysis of kidney injury molecule-1 (KIM-1) demonstrated that protein expression was not markedly altered indicating this biomarker was not sensitive to detect GM-mediated renal damage. Data suggest that these altered metabolites might serve as specific and sensitive biomarkers for GM-mediated renal damage.
庆大霉素（GM）是一种氨基糖苷类抗生素，用于治疗各种类型的细菌感染，但主要的不良反应是药物引起的肾毒性。本研究旨在使用雄性 Sprague-Dawley 大鼠的血清或尿液质子磁共振（H NMR）光谱数据，确定可能预测 GM 引发的肾毒性的生物标志物。GM（0、30 或 300mg/kg/d）连续腹腔注射 3 天。动物在 GM 最后一次给药后 2 天（D2）或 8 天（D8）处死，以便进行血清生化分析和组织病理学检查。在治疗前至处死期间，每隔 24 小时收集尿液样本。血清和尿液 H NMR 光谱数据显示，通过主成分分析（PCA）和正交偏最小二乘判别分析（OPLS-DA）在全局和靶向分析中，对照组和 GM 处理组之间存在明显的聚类差异。血清中包括 3-羟基丁酸、丙氨酸、柠檬酸、肌酸、葡萄糖和甘氨酸在内的内源性代谢物浓度在 D2 或 D8 时显著升高。尿液中葡萄糖、甘氨酸和琥珀酸水平在 D2 或 D8 时显著升高，而 hippurate 浓度在 D2 和 D8 时显著降低。血清和尿液 H NMR OPLS-DA 与血清生化和肾脏组织病理学变化的相关性表明，H NMR 尿液分析可用于可靠地预测或筛选 GM 引起的肾毒性。相比之下，肾脏损伤分子-1（KIM-1）的 Western blot 分析表明，蛋白质表达没有明显改变，表明该生物标志物对检测 GM 介导的肾损伤不敏感。数据表明，这些改变的代谢物可能作为 GM 介导的肾损伤的特异性和敏感生物标志物。
