Korea Food and Drug Administration, 5-Nokbun-dong, Eunpyung-gu, Seoul 122-704, Republic of Korea.
Toxicol Appl Pharmacol. 2010 Dec 1;249(2):114-26. doi: 10.1016/j.taap.2010.08.017. Epub 2010 Sep 9.
The primary objective of this study was to determine and characterize surrogate biomarkers that can predict nephrotoxicity induced by mercuric chloride (HgCl₂) using urinary proton nuclear magnetic resonance (¹H NMR) spectral data. A procedure for (1)H NMR urinalysis using pattern recognition was proposed to evaluate nephrotoxicity induced by HgCl₂ in Sprague-Dawley rats. HgCl₂ at 0.1 or 0.75 mg/kg was administered intraperitoneally (i.p.), and urine was collected every 24 h for 6 days. Animals (n=6 per group) were sacrificed 3 or 6 days post-dosing in order to perform clinical blood chemistry tests and histopathologic examinations. Urinary ¹H NMR spectroscopy revealed apparent differential clustering between the control and HgCl₂ treatment groups as evidenced by principal component analysis (PCA) and partial least square (PLS)-discriminant analysis (DA). Time- and dose-dependent separation of HgCl₂-treated animals from controls was observed by PCA of ¹H NMR spectral data. In HgCl₂-treated rats, the concentrations of endogenous urinary metabolites of glucose, acetate, alanine, lactate, succinate, and ethanol were significantly increased, whereas the concentrations of 2-oxoglutarate, allantoin, citrate, formate, taurine, and hippurate were significantly decreased. These endogenous metabolites were selected as putative biomarkers for HgCl₂-induced nephrotoxicity. A dose response was observed in concentrations of lactate, acetate, succinate, and ethanol, where severe disruption of the concentrations of 2-oxoglutarate, citrate, formate, glucose, and taurine was observed at the higher dose (0.75 mg/kg) of HgCl₂. Correlation of urinary (1)H NMR PLS-DA data with renal histopathologic changes suggests that ¹H NMR urinalysis can be used to predict or screen for HgCl₂-induced nephrotoxicity.
本研究的主要目的是利用尿质子磁共振（1H NMR）谱数据确定并表征能够预测氯化汞（HgCl₂）诱导肾毒性的替代生物标志物。提出了一种使用模式识别进行 1H NMR 尿液分析的方法，用于评估 HgCl₂对 Sprague-Dawley 大鼠的肾毒性。以 0.1 或 0.75 mg/kg 的剂量腹腔内（i.p.）给予 HgCl₂，在 6 天内每 24 小时收集尿液一次。在给药后 3 或 6 天处死动物（每组 6 只），以进行临床血液化学测试和组织病理学检查。尿 1H NMR 光谱显示，通过主成分分析（PCA）和偏最小二乘（PLS）判别分析（DA），对照组和 HgCl₂处理组之间存在明显的差异聚类。通过对 1H NMR 光谱数据进行 PCA，观察到 HgCl₂处理动物与对照组的时间和剂量依赖性分离。在 HgCl₂处理的大鼠中，葡萄糖、乙酸盐、丙氨酸、乳酸、琥珀酸盐和乙醇的内源性尿代谢物浓度显著增加，而 2-氧戊二酸、尿囊素、柠檬酸盐、甲酸盐、牛磺酸和 hippurate 的浓度显著降低。这些内源性代谢物被选为 HgCl₂诱导肾毒性的潜在生物标志物。在乳酸、乙酸盐、琥珀酸盐和乙醇的浓度中观察到剂量反应，在较高剂量（0.75 mg/kg）的 HgCl₂中观察到 2-氧戊二酸、柠檬酸盐、甲酸盐、葡萄糖和牛磺酸浓度的严重破坏。尿 1H NMR PLS-DA 数据与肾脏组织病理学变化的相关性表明，1H NMR 尿液分析可用于预测或筛选 HgCl₂诱导的肾毒性。
