目的 采用¹H NMR的代谢组学技术揭示大黄素的肾毒性机制,寻找肾脏损害的早期生物标志物.方法 雄性SD大鼠20只,随机分为溶剂对照,大黄素170、500、1 500 mg/(kg·d)3个剂量组,连续给药16 d,给药结束后收集24 h尿液,血浆及肾组织,测定¹H NMR谱,并进行血浆生化指标测定和肝脏组织病理学检查.结果 1 500 mg/(kg·d)大黄素服用16 d可引起大鼠血肌酐下降,大黄素可导致肾细胞胞浆中出现明显的空泡化改变.代谢成分的改变主要表现为血液中乳酸、糖、氨基酸和脂肪酸成分下降;尿液中乳酸、糖和氨基酸成分增加;肾脏组织中醋酸盐和肌酐/肌酸明显升高,乳酸和胆碱/磷酸卵磷脂水平下降,饱和与不饱和脂肪酸及磷脂的成分比例明显改变.结论 代谢组学分析在识别药物诱导代谢成分改变方面较传统技术更灵敏;脂肪和能量代谢紊乱参与了大黄素的肾毒性,尿液中氨基酸、葡萄糖氧化三甲胺(TMAO)及肌酐可作为大黄素诱导肾组织损害的潜在生物标志物.

Objective To reveal the mechanism of nephrotoxicity of emodin using ¹H-NMR spectroscopy and to screen the potential biomarker of renal injury. Methods In this study, the biochemical constituents of urine, serum, and kidney tissue extracts of rats treated with emodin at different doses (0, 170, 500, and 1, 500 mg/kg/d for 16d, i.g.)were investigated using ¹H-NMR techniques and pattern recognition method (PCA/PLS). Serum biochemical analysis and histopathological examination of kidney of all rats were simultaneously performed. Results Emodin (1 500 mg/kg/d) induced slight renal proximal tubule vacuolar degeneration in rats treated 16 d, while serum biochemical indexes were not affected. ¹H-NMR analysis found that the levels of lactate, glucose, many kinds of amino acids, and fat acid decreased in serum. Elevated creatinine, trimetlylamine oxide (TMAO), acetate, lactate, glucose, many kinds of amino acids and decreased citrate, hippurate, and 2-oxoglutarate were found in urine. The predominant changes identified in kidney tissue involved the reduced levels of lactate and choline/phosphatidylcholine. Additionally, the alteration was noted between the proportion of the saturated and unsaturated fatty acid and phospholipid composition. Conclusion The data generated from the current study supports the fact that ¹H-NMR technology is more sensitive than traditional urine and serum analysis. Glucose, amino acids, TMAO, and creatinine in urine could be identified as biomarkers of emodin induced renal injury.

重大新药创制科技重大专项(2013ZX09302303,2012ZX09301-001-008)