目的 采用¹H NMR的代謝組學(xué)技術(shù)揭示大黃素的腎毒性機(jī)制,尋找腎臟損害的早期生物標(biāo)志物.方法 雄性SD大鼠20只,隨機(jī)分為溶劑對(duì)照,大黃素170、500、1 500 mg/(kg·d)3個(gè)劑量組,連續(xù)給藥16 d,給藥結(jié)束后收集24 h尿液,血漿及腎組織,測(cè)定¹H NMR譜,并進(jìn)行血漿生化指標(biāo)測(cè)定和肝臟組織病理學(xué)檢查.結(jié)果 1 500 mg/(kg·d)大黃素服用16 d可引起大鼠血肌酐下降,大黃素可導(dǎo)致腎細(xì)胞胞漿中出現(xiàn)明顯的空泡化改變.代謝成分的改變主要表現(xiàn)為血液中乳酸、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;腎臟組織中醋酸鹽和肌酐/肌酸明顯升高,乳酸和膽堿/磷酸卵磷脂水平下降,飽和與不飽和脂肪酸及磷脂的成分比例明顯改變.結(jié)論 代謝組學(xué)分析在識(shí)別藥物誘導(dǎo)代謝成分改變方面較傳統(tǒng)技術(shù)更靈敏;脂肪和能量代謝紊亂參與了大黃素的腎毒性,尿液中氨基酸、葡萄糖氧化三甲胺(TMAO)及肌酐可作為大黃素誘導(dǎo)腎組織損害的潛在生物標(biāo)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.

重大新藥創(chuàng)制科技重大專項(xiàng)(2013ZX09302303,2012ZX09301-001-008)