骨碎补醇提物对骨质疏松预防作用的大鼠尿液UPLC-MS/MS代谢组学研究

目的:研究糖皮质激素性骨质疏松大鼠尿液的UPLC-MS/MS代谢物谱变化以及骨碎补醇提物对其的干预作用,探讨液质联用技术在中药代谢组学研究中的应用。方法:采用超高效液相色谱串联质谱(UPLC-MS/MS)技术对正常对照组、骨质疏松模型组、骨碎补给药组大鼠尿液代谢物谱进行分析测定,并对数据进行主成分分析(PCA)以鉴别尿液中代谢产物的变化。结果:正常对照组、骨质疏松模型组、骨碎补给药组得到明显分离,发现了肌酸酐、色氨酸、苯丙氨酸、甲酚硫酸盐、柠檬酸、壬二酸等9个潜在的生物标志物。与正常对照组相比,模型组大鼠尿液中肌酸酐、柠檬酸、壬二酸、马尿酸、色氨酸以及吲哚硫酸盐含量显著下调;苯丙氨酸、甲酚硫酸盐及苯乙酰甘氨酸含量显著上调。结论:骨碎补醇提物可以调节骨质疏松大鼠体内的代谢紊乱,其作用机制可能与调节能量代谢、肠道菌代谢、氨基酸代谢以及抗氧化损伤有关。

Metabonomic study on protective effect of ethanol extracts of Drynariae Rhizoma on osteoporosis in rats urine by using UPLC-MS/MS

This paper was designed to study metabonomic characters of the osteoporosis induced by high dose of hydrocortisone and the protective effects of Drynariae Rhizoma, which can replenish the kidney and strengthen the bones. A urinary metabonomics method based on ultra-performance liquid chromatography coupled with mass spectrometry (UPLC-MS/MS) was developed. Clear separation of healthy control group, model group and treatment group was achieved by using the principal components analysis (PCA) and 9 significantly changed metabolites were identified as potential biomarkers of osteoporosis. Compared with the health control group,the model group rats showed lower levels of creatinine, citric acid, azelaic acid, hippurate, tryptophan and indoxyl sulfate together with higher levels of phenylalanine, cresol sulfate and phenaceturic acid. These changes in urinary metabolites suggest that the disorders of amino acid metabolism, energy metabolism, gut microflora and anti-oxidative damage are related to osteoporosis induced by high dose of hydrocortisone and the potential effect of Drynariae Rhizoma on all the four metabolic pathways.