基于UPLC-Q-TOF-MS的雷公藤红素的代谢组学研究

UPLC-Q-TOF-MS-based metabolomics study of celastrol

采用质谱技术的代谢学方法系统地对雷公藤红素的代谢开展了研究,筛选了雷公藤红素的代谢产物,并考察雷公藤红素对内源性代谢物水平的影响。通过灌胃给予小鼠雷公藤红素后,收集给药后血、粪和尿样品。采用超高效液相色谱四极杆飞行时间质谱仪联用技术(UPLC-Q-TOF-MS)对雷公藤红素的代谢产物及其影响的内源性代谢物进行分析。运用质量亏损过滤策略筛选雷公藤红素的代谢产物,运用多元变量统计分析确定它显著影响的内源性代谢物。结果显示,在小鼠的粪便和尿液中检测到雷公藤红素的原型和8个代谢产物,其中5个代谢产物为首次报道;在鼠肝微粒体和人肝微粒代谢中发现雷公藤红素可以转化生成2个羟化产物,进一步的重组酶孵育揭示CYP3A4是催化其形成羟化产物的主要代谢酶。尿液代谢组学揭示雷公藤红素可以影响hippuric acid,phenylacetylglycine,5-hydroxyindoleacetic acid,urocanic acid,cinnamoylglycine,phenylprop-lonylglycine,xanthurenic acid的排泄,这些代谢物与肠道菌代谢密切相关。上述研究结果有助于阐明雷公藤红素在体内的代谢处置过程及其潜在的药理作用机制。

The mass spectrometry-based metabolomics method was used to systematically investigate the formation of celastrol me-tabolites,and the effect of celastrol on endogenous metabolites.The mice plasma,urine and feces samples were collected after oral ad-ministration of celastrol.Ultra-high performance liquid chromatography with quadrupole time-of-flight mass spectrometry(UPLC-Q-TOF-MS)was applied to analyze the exogenous metabolites of celastrol and its altered endogenous metabolites.Mass defect filtering was adopted to screen for the exogenous metabolites of celastrol.Multivariate statistical analysis was used to identify the endogenous metabo-lites affected by celastrol.Celastrol and its eight metabolites were detected in urine and feces of mice,and 5 metabolites of them were reported for the first time.The hydroxylated metabolites were observed in the metabolism of both human liver microsomes and mouse liv-er microsomes.Further recombinant enzyme experiments revealed CYP3A4 was the major metabolic enzyme involved in the formation of hydroxylated metabolites.Urinary metabolomics revealed that celastrol can affect the excretion of intestinal bacteria-related endogenous metabolites,including hippuric acid,phenylacetylglycine,5-hydroxyindoleacetic acid,urocanic acid,cinnamoylglycine,phenylproplo-nylglycine and xanthurenic acid.These results are helpful to elucidate the metabolism and disposition of celastrol in vivo,and its mech-anism of action.