藏族药二十五味松石丸治疗慢性胆汁淤积小鼠的血清代谢组学研究

Serum metabolomics of Tibetan medicine Ershiwuwei Songshi Pills against chronic cholestasis in mice

采用喂养含3,5-二乙氧基羰基-1,4-二氢-2,4,6-三甲基吡啶(3,5-diethoxy carbonyl-1,4-dihydro-2,4,6-trimethylpyridine,DDC)的饲料建立慢性胆汁淤积小鼠模型,基于液相色谱-质谱(LC-MS)检测技术的代谢组学分析方法,探究二十五味松石丸(Ershiwuwei Songshi Pills,ESP)对慢性胆汁淤积小鼠的内源性代谢物的影响。结果发现ESP能够有效改善模型组小鼠的病理性损伤以及降低血清丙氨酸氨基转移酶(ALT)、天冬氨酸氨基转移酶(AST)、碱性磷酸酶(ALP)、总胆汁酸(TBA)水平。代谢组学筛查得到模型组与空白组、模型组与ESP组13个共有的差异代谢物,分别是尿酸(uric acid)、乙醇醛(glycolaldehyde)、犬尿氨酸(kynurenine)、黄素腺嘌呤二核苷酸(flavin adenine dinucleotide)、L-3-苯乳酸(L-3-phenyllactic acid)、I-尿胆素(I-urobilin)、白三烯D4(LTD4)、牛磺胆酸(taurocholic acid)、trioxilin A3、1D-肌醇1,4-二磷酸(D-inositol-1,4-diphosphate)、PC[16:0/20:2(11Z,14Z)]、PC[14:0/22:2(13Z,16Z)]、PC[20:4(5Z,8Z,11Z,14Z)/20:4(5Z,8Z,11Z,14Z)]。ESP干预后,13个差异代谢物的含量均有明显回调,通路分析显示,ESP主要通过影响花生四烯酸代谢、甘油磷脂代谢、色氨酸代谢、初级胆汁酸的生物合成等途径来达到治疗效果。该研究从代谢物角度阐释了二十五味松石丸抗慢性胆汁淤积的作用机制。

A chronic cholestasis model was induced in mice by feeding a diet containing 3,5-diethoxycarbonyl-1,4-dihydro-2,4,6-trimethylpyridine(DDC).The effects of Ershiwuwei Songshi Pills(ESP)on endogenous metabolites in mice with chronic cholestasis were investigated by metabolomics analysis based on liquid chromatography-mass spectrometry(LC-MS).The results showed that ESP was effective in improving pathological injury and reducing serum levels of alanine aminotransferase(ALT),aspartate aminotransferase(AST),alkaline phosphatase(ALP),and total bile acid in the model mice.Meanwhile,13 common differential metabolites were revealed in metabolomic screening between the model/control group and the model/ESP group,including uric acid,glycolaldehyde,kynurenine,flavin adenine dinucleotide,L-3-phenyllactic acid,I-urobilin,leukotriene D4(LTD4),taurocholic acid,trioxilin A3,D-inositol-1,4-diphosphate,PC[16:0/20:2(11Z,14Z)],PC[14:0/22:2(13Z,16Z)],and PC[20:4(5Z,8Z,11Z,14Z)/20:4(5Z,8Z,11Z,14Z)].After ESP intervention,the levels of all 13 differential metabolites were significantly retraced,and pathway analysis showed that ESP achieved its therapeutic effect mainly by affecting arachidonic acid metabolism,glycerophospholipid metabolism,tryptophan metabolism,and primary bile acid biosynthesis.This study elucidated the mechanism of action of ESP against chronic cholestasis based on metabolites.