基于UPLC-MS/MS的短柄龙胆花和叶代谢组学分析

UPLC-MS/MS-based Metabonomic Analysis between Gentiana stipitata Edgew.Flowers and Leaves

目的为深入研究短柄龙胆花和叶两种组织中的差异代谢物和代谢途径,开展广泛靶向代谢组学研究,为短柄龙胆药用部位的有效利用提供理论依据。方法使用超高效液相色谱串联质谱(UPLC-MS/MS)技术,通过主成分分析(PCA)、正交偏最小二乘判别分析(OPLS-DA)以及聚类分析(HCA)等多元统计学方法寻找短柄龙胆花和叶之间的不同代谢物及代谢途径。结果在短柄龙胆花和叶中共检测到518种代谢物,PCA结果显示短柄龙胆花和叶中的代谢物存在差异,所有代谢物中有336种差异代谢物含量变化显著(P<0.05,VIP≥1),其中倍萜、二氢黄酮醇、黄酮、花青素、木脂素、酚酸、异黄酮、黄酮醇、黄烷醇、黄酮碳糖苷、三萜、环烯醚萜、香豆素、鞣质及其它类黄酮上调和下调幅度均在前20。KEGG中共得到33条差异代谢途径,酪氨酸代谢、黄酮类化合物生物合成、次级代谢物生物合成、黄酮和黄酮醇生物合成和异黄酮类化合物生物合成为5条差异代谢物显著富集(P<0.5)途径。结论本研究通过代谢组学技术揭示了短柄龙胆花和叶中的主要差异代谢物为黄酮类化合物,黄酮类化合物生物合成为显著差异代谢途径。

OBJECTIVE To investigate in depth the differential metabolites and metabolic pathways in both flower and leaf tissues of Gentiana stipitata Edgew.,and to carry out extensive targeted metabolomics studies to provide a theoretical basis for the effective utilization of the medicinal parts of G.stipitata.METHODS The different metabolites and metabolic pathways between G.stipitata flowers and leaves were searched by multivariate statistical methods such as principal component analysis(PCA),orthogonal partial least squares discriminant analysis(OPLS-DA)and cluster analysis(HCA)using ultra performance liquid chromatography tandem mass spectrometry(UPLC-MS/MS)techniques.RESULTS A total of 518 metabolites were detected in G.stipitata flowers and leaves.And PCA results showed that there were differences in metabolites between G.stipitata flowers and leaves,with 336 different metabolites varying significantly in content among all metabolites(P<0.05,VIP≥1),including terpenes,dihydroflavonols,flavonoids,anthocyanins,lignans,phenolic acids,isoflavones,flavonols,flavonoid carbosinosides,triterpenoids,cycloalkenyl ether terpene,coumarins,tannin and other flavonoids up-regulated and down-regulated in the top 20.33 differential metabolic pathways were obtained in KEGG,and tyrosine metabolism,flavonoid biosynthesis,secondary metabolite biosynthesis,flavonoid and flavonol biosynthesis and isoflavonoid biosynthesis were the five differential metabolic significantly enriched pathways(P<O.5).CONCLUSION The present study reveales that the major differential metabolites in G.stipitata flowers and leaves are flavonoids and flavonoid biosynthesis as significantly differential metabolic pathways by metabolomics techniques.