基于全谱非靶向代谢组学技术的川芎不同部位代谢物深度解析

Differential Analysis of Secondary Metabolism Enrichment in Different Parts of Ligusticum chuanxiong Hort.based on Full Spectrum Non-Targeted Metabolomics

为了深度解析川芎中不同部位代谢物特征,本研究采用高效液相色谱法(HPLC)对川芎的根茎、茎、叶中的活性成分进行含量测定,并整合超高效液相色谱-质谱联用仪(UPLC-MS)和气相色谱-质谱联用仪(GC-MS)的全谱非靶向代谢组学技术,对川芎的根茎、茎和叶的挥发性成分、非挥发性成分进行全面的定性和定量分析。结果表明,川芎根茎中活性成分含量高于茎、叶。川芎的全谱非靶向代谢组学共检出2891个代谢物,总丰度为茎>叶>根茎,各部位化学成分种类相同、含量差异较大,包括氨基酸及其衍生物、萜类、酚酸类等32类化合物。其中LC-MS检出1726个代谢物,GC-MS检出1216个代谢物,两个平台共同检出51个代谢物。对川芎不同部位的差异代谢物进行分析,根茎与茎、根茎与叶、茎与叶中筛选到差异代谢物1683、2054和1844个,差异代谢物总丰度为根茎≈茎>叶。根茎中显著富集含氮化合物、酚类、其他类(糖类、内酯类),茎中的醇、胺类、醚类高度富集,叶中的萜类、酮类、黄酮类高度富集。川芎中大多数活性成分如川芎嗪、阿魏酸、藁本内酯等,呈现根茎>茎>叶的趋势,但茎和叶中也含有含量较高的阿魏酸、欧当归内酯A等重要活性成分,具有较高利用价值。通过KEGG富集分析结果推测差异代谢物可能与次生代谢产物的生物合成等途径相关。本研究深度解析了川芎不同部位的成分积累规律,为川芎资源的合理利用提供了科学依据。

To study the differential characteristics of metabolites in different parts of Ligusticum chuanxiong Hort.and provide reference for the comprehensive utilization of its non-medicinal parts,high performance liquid chromatography(HPLC)was used to determine the content of active ingredients in rhizomes,stems and leaves of L.chuanxiong,the full spectrum non-targeted metabolomics technology of ultra performance liquid chromatography-mass spectrometry(UPLC-MS)and gas chromatography-mass spectrometry(GC-MS)was used to analyze the volatile components and non-volatile metabolites in rhizome,stem and leaf of L.chuanxiong by multivariate statistical non-targeted metabolomics.The results showed that the content of active ingredients in rhizome of L.chuanxiong was higher than that in stem and leaf.A total of 2891 metabolites were detected in L.chuanxiong,and the total abundance was stems>leaves>rhizomes.Although the chemical components of each part were the same,there were significant variations in their contents.These variations included 32 kinds of compounds such as amino acids and their derivatives,terpenes and phenolic acids.A total of 1726 metabolites were detected by LC-MS and 1216 metabolites were detected by GC-MS,among which 51 metabolites were detected in common.The analysis of differential metabolites from different parts of L.chuanxiong revealed 1683,2054 and 1844 differential metabolites between rhizomes and stems,rhizomes and leaves,and stems and leaves,respectively.The total abundance of differential metabolites was rhizomes≈stems>leaves.Nitrogen compounds,phenols and others(Saccharides and Lactones)were significantly enriched in the rhizomes,the ethers,alcohol and amines were highly enriched in the stems,and the terpenoids,ketones and flavonoids in the leaves were highly enriched.Most of the active components of ligustrazine,ferulic acid,ligustilide,etc.are showed the enrichment trend in rhizomes>stems>leaves.Despite the highest concentration of active ingredients being in the rhizomes,the stems and leaves of L.chuanxiong also contained high levels of ferulic acid,levistolide A and other important active ingredients.KEGG enrichment analysis suggested that the differential compositions may be linked to the biosynthesis of secondary metabolites and other pathways.The results comprehensively reflect the accumulation rules and differences of components in different parts of L.chuanxiong,and provide a basis for L.chuanxiong and the rational utilization of above-ground resources.