基于代谢组学的麦冬和山麦冬类黄酮化合物的差异性研究

Study on Difference of Flavonoids in Ophiopogonis Radix and Liriopes Radix Based on Metabonomics

关键词:  麦冬  山麦冬  UPLC-MS/MS  代谢组学  类黄酮  KEGG通路

Key words:  Ophiopogonis Radix  Liriopes Radix  UPLC-MS/MS  metabolomics  flavonoids  KEGG pathway

摘要:

目的 研究麦冬和山麦冬药材中类黄酮化合物的分布差异及积累规律,探寻麦冬和山麦冬的特征性差异物质。方法 采用紫外分光光度法和HPLC对麦冬和山麦冬中总黄酮、麦冬甲基黄烷酮A、麦冬甲基黄烷酮B进行定量分析;运用UPLC-MS/MS对麦冬和山麦冬中类黄酮物质进行检测鉴定,利用主成分分析和正交偏最小二乘判别分析法等多元统计分析方法筛选差异代谢物,并对差异代谢物进行KEGG通路富集分析。结果 麦冬中总黄酮、麦冬甲基黄烷酮A,麦冬甲基黄烷酮B的含量显著高于山麦冬。在麦冬和山麦冬中共鉴定出190个类黄酮化合物,其中160个为差异代谢物,主要包括黄酮(31个)、黄酮醇(22个)、异黄酮及其他类黄酮(70个)等。与麦冬比较,山麦冬中有45种物质上调积累,上调物质主要为黄酮和黄酮醇类化合物;有115种物质下调,主要为异黄酮及其他类黄酮化合物。黄酮及黄酮醇类呈现出不同的积累模式,大多数的芹菜素类、山柰酚类在麦冬中上调积累,而橙皮素类、槲皮素类在山麦冬中上调积累。异黄酮及其他类黄酮化合物在麦冬中积累丰富,特别是高异黄酮类在麦冬中全部上调,是麦冬和山麦冬中差异最为明显的一类。通过对160种差异代谢物进行KEGG注释,发现以类黄酮生物合成(ko00941)、异黄酮生物合成(ko00943)、黄酮和黄酮醇生物合成(ko00944) 3条代谢通路富集显著。结论 麦冬和山麦冬中含有丰富的类黄酮物质,整体呈现出明显差异的积累模式,高异黄酮类可作为区分两者的特征成分,该结果可为麦冬类中药品质的深入研究和临床利用提供借鉴。

Abstract:

OBJECTIVE To study the distribution differences and accumulation patterns of flavonoids in Ophiopogonis Radix and Liriopes Radix, and explore the characteristic difference substances between them. METHODS Quantitative analysis of total flavonoids, methylophiopogonanone A and methylophiopogonanone B were carried out by UV-VIS spectrophotometry and HPLC. UPLC-MS/MS was performed to detect the flavonoids in Ophiopogonis Radix and Liriopes Radix. Using multivariate statistical analysis methods such as principal component analysis and orthogonal partial least squares discriminant analysis to identify differential metabolites, and conduct KEGG enrichment analysis of differential metabolites. RESULTS The contents of total flavonoids, methylophiopogonanone A and methylophiopogonanone B in Ophiopogonis Radix were significantly higher than those in Liriopes Radix. A total of 190 flavonoids were identified, of which 160 were differential metabolites, mainly including flavones(31), flavonols(22), isoflavones and other flavonoids(70). Compared with Ophiopogonis Radix, 45 metabolites in Liriopes Radix were up-regulated, and the up-regulated substances were mainly flavones and flavonols; 115 components in Liriopes Radix were down regulated, which were mainly isoflavones and other flavonoids. Flavones and flavonols had different accumulation patterns, most of apigenin and kaempferol were up-regulated in Ophiopogonis Radix, while hesperidin and quercetin were up-regulated in Liriopes Radix. Isoflavones and other flavonoid compounds were abundant in Ophiopogonis Radix, especially homoisoflavonoids were all up-regulated in Ophiopogonis Radix, which was the most significant difference between Ophiopogonis Radix and Liriopes Radix. Through KEGG annotation of 160 differential metabolites, it was found that the differential accumulated flavonoids were significantly enriched in three metabolic pathways:flavonoid biosynthesis(ko00941), isoflavonoid biosynthesis(ko00943), flavone and flavonol biosynthesis(ko00944). CONCLUSION Ophiopogonis Radix and Liriopes Radix both contain rich flavonoids, showing a significantly different accumulation pattern as a whole. Homoisoflavonoids can be considered as the characteristic components to distinguish them, the results can provide reference for the in-depth quality evaluation and clinical utilization of Ophiopogonis Radix and Liriopes Radix.