基于UPLC-Q-Exactive Orbitrap-MS技术分析炙淫羊藿炮制前后对细胞内源性代谢物的影响

Analysis of effect of Epimedium on cells endogenous metabolites before and after the processing based on UPLC-Q-Exactive Orbitrap-MS

目的基于细胞代谢组学分析炙淫羊藿炮制前后对小鼠黑色素瘤细胞(B16细胞)内源性代谢物的影响,探讨炙淫羊藿炮制前后的药性变化。方法采用超高效液相色谱串联四级杆轨道阱质谱(UHPLC-Q-Exactive Orbitrap-MS)技术,对淫羊藿、炙淫羊藿干预的B16细胞内源性小分子进行代谢组学分析,获取组间差异代谢产物,并基于MetaboAnalyst 5.0数据库分析相关代谢通路。结果经炙淫羊藿及其生品干预后,B16细胞中13个细胞内源性差异性代谢物发生变化,分别为丙氨酸、肉毒碱C3∶0、谷氨酸-1、乳酸、异亮氨酸、胆碱、磷脂酰胆碱(34∶2、36∶2)、游离脂肪酸、柠檬酸、肉毒碱C4∶0、溶血磷脂酰胆碱16∶0、苹果酸,且炮制品对差异代谢物的影响强于生品。涉及的主要通路有瓦博格效应、丙酮酸代谢、苹果酸-天冬氨酸穿梭、丙酮醛降解等。结论淫羊藿、炙淫羊藿对细胞代谢途径有一定影响,可能与淫羊藿炮制前后药效作用、寒热药性变化有关。

Objective To analyze the effects of processed Epimedii Folium on endogenous metabolites of mouse melanoma cells(B16 cells)before and after processing based on cell metabolomics;To investigate the changes of processed Epimedii Folium before and after processing.Methods Ultra performance liquid chromatography tandem four-stage orbital trap mass spectrometry(UPLC-Q-Exactive Orbitrap-MS)technology was used,and the endogenous small molecules of B16 cells treated with Epimedii Folium and processed Epimedii Folium were analyzed by metabolomics.The differential metabolites between groups were obtained,and relevant metabolic pathways were analyzed based on the MetaboAnalyst 5.0 database.Results Significant changes were observed in 13 kinds of endogenous metabolites,including alanine,carnitine C3∶0,glutamic acid-1,lactic acid,isoleucine,choline,phosphatidylcholine(34∶2,36∶2),free fatty acids,citric acid,carnitine C4∶0,lysophosphatidylcholine 16∶0 and malic acid after the intervention of Epimedii Folium and processed Epimedii Folium.And the impact of processed products on differential metabolites was stronger than that of raw products.The main pathways involved were Warburg effect,pyruvate metabolism,malate-aspartic acid shuttle,pyruvaldehyde degradation and so on.Conclusions Epimedii Folium and processed Epimedii Folium would have certain effects on cellular metabolic pathways.The results may be related to the pharmacological effects and changes in cold and hot properties of Epimedii Folium before and after processing.