基于UPLC-Q/TOF MS技术探讨竹节参总皂苷对HepG2细胞脂质代谢的改善作用  

Exploring the Improvement Effect of Total Saponins of Panax Japonicus on Lipid Metabolism in HepG2 Cells Based on UPLC-Q/TOF MS Technology

作　　者：徐睿 钟品菲 周昌园 胡雪黎 袁小鹿 胡泽华 张淇淞 杨宝 XU Rui;ZHONG Pin-fei;ZHOU Chang-yuan;HU Xue-li;YUAN Xiao-lu;HU Ze-hua;ZHANG Qi-song;YANG Bao(Hubei Provincial Key Laboratory of Occurrence and Intervention of Rheumatic Diseases(Hubei Minzu University),Enshi 445000,China;Chinese Medicinal Materials Products Quality Supevision and Inspection Center in Wuling Mountainous Area,Hubei Minzu University,Enshi 445000,China;Medical School,Hubei Minzu University,Enshi 445000,China;School of Pharmaceutical Sciences,Guangzhou University of Chinese Medicine,Guangzhou 510006,China;Medical College of Guangxi University,Nanning 530004,China)

机构地区：[1]风湿性疾病发生与干预湖北省重点实验室(湖北民族大学),湖北恩施445000 [2]湖北民族大学武陵山中药材检验检测中心,湖北恩施445000 [3]湖北民族大学医学部,湖北恩施445000 [4]广州中医药大学中药学院,广东广州510006 [5]广西大学医学院,广西南宁530004

出　　处：《分析测试学报》2023年第11期1434-1441,共8页Journal of Instrumental Analysis

基　　金：国家自然科学基金资助项目(82160804);湖北省自然科学基金资助项目(2021CFB205);风湿性疾病发生与干预湖北省重点实验室(湖北民族大学)项目(PT022202);湖北民族大学大学生创新创业训练计划项目(202210517008)。

摘　　要：基于细胞代谢组学技术研究竹节参总皂苷改善HepG2细胞脂质代谢的机制。应用棕榈酸和油酸诱导HepG2细胞脂质沉积模型,竹节参总皂苷干预24 h后进行油红O染色和甘油三酯分析。采用改良的Bligh-Dy⁃er法将细胞内源性代谢物分为极性和非极性部位,运用超高效液相色谱-四极杆飞行时间质谱(UPLC-Q/TOF MS)技术检测代谢物,结合多元统计分析和火山图筛选差异代谢物,并富集代谢通路。结果显示,竹节参总皂苷可显著改善HepG2细胞脂质沉积模型的脂质代谢。从极性部位鉴定了34个差异代谢物,主要为脂肪酸和氨基酸类成分;从非极性部位鉴定了28个差异代谢物,主要为磷脂酰胆碱、磷脂酰乙醇胺、神经酰胺类成分。差异代谢物在竹节参总皂苷干预后均有恢复至正常水平的趋势。该研究提示竹节参总皂苷改善HepG2细胞脂质沉积模型的脂质代谢与调控鞘脂代谢、甘油磷脂代谢、脂肪酸代谢、炎症反应密切相关。The aim of this study was to investigate the mechanism of the total saponins of Panax japonicus on improving the lipid metabolism in HepG2 cells based on cell metabolomics.The HepG2 lipid deposition model was established with a 1 mmol/L free fatty acid mixture(oleic and palmitic acid at the proportion of 2∶1).Oil red O staining and triglyceride analysis were carried out after 24 h of treatment with the total saponins of Panax japonicus(50 mg/L).Total metabolites in HepG2 cells were extracted according to the improved Bligh-Dyer method and were further separated into polar and nonpolar fractions.Ultra performance liquid chromatography-quadrupole time-of-flight mass spectrometry(UPLC-Q/TOF MS)was used to detect and identify the metabolites.The Progenesis QI software was used to pre-process the data,such as baseline correction,peak alignment,peak recognition,peak extraction and peak normalization.The principal component analysis,orthogonal partial least squares discriminant analysis and volcano map analysis were used to screen the differential metabolites.Metabolic pathways were enriched by the MetaboAnalyst software.The results showed that the total saponins of Panax japonicas could significantly improve the lipid accumulation induced by oleic and palmitic acid in HepG 2 cells.34 differential metabolites were identified in polar fraction,mainly including fatty acids and amino acids.28 differential metabolites were identified from the nonpolar fraction,mainly including phosphatidylcholines,phosphatidyl ethanolamines and ceramides.These differential metabolites tended to return to normal levels after treatment with the total saponins of Panax japonicus.Differential metabolites are mainly involved in three metabolic pathways,including sphingolipid metabolism,glycerol phospholipid metabolism and unsaturated fatty acid synthesis.This study elaborated the possible mechanism of the total saponins of Panax japonicus on ameliorating lipid accumulation induced by oleic and palmitic acid in HepG2 cells from the perspective

关 键 词：竹节参总皂苷 HEPG2细胞 脂质代谢 细胞代谢组学 超高效液相色谱-四极杆飞行时间质谱 

分 类 号：O657.63[理学—分析化学] O657.7[理学—化学]