基于GC-MS的枳壳水煎液抑制THP-1源性巨噬细胞泡沫化代谢组学研究
Metabolomics study of GC-MS-based Aurantii Fructus water decoction inhibition of THP-1-derived macrophage foaming
柯畅 1王燕 1曹国胜 2周仲实 1刘艳菊 2涂济源2
作者信息
- 1. 湖北中医药大学药学院,武汉 430065
- 2. 湖北中医药大学药学院,武汉 430065;湖北省中药炮制技术工程研究中心,武汉 430065
- 折叠
摘要
目的:研究枳壳水煎液(AFD)抑制氧化低密度脂蛋白(ox-LDL)诱导的人单核白血病细胞(THP-1)源性巨噬细胞泡沫化的代谢组学及其可能机制.方法:THP-1细胞分为对照组,模型组,枳壳低、中、高剂量组.采用油红0染色法检测泡沫细胞的形成,用ELISA法检测细胞内甘油三酯(TG)、总胆固醇(TC)及炎症因子水平[单核细胞趋化蛋白1(MCP-1)、干扰素γ(IFN-γ)和肿瘤坏死因子α(TNF-α)],采用GC-MS检测各组细胞代谢组,寻找与药效相关的差异代谢物,并预测相关代谢通路.结果:AFD可抑制泡沫细胞形成,降低TG、TC及炎症因子MCP-1、IFN-γ、TNF-α表达水平(P＜0.01,P＜0.05).代谢组学分析显示,L-丝氨酸、L-苏氨酸、L-甲硫氨酸等7种生物标志物与细胞泡沫化关系密切,给药后显著回调(P＜0.05,P＜0.01).结论:AFD可通过调节甘氨酸/丝氨酸代谢、鞘脂代谢、甲硫氨酸代谢抑制巨噬细胞泡沫化,为动脉粥样硬化的治疗提供依据.
Abstract
Objective:To study the metabolomics and possible mechanism of the water detection of Aurantii Fructus(AFD)inhibiting the foaming of THP-1-derived macrophages induced by ox-LDL.Methods:THP-1 cell was divided into control group,model group,low-dose,middle-dose and high-dose groups.Oil red O staining was used to detect the formation of foam cells,ELISA was used to detect the levels of intracellular triglyceride(TG),total cholesterol(TC)and inflammatory factors[monocyte chemoattractant protein-1(MCP-1),interferon-γ(IFN-γ),tumor necrosis factor-α(TNF-α)],and GC-MS was used to detect the cell metabolome in each group to find the differential metabolites related to drug efficacy,and predict related metabolic pathways.Results:AFD could inhibit foam cell formation and reduce the expression levels of TG,TC,MCP-1,IFN-γ,TNF-α(P＜0.01,P＜0.05).Metabolomics analysis showed that 7 biomarkers including L-serine,L-threonine,and L-methionine were closely related to cell foaming,and were significantly reversed after administration(P＜0.05,P＜0.01).Conclusion:AFD can inhibit cell foaming of macrophages by regulating glycine/serine metabolism,sphingolipid metabolism and methionine metabolism,and provide a basis for the treatment of atherosclerosis.
关键词
枳壳/水煎液/动脉粥样硬化/巨噬细胞泡沫化/代谢组学Key words
Aurantii Fructus/Water detection/Atherosclerosis(AS)/Macrophage foaming/Metabolomics引用本文复制引用
基金项目
湖北中医药大学"青苗计划"资助项目(2021ZZX012)
中药炮制技术传承基地项目(鄂中医通[2022]2号)
出版年
2024
NETL
NSTL
万方数据