基于花生四烯酸代谢通路研究水飞蓟素对脂多糖诱导炎症模型影响

引用本文: 麦旦提, 杨婵, 薛芸, 王彦, 阎超. 基于花生四烯酸代谢通路研究水飞蓟素对脂多糖诱导炎症模型影响[J]. 色谱, 2017, 35(6): 578-586. doi: 10.3724/SP.J.1123.2017.01026 shu

Citation: MAI Danti, YANG Chan, XUE Yun, WANG Yan, YAN Chao. Investigation on silymarin impact on lipopolysaccharide induced inflammation model based on arachidonic acid metabolism pathway[J]. Chinese Journal of Chromatography, 2017, 35(6): 578-586. doi: 10.3724/SP.J.1123.2017.01026 shu

基于花生四烯酸代谢通路研究水飞蓟素对脂多糖诱导炎症模型影响

上海交通大学药学院, 上海 200210
基金项目:
上海市科委科研计划项目（15142200200，16142200300）；中国博士后科学基金（2015M581628）；上海市引进技术的吸收与创新专项（XC-ZXSJ-02-2016-11）.

摘要: 以脂多糖类似物（KLA）诱导的RAW264.7细胞为研究对象，采用代谢组学研究手段，研究水飞蓟素对脂多糖诱导炎症模型中花生四烯酸代谢通路的影响。以超高效液相色谱-三重四极杆质谱联用为平台，对不同浓度水飞蓟素作用下KLA诱导RAW264.7炎症细胞分泌的类二十烷酸代谢物进行定量分析，通过考察主成分分析（PCA）、正交偏最小二乘法判别分析（OPLS-DA）的VIP值和Kruskal-Wallis秩和检验结果显著性差异（P）值筛选代谢标记物。建立了59种类二十烷酸（含15种同位素内标）在5 min内实现快速分离的液相色谱-质谱联用方法；确定了细胞存活率在58%~80%的水飞蓟素浓度为50~150 μmol/L；筛选出数据处理结果同时满足变异权重参数（VIP）值> 1且结果P值< 0.05的类二十烷酸代谢标记物12-OxoLeukotriene B₄（12-OxoLTB₄）；通过分析柱状图和炎症信号通路，确定水飞蓟素借助其抗氧自由基特性发挥抗炎作用，通过抑制脂氧合酶-5（5-LOX）的活性及阻断5-LOX代谢通路中产生氧自由基的脂质过氧化反应来减少氧自由基及过氧化物的形成。综上所述，所建立的方法能快速准确地定量分析多种类二十烷酸，并从代谢组学角度解释了水飞蓟素的抗炎机制。

关键词:

English

Investigation on silymarin impact on lipopolysaccharide induced inflammation model based on arachidonic acid metabolism pathway

School of Pharmacy, Shanghai Jiao Tong University, Shanghai 200210, China
Received Date: 20 January 2017
Fund Project:
Shanghai Science and Technology Commission Scientific Research Project (Nos. 15142200200, 16142200300)

Abstract: The objective of this research is to investigate the suppressive effect of silymarin on vitro cell culture model of inflammatory macrophage RAW264.7 induced by Kdo2-Lipid A, and explore its mechanism based on cell metabonomics. Ultra-high performance liquid chromatography coupled with tandem mass spectrometry (UPLC-MS/MS) method was used in the cell metabonomic assay to quantitative analysis of metabolites related to eicosanoids pathway. Then chemometric approaches such as principal component analysis were used to process the metabolic data. Within the established method, a total of 59 eicosanoids standards (containing 15 deuterated internal standards) were simultaneously separated in a single 5 min run, and the analytical method is proved to be rapid, sensitive and accurate. Whereafter, the metabolites with VIP> 1 and P value< 0.05 were considered as biomarkers. 12-OxoLeukotriene B₄ (12-OxoLTB₄) was eventually identified as metabolic biomarkers of silymarin treatment group in this research, and according to the related inflammatory pathways, we speculated silymarin has anti-inflammatory activities by inhibiting the 5-lipoxygenase (5-LOX) activity and blocking lipid peroxidation in 5-LOX metabolic pathways to reduce the formation of peroxides and oxygen free radicals. This study provide a novel approach to the mechanism research on the silymarin treatment on RAW264.7 cells based on cell metabonomics.

Key words:

ultra-high performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS)
/ eicosanoids
/ deuterated internal standards
/ silymarin
/ metabonomics
/ inflammatory

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基于花生四烯酸代谢通路研究水飞蓟素对脂多糖诱导炎症模型影响

English

Investigation on silymarin impact on lipopolysaccharide induced inflammation model based on arachidonic acid metabolism pathway

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通讯作者: 陈斌, bchen63@163.com

中国化学会秘书处

基于花生四烯酸代谢通路研究水飞蓟素对脂多糖诱导炎症模型影响

English

Investigation on silymarin impact on lipopolysaccharide induced inflammation model based on arachidonic acid metabolism pathway

CCS Chemistry：嵌段共聚物自组装成 “三明治”二维有机材料，实现纳米级压力传感功能

CCS Chemistry：颜徐州、鲍哲南： 你的皮肤可能是一片SPMs

CCS Chemistry：工作高效不疲劳，只须让催化剂动起来

CCS Chemistry：静电组装导向聚合- 聚电解质纳米凝胶量化制备新策略

CCS Chemistry：金黄色葡萄球菌醛基脱氢酶是如何识别特异性底物的？

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通讯作者: 陈斌, bchen63@163.com

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