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
发布时间：2019-01-21 18:49
【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50~150μmol/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。
[Abstract]:The effects of silymarin on arachidonic acid metabolic pathway in lipopolysaccharide induced inflammatory model were studied in RAW264.7 cells induced by lipopolysaccharide analogues (KLA). Using ultra-high performance liquid chromatography-quadrupole mass spectrometry as a platform, the eicosanoic acid metabolites secreted by RAW264.7 inflammatory cells induced by KLA with different concentrations of silymarin were quantitatively analyzed. The principal component analysis of (PCA), was carried out. The VIP value of orthogonal partial least squares discriminant analysis (OPLS-DA) was significantly different from that of Kruskal-Wallis rank sum test. (P) value was used to screen metabolic markers. A rapid separation method of 59 eicosanoic acids (including 15 isotopic internal standards) within 5 min was established by liquid chromatography-mass spectrometry, and the concentration of silymarin with cell survival rate of 58 ~ 80% was determined to be 50 ~ 150 渭 mol/L;. The eicosanoic acid metabolic marker 12-OxoLeukotriene B4 (12-OxoLTB4), which satisfied the variation weight parameter (VIP) value 1 and the result P value 0. 05, was screened out. By analyzing the histogram and the inflammatory signaling pathway, silymarin was determined to play an anti-inflammatory effect with the help of its anti-oxygen free radical properties. The formation of oxygen free radicals and peroxides was reduced by inhibiting the activity of lipoxygenase-5 (5-LOX) and blocking the lipid peroxidation of oxygen free radicals in the metabolic pathway of 5-LOX. In conclusion, the established method can be used for rapid and accurate quantitative analysis of many kinds of eicosanoic acids, and explains the anti-inflammatory mechanism of silymarin from the perspective of metabolomics.
【作者单位】： 上海交通大学药学院;
【基金】：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)~~
【分类号】：O657.63;R284
本文编号：2412931
[Abstract]:The effects of silymarin on arachidonic acid metabolic pathway in lipopolysaccharide induced inflammatory model were studied in RAW264.7 cells induced by lipopolysaccharide analogues (KLA). Using ultra-high performance liquid chromatography-quadrupole mass spectrometry as a platform, the eicosanoic acid metabolites secreted by RAW264.7 inflammatory cells induced by KLA with different concentrations of silymarin were quantitatively analyzed. The principal component analysis of (PCA), was carried out. The VIP value of orthogonal partial least squares discriminant analysis (OPLS-DA) was significantly different from that of Kruskal-Wallis rank sum test. (P) value was used to screen metabolic markers. A rapid separation method of 59 eicosanoic acids (including 15 isotopic internal standards) within 5 min was established by liquid chromatography-mass spectrometry, and the concentration of silymarin with cell survival rate of 58 ~ 80% was determined to be 50 ~ 150 渭 mol/L;. The eicosanoic acid metabolic marker 12-OxoLeukotriene B4 (12-OxoLTB4), which satisfied the variation weight parameter (VIP) value 1 and the result P value 0. 05, was screened out. By analyzing the histogram and the inflammatory signaling pathway, silymarin was determined to play an anti-inflammatory effect with the help of its anti-oxygen free radical properties. The formation of oxygen free radicals and peroxides was reduced by inhibiting the activity of lipoxygenase-5 (5-LOX) and blocking the lipid peroxidation of oxygen free radicals in the metabolic pathway of 5-LOX. In conclusion, the established method can be used for rapid and accurate quantitative analysis of many kinds of eicosanoic acids, and explains the anti-inflammatory mechanism of silymarin from the perspective of metabolomics.
【作者单位】： 上海交通大学药学院;
【基金】：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)~~
【分类号】：O657.63;R284
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1 王萍,丛润滋,王俊德,张乐沣;水飞蓟素中活性黄酮的测定[J];色谱;1998年06期
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