INTERLEUKIN-1 ENHANCES THE ABILITY OF CULTURED HUMAN UMBILICAL VEIN ENDOTHELIAL-CELLS TO OXIDIZE LINOLEIC-ACID
INTERLEUKIN-1 ENHANCES THE ABILITY OF CULTURED HUMAN UMBILICAL VEIN ENDOTHELIAL-CELLS TO OXIDIZE LINOLEIC-ACID
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DOI：
10.1074/jbc.270.29.17279
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发表时间：
1995-07-21
影响因子：
4.8
通讯作者：
VILA, L
中科院分区：
文献类型：
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作者：
CAMACHO, M;GODESSART, N;VILA, L
用重组白细胞介素(IL)-1 β处理人脐静脉内皮细胞（HUVEC），观察其对外源性亚油酸的代谢。高效液相色谱、气相色谱-质谱分析和手性分析表明，HUVEC酶催化亚油酸主要转化为13-(S)羟基-9(Z)、11(E)-十八烯二烯酸（13- hode）和9-(R)羟基-10(E)、12(Z)-十八烯二烯酸，并可能异构化为全反式化合物。IL-1 β以时间和剂量依赖的方式增加了所有十八烷酸的形成，EC相似（50）（约1单位/ml）。IL-1 β刺激细胞亚油酸的表观K-m值分别为15.59 +/- 8.39和152.9 +/- 84 μ M (P < 0.05)，表明IL-1 β刺激细胞亚油酸具有更高的底物亲和力。未经处理和IL-1 β处理的细胞产生的羟基十八烷酸的S/R对映体比率与分离的环加氧酶（cox）相似，而分离的15-脂加氧酶产生的13-HODE具有严格的S构型。在对照组和IL-1 β处理的细胞中，几种COX抑制剂以剂量依赖的方式抑制了十八烷酸的形成，COX2选择性抑制剂对IL-1 β处理的细胞比对照组更有效。IL-1 β处理后，COX1和COX2蛋白水平分别增加不到2倍和8倍。COX抑制剂的特异性已被证实，因为它们不抑制人多形核白细胞形成13-HODE。综上所述，这些结果表明，COXs参与了HUVEC中亚油酸的氧化代谢，IL-1 β通过诱导以COX2为主的新酶的表达来促进亚油酸的氧化代谢。
Human umbilical vein endothelial cells (HUVEC) were treated with recombinant interleukin (IL)-1 beta, and the metabolism of exogenous linoleic acid was studied. High performance liquid chromatography, gas chromatography-mass spectrometry, and chiral analysis revealed that HUVEC enzymatically convert linoleic acid mainly into 13-(S)hydroxy-9(Z),11(E)-octadecadienoic (13-HODE) and 9-(R)hydroxy-10(E),12(Z)-octadecadienoic acids, which may isomerize toward all-trans compounds. IL-1 beta increased the formation of all octadecanoids in a time and dose-dependent manner with similar EC(50) (approximately 1 unit/ml). The apparent K-m values of linoleic acid were 15.59 +/- 8.39 and 152.9 +/- 84 mu M (P < 0.05) in IL-1 beta-treated cells and controls, respectively, indicating a higher substrate affinity in cells stimulated with IL-1 beta. Ratios of S/R enantiomers for the hydroxyoctadecanoids produced by untreated and IL-1 beta-treated cells were similar to those from isolated cyclooxygenases (COXs), whereas isolated 15-lipoxygenase yielded 13-HODE with a strict S configuration. The formation of octadecanoids was inhibited in a dose dependent manner by several COX inhibitors in both controls and IL-1 beta-treated cells, COX2 selective inhibitors being more effective on IL-1 beta-treated cells than on controls. COX1 and COX2 protein levels increased less than 2-fold and 8-fold, respectively, after IL-1 beta treatment. The specificity of COX inhibitors was proven since they did not inhibit 13-HODE formation by human polymorphonuclear leukocytes. Overall, these results indicate that COXs are responsible for the oxidative metabolism of linoleic acid in HUVEC, and IL-1 beta increases it by inducing the expression of new enzyme, mainly COX2.