n-6 and n-3 polyunsaturated fatty acids stimulate translocation of protein kinase C alpha, -beta I, -beta II and -epsilon and enhance agonist-induced NADPH oxidase in macrophages
n-6 and n-3 polyunsaturated fatty acids stimulate translocation of protein kinase C alpha, -beta I, -beta II and -epsilon and enhance agonist-induced NADPH oxidase in macrophages
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DOI：
10.1042/bj3250553
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发表时间：
1997-07-15
影响因子：
4.1
通讯作者：
Ferrante, A
中科院分区：
文献类型：
--
作者：
Huang, ZH;Hill, CST;Ferrante, A
多不饱和脂肪酸 (PUFA)、花生四烯酸 (AA)、二十碳五烯酸 (EPA) 和二十二碳六烯酸 (DHA) 是人类单核细胞和巨噬细胞中氧依赖性呼吸活动（化学发光）的不良诱导剂，但显着增强了对三肽 N-甲酰基甲硫氨酰-亮氨酰-苯丙氨酸的反应。这些脂肪酸的作用在浓度为 1 μg/ml 时可见。 PMA 也有类似的增强作用，PMA 是一种作用于蛋白激酶 C (PKC) 或钙离子载体 (A23187) 的刺激物，可增加细胞内的钙，表明脂肪酸的作用是表面受体结合后，在维生素 D-3 存在下通过培养分化为巨噬细胞样细胞的 HL-60 细胞也同样受到脂肪酸的影响。在改变单核细胞预暴露于PUFA的时间的实验中，发现PLA、EPA和DHA诱导的启动效应在5分钟时最大。如果脂肪酸被甲基化或氧化成氢和氢过氧衍生物，则这些脂肪酸与其他激动剂协同作用的能力将完全丧失。饱和脂肪酸无活性，蛋白质印迹分析表明，PUFA 诱导 PKC α、-β I、-β II 和 -ε 同工酶易位至颗粒部分。通过用 PKC 抑制剂 GF-109203X 预处理细胞，或用 PMA 预处理单核细胞 18 小时以降低 PKC 水平，可以完全抑制脂肪酸和 A23187 之间的协同反应。从这些研究中可以明显看出，PUFA 启动巨噬细胞，导致对其他刺激的氧化呼吸爆发活性增加/协同，并且这种启动依赖于 PKC 易位和激活。
The polyunsaturated fatty acids (PUFA), arachidonic acid (AA), eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA) were poor inducers of oxygen-dependent respiratory activity (chemiluminescence) in human monocytes and macrophages, but markedly enhanced the response to the tripeptide, N-formylmethionyl-leucyl-phenylalanine. The effects of these fatty acids were seen at concentrations of 1 mu g/ml. A similar enhancement was seen with PMA, a stimulus that acts on protein kinase C (PKC), or calcium ionophore (A23187), which increases intracellular calcium, suggesting that the effect of the fatty acids was post-surface receptor binding, HL-60 cells, differentiated to macrophage-like cells by culture in the presence of vitamin D-3, were similarly affected by the fatty acids. In experiments in which the time of pre-exposure of the monocytes to PUFA was varied, it was found that the priming effect induced by PLA, EPA and DHA was maximal at 5 min. The ability of these fatty acids to synergize with other agonists was completely lost if the fatty acids were either methylated or oxidized to the hydro and hydroperoxy derivatives. Saturated fatty acids were inactive, Western blot analysis demonstrated that the PUFA induced the translocation of PKC alpha, -beta I, -beta II and -epsilon isoenzymes to a particulate fraction. The synergistic response between fatty acids and A23187 was completely inhibited by pretreating the cells with a PKC inhibitor, GF-109203X, or by pretreatment of monocytes with PMA for 18 h, to deplete PKC levels. From these investigations it is evident that PUFA prime macrophages, causing increased/synergistic oxidative respiratory burst activity to other stimuli and that this priming is dependent on PKC translocation and activation.