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首页> 外文期刊>Clinical and diagnostic laboratory immunology >Role of Arachidonic Acid and Its Metabolites in the Priming of NADPH Oxidase in Human Polymorphonuclear Leukocytes by Peritoneal Dialysis Effluent
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Role of Arachidonic Acid and Its Metabolites in the Priming of NADPH Oxidase in Human Polymorphonuclear Leukocytes by Peritoneal Dialysis Effluent

机译：腹膜透析液中花生四烯酸及其代谢产物在人多形核白细胞NADPH氧化酶启动中的作用

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Peritoneal dialysis effluent (PDE) contains a low-molecular-weight solute that will activate and prime the NADPH oxidase of human neutrophils via a phospholipase A2(PLA2)-dependent mechanism. Since the products of PLA2 are known to activate and prime the oxidase we have investigated their role in the dialysis effluent-mediated activation and priming of human neutrophils. NADPH oxidase activity of PDE-primed and -unprimed neutrophils was measured by lucigenin-enhanced chemiluminescence in the presence of known inhibitors of the arachidonic acid cascade. Incubation of neutrophils with the nonselective PLA2 inhibitor quinacrine (0 to 100 μM) reduced oxidase activity in both primed and unprimed cells. Furthermore, primed cells were more sensitive to the action of quinacrine than were unprimed cells. We were unable to determine the relative roles of secretory PLA2(sPLA2) and cytosolic PLA2(cPLA2) since the selectivesPLA2 inhibitor scalaradial (0 to 100 μM) inhibited oxidase activity in both groups of cells by similar degrees, while the specific cPLA2 inhibitor AACO-CF3 (0 to 50 μM) failed to affect activity in either group. Inhibition of platelet-activating factor (PAF), cycloxygenase, and 5-lipoxygenase-activating protein by hexanolamino-PAF (0 to 25 μM), flurbiprofen (0 to 25 μM), and MK886 (0 to 5 μM), respectively, had no effect upon oxidase activity. However, the direct inhibition of 5-lipoxygenase by caffeic acid or lipoxin A4resulted in a similar concentration-dependent attenuation of oxidase activity in both primed and unprimed cells. Leukotriene B4(LTB4) release from primed neutrophils was comparable to that from unprimed cells with the exception of phorbol myristate acetate-stimulated cells, which released fivefold more LTB4than control. Taken together, these results suggest that it is arachidonic acid per se, and not its metabolites, that is important in priming of the neutrophil NADPH oxidase by dialysis effluent.
机译：腹膜透析流出物（PDE）包含一种低分子量溶质，可通过磷脂酶A 2 （PLA 2 ）依赖性激活并启动人中性粒细胞的NADPH氧化酶机制。由于已知PLA 2 的产物可以激活和引发氧化酶，因此我们研究了它们在透析废水介导的人类嗜中性粒细胞激活和引发中的作用。在已知的花生四烯酸级联抑制剂存在下，通过光泽精增强的化学发光测定了PDE引发和未引发中性粒细胞的NADPH氧化酶活性。中性粒细胞与非选择性PLA 2 抑制剂奎纳克林（0至100μM）一起孵育可降低引发和未引发细胞的氧化酶活性。此外，与未启动细胞相比，已启动细胞对奎纳克林的作用更敏感。我们无法确定分泌性PLA 2 （ s PLA 2 ）和胞质PLA 2 （ c PLA 2 ），因为选择性 s PLA 2 抑制剂Scalaradial（0至100μM）抑制了氧化酶活性。两组细胞的程度相似，而特定的 c PLA 2 抑制剂AACO-CF 3 （0至50μM）不能影响活性在任何一组中。己糖胺氨基-PAF（0至25μM），氟比洛芬（0至25μM）和MK886（0至5μM）分别抑制了血小板激活因子（PAF），环氧化酶和5-脂氧化酶激活蛋白。对氧化酶活性没有影响。然而，咖啡因酸或脂蛋白A 4 对5-脂氧合酶的直接抑制作用导致了初免和初免细胞中氧化酶活性的类似浓度依赖性减弱。引发的中性粒细胞释放的白三烯B 4 （LTB 4 ）与未引发的细胞相当，但佛波醇肉豆蔻酸酯乙酸盐刺激的细胞除外，后者释放的LTB 4 > 4 胜过控制。总之，这些结果表明，花生四烯酸本身而不是其代谢产物在通过透析流出液引发中性粒细胞NADPH氧化酶中起着重要的作用。

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