Molecular Mechanisms of N-Formyl-Methionyl-Leucyl-Phenylalanine-Induced Superoxide Generation and Degranulation in Mouse Neutrophils: Phospholipase D Is Dispensable
Molecular Mechanisms of N-Formyl-Methionyl-Leucyl-Phenylalanine-Induced Superoxide Generation and Degranulation in Mouse Neutrophils: Phospholipase D Is Dispensable
复制标题
DOI：
10.1128/mcb.00869-12
复制
发表时间：
2013-01-01
影响因子：
5.3
通讯作者：
Kanaho, Yasunori
中科院分区：
文献类型：
--
作者：
Sato, Takanobu;Hongu, Tsunaki;Kanaho, Yasunori
磷脂酶D（PLD），产生脂质信使磷脂酸（PA），已涉及超氧化物的产生和中性粒细胞的脱粒。这一结论的基础是观察到的伯醇，干扰PLD催化的PA生产，抑制这些中性粒细胞的功能。然而，不能完全排除伯醇的脱靶效应。在这里，我们产生PLD-/-小鼠，以重新评估PLD的参与，并研究这些中性粒细胞功能的分子机制。令人惊讶的是，N-甲酰-甲硫氨酰-亮氨酰-苯丙氨酸（fMLP）诱导PLD-/-中性粒细胞的这些功能，这些功能被抑制乙醇。这些结果表明，PLD是这些中性粒细胞的功能，乙醇非特异性抑制它们，警告使用伯醇作为PLD介导的PA形成的特异性抑制剂。钙离子载体离子霉素和膜渗透化合物1-油酰基-2-乙酰基-sn-甘油（OADG）协同诱导超氧化物的产生。另一方面，单独的离子霉素诱导脱粒，OADG进一步增强。这些结果表明，传统的蛋白激酶C（cPKC）是至关重要的超氧化物的产生，和钙依赖性信号通路（S）和cPKC参与在小鼠中性粒细胞的脱粒。
Phospholipase D (PLD), which produces the lipid messenger phosphatidic acid (PA), has been implicated in superoxide generation and degranulation in neutrophils. The basis for this conclusion is the observation that primary alcohols, which interfere with PLD-catalyzed PA production, inhibit these neutrophil functions. However, off-target effects of primary alcohols cannot be totally excluded. Here, we generated PLD-/- mice in order to reevaluate the involvement of PLD in and investigate the molecular mechanisms of these neutrophil functions. Surprisingly, N-formyl-methionyl-leucyl-phenylalanine (fMLP) induced these functions in PLD-/- neutrophils, and these functions were suppressed by ethanol. These results indicate that PLD is dispensable for these neutrophil functions and that ethanol nonspecifically inhibits them, warning against the use of primary alcohols as specific inhibitors of PLD-mediated PA formation. The calcium ionophore ionomycin and the membrane-permeative compound 1-oleoyl-2-acetyl-sn-glycerol (OADG) synergistically induced superoxide generation. On the other hand, ionomycin alone induced degranulation, which was further augmented by OADG. These results demonstrate that conventional protein kinase C (cPKC) is crucial for superoxide generation, and a Ca2+-dependent signaling pathway(s) and cPKC are involved in degranulation in mouse neutrophils.