Generation of reactive oxygen species (ROS) is a key factor for stimulation of macrophage proliferation by ceramide 1-phosphate
Generation of reactive oxygen species (ROS) is a key factor for stimulation of macrophage proliferation by ceramide 1-phosphate
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DOI：
10.1016/j.yexcr.2011.11.013
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发表时间：
2012-02-15
影响因子：
3.7
通讯作者：
Gomez-Munoz, Antonio
中科院分区：
文献类型：
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作者：
Arana, Lide;Gangoiti, Patricia;Gomez-Munoz, Antonio
我们以前证明，神经酰胺1-磷酸（C1 P）是成纤维细胞和巨噬细胞的促有丝分裂。然而，这一作用所涉及的机制仅部分描述。在这里，我们证明，C1 P刺激活性氧（ROS）的形成在原代骨髓来源的巨噬细胞，和ROS所需的C1 P的促有丝分裂作用。ROS的产生依赖于C1 P对NADPH氧化酶的预先激活，这是通过测量p4 Ophox亚基的磷酸化和p47 phox从胞质溶胶到质膜的转运来确定的。此外，C1 P激活胞浆钙依赖性磷脂酶A（2）和蛋白激酶C-α，NADPH氧化酶的激活被这些酶的选择性抑制剂阻断。这些抑制剂和ROS产生的抑制剂阻断了C1 P的促有丝分裂作用。通过使用BHNB-C1 P（一种光不稳定的笼状C1 P类似物），我们证明了所有这些C1 P作用都是由细胞内C1 P引起的。可以得出结论，在骨髓源性巨噬细胞中负责C1 P刺激的ROS生成的酶是NADPH氧化酶，并且该酶在该途径中位于PKC-α和cPLA（2）-α的下游。(C)2011 Elsevier Inc. All rights reserved.
We previously demonstrated that ceramide 1-phosphate (C1P) is mitogenic for fibroblasts and macrophages. However, the mechanisms involved in this action were only partially described. Here, we demonstrate that C1P stimulates reactive oxygen species (ROS) formation in primary bone marrow-derived macrophages, and that ROS are required for the mitogenic effect of C1P. ROS production was dependent upon prior activation of NADPH oxidase by C1P, which was determined by measuring phosphorylation of the p4Ophox subunit and translocation of p47phox from the cytosol to the plasma membrane. In addition, C1P activated cytosolic calcium-dependent phospholipase A(2) and protein kinase C-alpha, and NADPH oxidase activation was blocked by selective inhibitors of these enzymes. These inhibitors, and inhibitors of ROS production, blocked the mitogenic effect of C1P. By using BHNB-C1P (a photolabile caged-C1P analog), we demonstrate that all of these C1P actions are caused by intracellular C1P. It can be concluded that the enzyme responsible for C1P-stimulated ROS generation in bone marrow-derived macrophages is NADPH oxidase, and that this enzyme is downstream of PKC-alpha and cPLA(2)-alpha in this pathway. (C) 2011 Elsevier Inc. All rights reserved.