Control of JNK for an activation of NADPH oxidase in LPS-stimulated BV2 microglia
Control of JNK for an activation of NADPH oxidase in LPS-stimulated BV2 microglia
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DOI：
10.1007/s12272-012-0415-1
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发表时间：
2012-03-01
影响因子：
6.7
通讯作者：
Choi, Ji Woong
中科院分区：
文献类型：
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作者：
Han, Jung Eun;Choi, Ji Woong
NADPH氧化酶是在各种CNS疾病下神经炎性细胞（包括小胶质细胞）中H2 O2产生的主要调节剂，并且其活性由丝裂原活化蛋白激酶（MAPK）控制，例如细胞外信号调节激酶1/2（ERK 1/2）、p38 MAPK和c-Jun N-末端激酶（JNK）。然而，很少有人知道NADPH氧化酶驱动的H2 O2生产力和JNK之间的联系在小胶质细胞。本研究的目的是使用脂多糖（LPS）刺激的BV 2小胶质细胞来揭示这种联系。LPS刺激的BV 2小胶质细胞产生的过氧化氢减少NADPH氧化酶抑制剂，包括4-（2-氨基乙基）苯磺酰氟和diphenyleniodonium氯化物。此外，NADPH氧化酶在LPS刺激的BV 2细胞中被激活。这些结果表明，在LPS刺激的BV 2小胶质细胞中，NAPDH氧化酶是H2 O2产生的主要因素。基于半定量PCR分析，两个NADPH氧化酶组分，p47（phox）和gp 91（phox），参与NADPH氧化酶的激活，因为这两个组分的转录水平被LPS上调。JNK在NADPH氧化酶调节的H2 O2生产中的作用是使用特异性抑制剂，包括SP 600125和JNK抑制肽（JIP）。抑制JNK通路显著降低H2 O2的产生，这与NADPH氧化酶活化的减弱和组分的上调密切相关。我们的结论是JNK途径参与NADPH氧化酶介导的过氧化氢生产力在BV 2小胶质细胞。
NADPH oxidase is a main regulator for H2O2 productivity in neuroinflammatory cells, including microglia, under various CNS diseases and its activity is controlled by mitogen-activated protein kinases (MAPKs), such as extracellular signal-regulated kinase 1/2 (ERK1/2), p38 MAPK, and c-Jun N-terminal kinase (JNK). However, little is known about the link between NADPH oxidase-driven H2O2 productivity and JNK in microglia. The purpose of this study is to uncover the link using lipopolysaccharide (LPS)-stimulated BV2 microglia. LPS-stimulated BV2 microglia produced H2O2 that was decreased by NADPH oxidase inhibitors, including 4-(2-aminoethyl)benzenesulfonylfluoride and diphenyleneiodonium chloride. In addition, NADPH oxidase was activated in LPS-stimulated BV2 cells. These results suggest that NAPDH oxidase is a main factor for H2O2 productivity in LPS-stimulated BV2 microglia. Based on a semi-quantitative PCR analysis, two of NADPH oxidase components, p47(phox) and gp91(phox), were involved in the activation of NADPH oxidase because transcriptional levels of both components were upregulated by LPS. Role of JNK in NADPH oxidase-regulated H2O2 productivity was pursued using specific inhibitors, including SP600125 and JNK inhibitory peptide (JIP). Inhibition of the JNK pathways significantly reduced H2O2 productivity, which was closely related to the attenuation of NADPH oxidase activation and the upregulation of components. We conclude that JNK pathways are involved in NADPH oxidase-mediated H2O2 productivity in BV2 microglia.