Acid‐sensing ion channels promote the inflammation and migration of cultured rat microglia
Acid‐sensing ion channels promote the inflammation and migration of cultured rat microglia
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DOI：
10.1002/glia.22766
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发表时间：
2015-03
期刊：
影响因子：
6.2
通讯作者：
Xiao-Wei Yu;Zhuang‐li Hu;Ming Ni;Peng Fang;Pei Zhang;Qing Shu;Hua Fan;Hai-Yun Zhou;Lan Ni;Ling-Qiang Zhu;Jian-Guo Chen;F. Wang
中科院分区：
文献类型：
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作者：
Xiao-Wei Yu;Zhuang‐li Hu;Ming Ni;Peng Fang;Pei Zhang;Qing Shu;Hua Fan;Hai-Yun Zhou;Lan Ni;Ling-Qiang Zhu;Jian-Guo Chen;F. Wang
小胶质细胞是中枢神经系统的主要免疫细胞，在免疫防御和炎症反应中起着监视和清道夫的作用。以往的研究表明，酸敏感离子通道（ASICs）与炎症可能有密切的关系，但ASICs在炎症过程中在小胶质细胞中的确切作用仍然是难以捉摸的。在本研究中，我们鉴定了原代培养的大鼠小胶质细胞中ASICs的存在，并探讨了它们的功能。通过逆转录聚合酶链反应（RT-PCR）、定量真实的-时间PCR（qPCR）、免疫印迹和免疫荧光实验，我们证明了在培养和原位的大鼠小胶质细胞中存在ASIC 1、ASIC 2a和ASIC 3。脂多糖（LPS）刺激后，小胶质细胞ASIC 1和ASIC 2a表达上调。同时，ASIC样电流和酸诱导的细胞内钙升高增加，这可以被非特异性ASIC拮抗剂阿米洛利和特异性同源ASIC 1a阻断剂PcTx 1抑制。此外，这两种抑制剂减少炎症细胞因子的表达，包括诱导型一氧化氮合酶和环氧合酶2由LPS刺激。此外，我们还观察到在刮擦刺激的小胶质细胞迁移中ASIC 1和ASIC 2a的表达显著增加。阿米洛利和PcTx 1通过抑制ERK磷酸化而阻止迁移。综上所述，这些结果表明ASIC参与神经炎症反应，这将为控制炎症相关神经元疾病提供新的治疗策略。GLIA 2015;63：483-496
Microglia, the major immune cells in central nervous system, act as the surveillance and scavenger of immune defense and inflammatory response. Previous studies suggest that there might be close relationship between acid‐sensing ion channels (ASICs) and inflammation, however, the exact role of ASICs in microglia during inflammation remains elusive. In the present study, we identified the existence of ASICs in the primary cultured rat microglia and explored their functions. By using reverse transcriptase polymerase chain reaction (RT‐PCR), quantitative real‐time PCR (qPCR), western blotting, and immunofluorescence experiments, we demonstrated that ASIC1, ASIC2a, and ASIC3 were existed in cultured and in situ rat microglia. After lipopolysaccharide (LPS) stimulation, the expressions of microglial ASIC1 and ASIC2a were upregulated. Meanwhile, ASIC‐like currents and acid‐induced elevation of intracellular calcium were increased, which could be inhibited by the nonspecific ASICs antagonist amiloride and specific homomeric ASIC1a blocker PcTx1. In addition, both inhibitors reduced the expression of inflammatory cytokines, including inducible nitric oxide synthase and cyclooxygenase 2 stimulated by LPS. Furthermore, we also observed significant increase in the expression of ASIC1 and ASIC2a in scrape‐stimulated microglial migration. Amiloride and PcTx1 prevented the migration by inhibiting ERK phosphorylation. Taken together, these results suggest that ASICs participate in neuroinflammatory response, which will provide a novel therapeutic strategy for controlling the inflammation‐relevant neuronal diseases. GLIA 2015;63:483–496