CD200Fc reduces TLR4-mediated inflammatory responses in LPS-induced rat primary microglial cells via inhibition of the NF-κB pathway
CD200Fc reduces TLR4-mediated inflammatory responses in LPS-induced rat primary microglial cells via inhibition of the NF-κB pathway
复制标题
CD200Fc 通过抑制 NF-κB 通路减少 LPS 诱导的大鼠原代小胶质细胞中 TLR4 介导的炎症反应
DOI：
10.1007/s00011-016-0932-3
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发表时间：
2016-07-01
影响因子：
6.7
通讯作者：
Li, Min
中科院分区：
文献类型：
--
作者：
Jiang, Li;Xu, Fan;Li, Min
目的根据最近的资料，CD200R1激动剂CD200Fc可以减轻自身免疫性疾病和神经元变性过程中小胶质细胞的炎症反应。然而，CD200Fc在小胶质细胞中抗炎活性的确切分子机制尚未阐明。本研究探讨了CD200Fc对内毒素刺激的大鼠原代小胶质细胞的抗炎作用及其分子机制。结果CD200Fc和(或)β对小胶质细胞无明显的细胞毒性作用。用实时荧光定量聚合酶链式反应、免疫印迹和/或免疫荧光染色检测了NF-αB相关信号(MyD88、p-κ、TRIF、p-Tbk1、p-IRF3、p-IκB和NF-κB-p65)。脂多糖降低小胶质细胞CD200R1的表达，这种作用可被CD200Fc所减弱。此外，CD200Fc可抑制脂多糖诱导的小胶质细胞TLR4及其接头分子(MyD88和p-TAK1、TRIF、p-TBK1和p-IRF3)的表达，并阻断其与MyD88、TAK1和TRIF的相互作用。CD200Fc还可减轻脂多糖诱导的小胶质细胞p-IκB和NF-κB-p65蛋白的核转位。CD200Fc抑制脂多糖诱导的小胶质细胞炎性介质的释放，包括IL-1β、IL-6、肿瘤坏死因子-α、诱导型一氧化氮合酶、单核细胞趋化蛋白-1和环氧合酶-2。结论CD200Fc可能通过阻断TLR4介导的NF-κB活化而发挥抗炎作用。
ObjectiveBased on recent information, CD200Fc, a CD200R1 agonist, could attenuate the inflammatory response of microglial cells in autoimmune diseases and neuro-degeneration. However, the exact molecular mechanisms responsible for the anti-inflammatory activity of CD200Fc in microglial cells have not been elucidated. In the present study, we investigated the anti-inflammatory effects and the molecular mechanisms of CD200Fc in lipopolysaccharide (LPS)-stimulated rat primary microglial cells.MethodsThe cell viability was measured by MTT assay. The LPS-induced cytokines release (IL-1β, IL-6, TNF-α, iNOS, MCP-1, and COX-2) was monitored by ELISA or real-time PCR, while NF-κB-related signals (MyD88, p-TAK1, TRIF, p-TBK1, p-IRF3, p-IκB, and NF-κB-P65) were assessed by real-time PCR, western blot and/or Immunofluorescent staining.ResultsCD200Fc and/or LPS exerted no significant cytotoxicity on microglial cells. LPS reduced the CD200R1 expression in microglial cells, and this effect was attenuated by CD200Fc. In addition, CD200Fc inhibited LPS-induced expression of TLR4 and its adapter molecules (MyD88 and p-TAK1, TRIF, p-TBK1, and p-IRF3), and abolished its interactions with MyD88, TAK1, and TRIF in microglial cells. CD200Fc also attenuated LPS-induced protein expression of p-IκB and NF-κB-P65 translocation to nucleus in microglial cells. Moreover, CD200Fc suppressed the LPS-induced release of inflammatory mediators in microglial cells, including IL-1β, IL-6, TNF-α, iNOS, MCP-1, and COX-2.ConclusionThese results indicated that CD200Fc displayed an anti-inflammatory effect in LPS-induced microglial cells by blocking TLR4-mediated NF-κB activation.