Glibenclamide-sulfonylurea receptor 1 antagonist alleviates LPS-induced BV2 cell activation through the p38/MAPK pathway
Glibenclamide-sulfonylurea receptor 1 antagonist alleviates LPS-induced BV2 cell activation through the p38/MAPK pathway
复制标题
格列本脲-磺酰脲受体 1 拮抗剂通过 p38/MAPK 途径减轻 LPS 诱导的 BV2 细胞活化
作者：
Xu Zhiming;Liu Yingliang;Yang Dianxu;Yuan Fang;Ding Jun;Tian Hengli;Wang Liping;Qu Meijie;Yang Guoyuan;Yang Guoyuan;Tian HL
我们研究了格列本脲，磺酰脲受体1（Sur 1）拮抗剂，在体外对LPS诱导的小胶质细胞活化的抗神经炎症活性和机制。将BV 2小胶质细胞暴露于LPS（100 ng mL-1）。采用RT-PCR和Western blotting检测iNOS和考克斯-2水平、促炎细胞因子mRNA表达以及p38/MAPK信号通路。格列本脲（2.5、10和40 μM）预处理可抑制LPS诱导的BV 2小胶质细胞中iNOS和考克斯-2的过度表达。阻断Sur 1可降低细胞内活性氧（ROS）水平。格列本脲剂量依赖性（2.5、10 μM）降低LPS诱导的TNF-α、IL-6和IL-1β的过度表达，并减轻LPS处理的BV 2小胶质细胞中的细胞内钙蓄积。此外，格列本脲还能抑制LPS诱导的p38/MAPK磷酸化，p38/MAPK选择性抑制剂SB 203580能显著增强格列本脲对LPS诱导的BV 2细胞iNOS和考克斯-2表达的抑制作用。格列本脲-Sur 1拮抗剂通过抑制p38/MAPK信号通路和促炎反应在体外小鼠小胶质细胞中发挥抗炎活性。格列本脲可能被开发为抑制中枢神经系统炎症反应的新型药物。
We investigated the anti-neuroinflammatory activity and mechanism of glibenclamide, sulfonylurea receptor 1 (Sur1) antagonist, against LPS-induced microglial activation in vitro. BV2 microglia cells were exposed to LPS (100 ng mL−1). iNOS and COX-2 levels, proinflammatory cytokine mRNA expression, and the p38/MAPK signaling pathway were analyzed by RT-PCR and Western blotting. Pretreatment with glibenclamide (2.5, 10, and 40 μM) inhibited the LPS-induced overexpression of iNOS and COX-2 in BV2 microglia cells. Blocking Sur1 reduced intracellular reactive oxygen species (ROS) levels. Glibenclamide dose-dependently (2.5, 10 μM) decreased LPS-induced over-expression of TNF-α, IL-6, and IL-1β, and alleviated the intracellular calcium accumulation in LPS-treated BV2 microglia cells. Moreover, glibenclamide diminished the LPS-induced phosphorylation of p38/MAPK, SB203580, a selective p38/MAPK inhibitor, significantly potentiated glibenclamide-caused inhibition of the expression of iNOS and COX-2 in LPS-exposed BV2 cells. Glibenclamide–Sur1 antagonist exerts anti-inflammatory activity in murine microglia in vitro by inhibiting the p38/MAPK signaling pathways and proinflammatory responses. Glibenclamide may be developed as a novel agent for suppressing inflammatory responses in the central nervous system.