Pannexin1 Channel Inhibitor ((10)panx) Protects Against Transient Focal Cerebral Ischemic Injury by Inhibiting RIP3 Expression and Inflammatory Response in Rats
Pannexin1 Channel Inhibitor ((10)panx) Protects Against Transient Focal Cerebral Ischemic Injury by Inhibiting RIP3 Expression and Inflammatory Response in Rats
复制标题
Pannexin1 通道抑制剂 ((10)panx) 通过抑制 RIP3 表达和炎症反应来防止大鼠短暂性局灶性脑缺血损伤
DOI：
10.1016/j.neuroscience.2020.02.042
复制
发表时间：
2020
期刊：
影响因子：
3.3
通讯作者：
Luo Benyan
中科院分区：
文献类型：
--
作者：
Wei Ruili;Bao Wangxiao;He Fangping;Meng Fangxia;Liang Hui;Luo Benyan
背景：近年来的研究表明程序性坏死（necroptosis）是缺血性神经元损伤的迟发性组成部分，而我们前期的研究表明pannexin 1通道参与脑缺血损伤和细胞炎症反应。在这里，我们研究了泛连接蛋白1通道抑制剂10 panx是否可以通过抑制细胞坏死和相关炎症来减轻大鼠局灶性缺血性脑损伤。雄性Sprague-Dawley大鼠随机分为假手术组、短暂性大脑中动脉闭塞（MCAO）组和10个潘生丁治疗组。我们通过评估梗死体积和神经功能缺损来研究10 panx的作用。此外，我们使用免疫荧光染色，蛋白质印迹，酶联免疫吸附试验（ELISA）和TUNEL检测来确定潜在的机制。结果表明，10 panx可减少脑梗死体积，减轻脑缺血后神经功能缺损，改善脑缺血后神经元死亡，但不能减少TUNEL阳性神经元和c-caspase 3的表达。相比之下，坏死性凋亡相关蛋白受体相互作用蛋白3（RIP 3）的表达显着降低。此外，10 panx减少神经元高迁移率族蛋白1（HMGB 1）的释放，抑制小胶质细胞活化和促炎因子的分泌。RIP 3与HMGB 1的免疫共标记结果表明，RIP 3蛋白与HMGB 1从细胞核向细胞质的释放密切相关。我们的数据表明，10 panx治疗可能通过减少RIP 3介导的坏死性凋亡，HMGB 1释放和相关的炎症反应来减轻MCAO损伤。RIP 3可能在脑卒中后HMGB 1的释放和炎症反应中起重要作用。
Background:Recent studies have demonstrated that programmed necrosis (necroptosis) is a delayed component of ischemic neuronal injury and our previous study has shown that pannexin 1 channel is involved in cerebral ischemic injury and cellular inflammatory response. Here, we examined whether the pannexin 1 channel inhibitor,10panx, could reduce focal ischemic brain injury in rats by inhibiting cellular necroptosis and the associated inflammation. Male Sprague–Dawley rats were randomly divided into sham-operated, MCAO (transient middle cerebral artery occlusion) group, and10panx-treated groups. We investigated the effect of10panx by assessing infarct volume and neurological deficit. Further, we determined the potential mechanism using immunofluorescent staining, Western blotting, enzyme-linked immunosorbent assay (ELISA) and TUNEL assay. We demonstrated that10panx reduced infarct volume and alleviated neurological deficit in the MCAO injury model.10panx ameliorated post-ischemic neuronal death, but it did not reduce the TUNEL positive neurons and expression of cleaved-caspase3. In contrast, expression of necroptosis related protein receptor-interacting protein 3 (RIP3) was significantly decreased. Furthermore,10panx reduced the release of high mobility group box 1 (HMGB1) from neurons and inhibited microglial activation and secretion of pro-inflammatory factors. Immunent co-labeling of RIP3 with HMGB1 showed that RIP3 protein was closely related with the release of HMGB1 from nucleus to cytoplasm. Our data suggested that10panx treatment may ameliorate MCAO injury by reducing RIP3-mediated necroptosis, HMGB1 release and associated inflammatory response. RIP3 may play an important role in the release of HMGB1 and inflammation after stroke.