Roles of Pannexin-1 Channels in Inflammatory Response through the TLRs/NF-Kappa B Signaling Pathway Following Experimental Subarachnoid Hemorrhage in Rats.
Roles of Pannexin-1 Channels in Inflammatory Response through the TLRs/NF-Kappa B Signaling Pathway Following Experimental Subarachnoid Hemorrhage in Rats.
复制标题
Pannexin-1 通道在大鼠实验性蛛网膜下腔出血后通过 TLRs/NF-Kappa B 信号通路的炎症反应中的作用。
DOI：
10.3389/fnmol.2017.00175
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发表时间：
2017
影响因子：
4.8
通讯作者：
Shi JX
中科院分区：
文献类型：
--
作者：
Wu LY;Ye ZN;Zhou CH;Wang CX;Xie GB;Zhang XS;Gao YY;Zhang ZH;Zhou ML;Zhuang Z;Liu JP;Hang CH;Shi JX
背景：越来越多的证据表明，神经炎症在蛛网膜下腔出血(SAH)后早期脑损伤中起重要作用。PAnnexin-1通道是位于质膜上的缝隙连接蛋白的一员，激活后释放ATP、离子、第二信使、神经递质和高达1kD的分子进入细胞外间隙。以往的研究表明，PAnnexin-1通道的开放对细胞迁移、细胞凋亡，尤其是炎症反应是必不可少的，但在SAH模型中，其对炎症反应的影响尚不清楚。方法：成年雄性SD大鼠分为假手术组(n=20)、SAH组(n=20)、SAH+LV-shRNA组(n=20)、SAH+LV-shRNA-Panx1组(n=20)、SAH+LV-NC组(n=20)和SAH+LV-Panx1-EGFP组(n=20)。将0.3ml新鲜动脉非肝素化血液注入20只S大鼠的交叉前池建立蛛网膜下腔出血模型，SAH+LV-shRNA-Panx1组和SAH+LV-Panx1-EGFP组经侧脑室注射慢病毒。在蛛网膜下腔出血诱导前72小时。采用实时定量聚合酶链式反应、凝胶迁移率改变分析、酶联免疫吸附分析、免疫荧光染色和Western blotting等方法，探讨Pannin-1通道与TLR2/TLR4/NF-κB介导的信号转导途径之间的相互作用机制。采用Morris水迷宫实验观察大鼠认知和记忆的变化。结果：LV-shRNA-Panx1能显著降低大鼠大脑皮层TLR2/4/NF-κB通路相关物质的表达水平，并能明显改善SAH模型大鼠的神经功能障碍。相反，给予LV-Panx1-EGFP后，TLR2/4/NF-κB通路相关因子的表达增加，这与神经元的凋亡增加有关。结论：PAnnexin-1通道可能通过TLR2/TLR4/NF-κB介导的信号通路参与蛛网膜下腔出血后的炎症反应和神经行为功能障碍，提示PAnnexin-1通道可能是治疗蛛网膜下腔出血的潜在靶点。
Background: Accumulating evidence suggests that neuroinflammation plays a critical role in early brain injury after subarachnoid hemorrhage (SAH). Pannexin-1 channels, as a member of gap junction proteins located on the plasma membrane, releases ATP, ions, second messengers, neurotransmitters, and molecules up to 1 kD into the extracellular space, when activated. Previous studies identified that the opening of Pannexin-1 channels is essential for cellular migration, apoptosis and especially inflammation, but its effects on inflammatory response in SAH model have not been explored yet. Methods: Adult male Sprague-Dawley rats were divided into six groups: sham group (n = 20), SAH group (n = 20), SAH + LV-Scramble-ShRNA group (n = 20), SAH + LV-ShRNA-Panx1 group (n = 20), SAH + LV-NC group (n = 20), and SAH + LV-Panx1-EGFP group (n = 20). The rat SAH model was induced by injection of 0.3 ml fresh arterial, non-heparinized blood into the prechiasmatic cistern in 20 s. In SAH + LV-ShRNA-Panx1 group and SAH + LV-Panx1-EGFP group, lentivirus was administered via intracerebroventricular injection (i.c.v.) at 72 h before the induction of SAH. The Quantitative real-time polymerase chain reaction, electrophoretic mobility shift assay, enzyme-linked immunosorbent assay, immunofluorescence staining, and western blotting were performed to explore the potential interactive mechanism between Pannexin-1 channels and TLR2/TLR4/NF-κB-mediated signaling pathway. Cognitive and memory changes were investigated by the Morris water maze test. Results: Administration with LV-ShRNA-Panx1 markedly decreased the expression levels of TLR2/4/NF-κB pathway-related agents in the brain cortex and significantly ameliorated neurological cognitive and memory deficits in this SAH model. On the contrary, administration of LV-Panx1-EGFP elevated the expressions of TLR2/4/NF-κB pathway-related agents, which correlated with augmented neuronal apoptosis. Conclusion: Pannexin-1 channels may contribute to inflammatory response and neurobehavioral dysfunction through the TLR2/TLR4/NF-κB-mediated pathway signaling after SAH, suggesting a potential role of Pannexin-1 channels could be a potential therapeutic target for the treatment of SAH.