TLR4-mediated MyD88-dependent signaling pathway is activated by cerebral ischemia-reperfusion in cortex in mice
TLR4-mediated MyD88-dependent signaling pathway is activated by cerebral ischemia-reperfusion in cortex in mice
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DOI：
10.1016/j.biopha.2008.06.028
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发表时间：
2009-07-01
影响因子：
7.5
通讯作者：
Zhang, Baohui
中科院分区：
文献类型：
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作者：
Gao, Yin;Fang, Xiubin;Zhang, Baohui
探讨脑缺血再灌注炎症反应中信号通路是否被激活及其机制。将小鼠随机分为假手术组、缺血再灌注组和TLR4阻断组，再灌注不同时间点为12 h、24 h、48 h和72 h组。观察缺血再灌注后不同时间点各组TLR4 mRNA和MyD88 mRNA表达、NF-κB活化以及TNF-α和IL-1β蛋白水平的差异。通过双侧颈总动脉（CCA）闭塞诱导小鼠脑缺血。 TLR4信号通路可以被特异性抗TLR4结合蛋白抑制，从而阻止TLR4与其受体相互作用。我们通过Western blot检测TLR4抗体阻断和小鼠脑缺血再灌注损伤的结果，并评估皮质神经元损伤。我们还通过原位杂交（ISH）测定了TLR4 mRNA和MyD88 mRNA的表达，通过EMSA测定了NF-κB的激活，并通过Western blot测定了TNF-α蛋白的表达。再灌注前抗TLR4结合TLR4受体有效；各组间神经元损伤程度存在明显差异； TLR4 mRNA和MyD88 mRNA的表达、NF-κB的激活以及TNF-α蛋白的表达也显示出明显的差异。缺血再灌注激活的 LR4 介导的 MyD88 依赖性信号通路可能通过上调 NF-κ B 和 TNF-α 参与缺血再灌注机制。 (C) 2008 年由 Elsevier Masson SAS 出版。
To study whether the signaling pathway is activated in the inflammatory reaction of cerebral ischemia-reperfusion and its mechanism. The mice were randomly divided into sham group, ischemia-reperfusion group and TLR4-blocked group with different time points of reperfusion 12 h, 24 h, 48 h and 72 h group. We observed the different expression of TLR4 mRNA and MyD88 mRNA, activation of NF-kappa B and the TNF-alpha and IL-1 beta protein levels in each group at different time point after ischemia-reperfusion. Mice cerebral ischemia was induced by occlusion of common carotid arteries (CCA) bilaterally. TLR4 signaling pathway could be inhibited by specific anti-TLR4 binding protein to prevent TLR4 from interacting with its receptors. We determined the result of TLR4 antibodies-blocking and mice cerebral ischemia-reperfusion injuries by Western blot, and evaluated neuronal damage in cortex. We also determined the expression of TLR4 mRNA and MyD88 mRNA by in situ hybridization (ISH), the activation of NF-kappa B by EMSA, and the expression of TNF-alpha protein by Western blot. Anti-TLR4 binding TLR4 receptors before reperfusion was effective; There was distinct difference among each group respecting neuronal damage; The expression of TLR4 mRNA and MyD88 mRNA, the activation of NF-kappa B, and the expression of TNF-alpha protein showed clear difference as well. LR4-mediated MyD88-dependent signaling pathway activated by ischemia-reperfusion may be involved in the mechanism of ischemia-reperfusion through upregulation of NF-kappa B and TNF-alpha. (C) 2008 Published by Elsevier Masson SAS.