Toll-Like Receptor 4-Mediated Myeloid Differentiation Factor 88-Dependent Signaling Pathway Is Activated by Cerebral Ischemia-Reperfusion in Hippocampal CA1 Region in Mice

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  • Gao Yin
    Department of Neurobiology, China Medical University Basic Medicine School, Qiqihar Medical University
  • Fang Xiubin
    Department of Neurobiology, China Medical University
  • Sun He
    Basic Medicine School, Qiqihar Medical University
  • Wang Yu
    Basic Medicine School, Qiqihar Medical University
  • Yao Li Jie
    Basic Medicine School, Qiqihar Medical University
  • Li Jing Ping
    Basic Medicine School, Qiqihar Medical University
  • Tong Yue
    Department of Neurobiology, China Medical University
  • Zhang Baohui
    Department of Neurobiology, China Medical University
  • Liu Yuli
    Department of Neurobiology, China Medical University

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The Toll-like receptor 4 (TLR4)-mediated myeloid differentiation factor 88 (MyD88)-dependent signaling pathway plays an essential role in inflammation resulting from invading microbes. However, whether the signaling pathway is activated in the inflammatory reaction of cerebral ischemia-reperfusion and its mechanism is still unclear. In this experiment mice were randomly divided into sham group, ischemia/reperfusion group and TLR4-blocked group with different time points of reperfusion at 12, 24, 48 and 72 h . Mice cerebral ischemia was induced by occlusion of common carotid arteries (CCA) bilaterally. TLR4 signaling pathway was inhibited using 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 the hippocampus. We also determined expression of TLR4 mRNA and MyD88 mRNA by in situ hybridization (ISH), activation of nuclear factor (NF)-κB by electrophoretic mobility-shift analysis (EMSA), and expression of interrleukin (IL)-1β protein by Western blot. The results demonstrated that TLR4-mediated MyD88-dependent signaling pathway activated by ischemia-reperfusion may be involved in the mechanism of ischemia-reperfusion through upregulation of NF-κB, IL-1β.

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