收稿日期: 2024-02-21
网络出版日期: 2024-07-15
基金资助
湖北省卫生健康科研基金项目(H20210036)
The neuroprotective mechanism of RKIP signaling pathway expression on microglial polarization in cerebral hemorrhage rat
Received date: 2024-02-21
Online published: 2024-07-15
目的 探讨Raf激酶抑制剂蛋白（RKIP）介导的小胶质细胞极化在脑出血（ICH）模型中的神经保护作用。 方法 48只成年雄性Sprague-Dawley （SD）大鼠随机分成3组：Sham + Vector组、ICH + Vector组和ICH + RKIP组，每组16只。ICH + Vector组和ICH + RKIP组建立胶原酶ICH模型。在手术前及手术后1、3、5和7 d，每组有8只动物行为测试。通过流式细胞术检测神经元凋亡情况。在ICH后7 d，通过蛋白质印迹分析血肿周围RKIP、p-p65、TRAF6表达。 结果 与ICH + Vector组相比，ICH + RKIP组大鼠找到平台的时间显著缩短，并且在目标象限中花费时间和跨平台次数显著增加（P < 0.05）。ICH + RKIP组Nissl小体的数量显著高于ICH + Vector组（P < 0.05）。此外，ICH + RKIP组神经元凋亡数量显著低于ICH+Vector组（P < 0.05）。与Sham组相比，接受ICH的大鼠表现出RKIP表达逐渐降低，并在第7天达到最低值（P < 0.05）。在ICH后7 d，ICH + RKIP组大鼠血肿中RKIP蛋白表达较ICH + Vector组显著增加（P < 0.05），p-p65、TRAF6蛋白表达较ICH + Vector组显著降低（P < 0.05）。与ICH + Vector组相比，ICH + RKIP组iNOS + Ibal1+细胞数目显著降低（P < 0.05），和Arg-1 + Ibal1+细胞数目显著增加（P < 0.05）。 结论 RKIP上调促进ICH后的功能恢复，其作用机制涉及抑制TRAF6/NF-κB信号通路。
关键词： Raf激酶抑制剂蛋白; 小胶质细胞; 极化; 脑出血
孙茹雪 , 朱梦莉 , 刘晶晶 , 陈飞 . Raf激酶抑制剂蛋白信号通路表达影响小胶质细胞极化对脑出血大鼠的神经保护机制[J]. 实用医学杂志, 2024 , 40(14) : 1935 -1940 . DOI: 10.3969/j.issn.1006-5725.2024.14.006
Objective This study aimed to investigate the neuroprotective effect of microglia polarization mediated by Raf kinase inhibitor protein （RKIP） intracerebral hemorrhage （ICH） model. Methods Forty-eight adult male Sprague-Dawley （SD） rats were randomly divided into three groups：the Sham + Vector group，the ICH + Vector group， and the ICH + RKIP group， with 16 rats in each group. The collagenase ICH model was established in ICH+Vector group and ICH + RKIP group. Before operation and 1， 3， 5，and 7 days after operation， 8 animals in each group were tested for behavior. Apoptosis of neurons was detected by flow cytometry. Seven days after ICH， the expressions of RKIP， p-p65， and TRAF6 around hematoma were analyzed by protein blot. Results Compared with ICH + Vector group， rats in ICH + RKIP group need less time to find the platform， spend longer time in the target quadrant， and significantly reduce the times of crossing the platform （P < 0.05）. The number of Nissl corpuscles in ICH + RKIP group was significantly higher than that in ICH + Vector group （P < 0.05）. In addition， the number of neuronal apoptosis in ICH + RKIP group was significantly lower than that in ICH + Vector group （P < 0.05）. Compared with Sham group， rats receiving ICH showed a gradual decrease in RKIP expression， and reached the lowest value on the 7th day （P < 0.05）. Seven days after ICH， the expression of RKIP protein in hematoma of rats in ICH + RKIP group was significantly higher than that in ICH + Vector group （P < 0.05）， and the expression of p-p65 and TRAF6 protein was significantly lower than that in ICH + Vector group （P < 0.05）. Compared with ICH + Vector group， the number of iNOS + Ibal1+ cells in ICH + RKIP group decreased significantly （P < 0.05）， while the number of Arg-1 + Ibal1+ cells increased significantly （P < 0.05）. Conclusion Up-regulation of RKIP promotes functional recovery after ICH， and its mechanism involves inhibiting TRAF6/NF-κB signaling pathway.
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