C-X3-C基序趋化因子配体1/C-X3-C基序趋化因子受体1通路参与失血性休克/复苏大鼠记忆功能恢复  

C-X3-C motif chemokine ligand 1/C-X3-C motif chemokine receptor 1 pathway in the recovery of memory function in hemorrhagic shock/resuscitation rats

作　　者：吴小军 王日兴[1] 林芳崇 吕有凯 冯奇桃 云天奇 WU Xiao-jun;WANG Ri-xing;LIN Fang-chong;LÜYou-kai;FENG Qi-tao;YUN Tian-qi(Department of Emergency,the Second Affiliated Hospital of Hainan Medical College,Haikou 570311,China)

机构地区：[1]海南医学院第二附属医院急诊科,海口570311

出　　处：《解剖学报》2024年第2期158-166,共9页Acta Anatomica Sinica

基　　金：海南省卫生健康行业科研项目(21A200312)。

摘　　要：目的探讨C-X3-C基序趋化因子配体1(CX3CL1)/C-X3-C基序趋化因子受体1(CX3CR1)通路调控的小胶质细胞活化对失血性休克/复苏大鼠记忆功能的影响。方法实验分为2部分。第1部分,将大鼠随机分为假手术组,模型-0.5 h组,模型-1.5 h组,模型-3 h组,每组10只,模型组之间失血性休克时间存在差异。第2部分,大鼠随机分为对照组与CX3CL1组,每组10只。CX3CL1组大鼠脑室注射CX3CL1蛋白,对照组大鼠注射生理盐水。所有大鼠在模型制作前开展Morris水迷宫训练,模型制作后4 d,开展水迷宫测试。完成后,取全脑组织进行HE染色与免疫组织化学染色,取脑脊液检测炎性细胞因子含量,取脑组织进行Real-time PCR检测与Western blotting检测。结果与假手术组相比,模型组大鼠逃避潜伏期增加,穿越平台次数与第Ⅲ象限停留时间减少,且HE染色中显示神经元状态受损。此外,与假手术组相比,模型组大鼠脑组织中离子钙结合衔接分子1(Iba1)表达升高,脑脊液中肿瘤细胞坏死因子α(TNF-α)与白细胞介素(IL)-6含量升高,M1型小胶质细胞标记CD16、TNF-α、IL-1β与诱导性一氧化氮合酶(iNOS)mRNA含量升高。与此同时,与假手术组相比,模型组大鼠脑组织中CX3CL1、CX3CR1蛋白表达降低,磷酸化核因子κB(p-NF-κB)与核苷酸结合寡聚结构域(NOD)样受体蛋白3(NLRP3)蛋白表达升高。然而,与模型组相比,CX3CL1组大鼠逃避潜伏期减少,穿越平台次数与第Ⅲ象限停留时间增加,且神经元状态恢复。此外,与对照组相比,CX3CL1组大鼠脑组织中Iba1表达降低,脑脊液中TNF-α与IL-6含量降低,M1型小胶质细胞标记CD16、TNF-α、IL-1β与iNOS mRNA含量降低,M2型小胶质细胞标记CD206、转化生长因子β(TGF-β)、精氨酸酶1(Arg1)、几丁质酶3样蛋白1(Ym1)mRNA含量升高。结论CX3CL1有助于抑制小胶质细胞过度激活,诱导小胶质细胞向M2型极化,抑制M1型极化,降低炎性细胞因子释放,减轻失�Objective To investigate the effect of microglia activation regulated by C-X3-C motif chemokine ligand 1(CX3CL1)-C-X3-C motif chemokine receptor 1(CX3CR1)pathway on memory function in hemorrhagic shock/resuscitation rats.Methods The experiment was divided into two parts.In the first part,the rats were randomly divided into sham group,model-0.5 hour group,model-1.5 hour group,model-3 hour group,10 rats in each group.There were differences in the time of hemorrhagic shock among each group.In the second part,rats were randomly divided into control group and CX3CL1 group,10 rats in each group.The rats in CX3CL1 group were treated with CX3CL1 protein factor(intraventricular injection),and the rats in control group were treated with saline.All rats were trained in Morris water maze experiments before model construction,and tests of Morris water maze experiments were carried out after 4 days of model construction.After completion,the whole brains were taken for HE staining and immunohistochemical staining.Cerebrospinal fluid was taken for detection of inflammatory cytokines,and hippocampus tissues were taken for Real-time PCR detection and Western blotting detection.Results Compared with the sham group,the escape latency of rats in model group increased,the number of platform crossings and the resident time in the third quadrant decreased.The neuronal state was impaired in HE staining in model group.In addition,compared with the sham group,the expression of ionized calcium binding adaptor molecule-1(Iba1)in the brain of the rats in model group increased,the contents of tumor necrosis factor-α(TNF-α)and interleukin(IL)-6 in the cerebrospinal fluid increased,and the M1-type microglia markers CD16,TNF-α,IL-1βand inducible nitric oxide synthase(iNOS)mRNA content increased.At the same time,compared with the sham group,the expressions of CX3CL1 and CX3CR1 in the brain of model group decreased,and the expressions of phosphorylated nuclear factor-κB(p-NF-κB)and nucleotide binding oligomerization domain(NOD)-like receptor

关 键 词：失血性休克/复苏 C-X3-C基序趋化因子配体1 记忆功能 炎症反应 小胶质细胞 实时定量聚合酶链反应 大鼠 

分 类 号：R605.97[医药卫生—急诊医学] R743.3[医药卫生—外科学]