脑出血与TLR4/NF-κB介导小胶质细胞自噬的相关性研究  

Correlationstudy between cerebral hemorrhage and TLR4/NF-κB mediated microgliaautophagy

作　　者：陆梦茹 朱祖福[1] 张慧萍[1] 孔玉[1] 高志强[1] 杨江胜[1] 陆强彬[1] 柏燕燕[1] 周国庆[1] 沈丽萍[1] Lu Meng-ru;Zhu Zufu;Zhang Huiping(Department of Neurology,Jiangyin People＇s Hospital,Jiangyin 214400)

机构地区：[1]江苏省江阴市人民医院神经内科,214400

出　　处：《卒中与神经疾病》2018年第4期373-376,共4页Stroke and Nervous Diseases

基　　金：2016年江阴科技局指令性计划项目澄政科(2016)37号(JYKJ3033)

摘　　要：目的探讨TLR4介导小胶质细胞自噬在脑出血后炎症反应中的作用及机制。方法 A组(空白对照组):野生型C57BL/6小鼠;B组(自噬抑制剂对照组):野生型C57BL/6小鼠+自噬抑制剂(3-MA);C组(假手术组1):野生型C57BL/6小鼠+假手术+自噬抑制剂(3-MA);D组(造模组1):野生型C57BL/6小鼠+造模+自噬抑制剂(3-MA);E组(假手术组2):野生型C57BL/6小鼠+假手术+TLR4腺病毒抑制+自噬抑制剂(3-MA);F组(造模组2):野生型C57BL/6小鼠+造模+TLR4腺病毒抑制+自噬抑制剂(3-MA);进行脑组织含水量(BWC)和神经功能障碍评分(NDS);Western Blot检测LC3-Ⅱ/Ⅰ比值,TLR4,MyD88和NF-κB蛋白表达水平;ELISA检测细胞因子TNF-α,IL-1β,IL-6水平。结果与C组比较,E组BWC和NDS下降(P<0.05);与D组比较,F组BWC和NDS下降(P<0.05);A,B,C和D组TLR4,MyD88和NF-κB p65蛋白水平无明显变化(P>0.05);与C组比较,E组TLR4,MyD88,NF-κB p65水平,LC3-Ⅱ/Ⅰ比值下降(P<0.05);与D组比较,F组TLR4,MyD88,NF-κB p65水平,LC3-Ⅱ/Ⅰ比值下降(P<0.05);与A组比较,B,C和D组LC3-Ⅱ/Ⅰ比值下降(P<0.05);与A组比较,B,C和D组TNF-α,IL-1β,IL-6水平下降(P<0.05);与C组比较,E组TNF-α,IL-1β,IL-6水平下降(P<0.05);与D组比较,F组TNF-α,IL-1β,IL-6水平下降(P<0.05)。结论抑制C57BL/6小鼠的TLR4及自噬可减轻自噬引起的脑出血炎症损伤。Objective To investigate the effects of TLR4 mediates the autophagy of microglia on inflam- mation in cerebral hemorrhage. Methods A group： C57BL/6 mouse, B group： C57BL/6 mouse ＋ 3-MA, C group： C57BL/6 mouse ＋ sham operation ＋ 3-MA, D group： C57BL/6 mouse ＋ molding ＋ 3-MA, E group： C57BL/6 mouse ＋ sham operation ＋ TLR4 adenovirus inhibition ＋ 3-MA, F group： C57BL/6 mouse ＋ molding ＋ TLR4 adenovirus inhibition ＋ 3-MA. The water content of brain （BWC） and neurologic deficit score （NDS） were tested in each group. Western blot was used to detect the expression levels of LC3-Ⅱ/Ⅰ rate, TLR4, MyD88 and NF-κB p65. ELISA was used to detect the expression levels of TNF-α, IL-1β, IL-6. Results Compared with C group, the BWC and NDS in E group were lower（P〈0. 05）. Compared with D group, the BWC and NDS in F group were lower（P〈0. 05）. There were no differences on the TLR4, MyD88 and NF-κB p65 protein levels in A, B, C and D group（P〈0. 05）. Compared with C group, the TLR4, MyD88 and NF-κB p65 protein levels, LC3-Ⅱ/Ⅰ rate in E group were lower（P〈0. 05）. Compared with D group, the TLR4, MyD88 and NF-κB p65 protein levels, LC3-Ⅱ/Ⅰ rate in F group were lower（P〈0. 05）. Compared with A group, the LC3-Ⅱ/Ⅰ rate in B, C, D group was lower（P〈0. 05）. Compared with A group, the TNF-α, IL-1βand IL-6 levels in B, C, D group were lower（P〈0. 05）. Compared with C group, the TNF-α, IL-1β and IL-6 levels in E group were lower（P〈0. 05）. Compared with D group, the TNF-α, IL-1β and IL-6 levels in F group were lower（P〈0. 05）. Conclusion Inhibiting TLR4 and autophagy in C57BL/6 could reduce inflamma- tory injury in cerebral hemorrhage.

关 键 词：TLR4 脑出血 炎症 

分 类 号：R743.34[医药卫生—神经病学与精神病学]