下调miR-203靶向MEF2C/NF-κB抑制颞叶癫痫大鼠 海马胶质细胞的活化和炎症反应  

Down-regulation of miR-203 inhibits glial cell activation and inflammation in hippocampal tissues of temporal lobe epilepsy rats by targeted regulation of MEF2C/NF-κB pathway

作　　者：许超 王娟 刘锋 梁洁敏 刘伟伟[2] XU Chao;WANG Juan;LIU Feng;LIANG Jie-min;LIU Wei-wei(Department of Medicine,Changjiang University,Jingzhou 434023,China;Department of Neurosurgery,The First Affiliated Hospital of Changjiang University,Jingzhou 434000,China)

机构地区：[1]长江大学医学部,湖北荆州434023 [2]长江大学附属第一医院神经外科,湖北荆州434000

出　　处：《中国临床神经外科杂志》2021年第6期449-454,共6页Chinese Journal of Clinical Neurosurgery

摘　　要：目的探讨下调miR-203对颞叶癫痫大鼠海马胶质细胞活化和炎症反应的影响。方法将46只成年SD大鼠随机分为假手术组(n=10)、模型组(n=12)、阴性对照组(n=12)和miR-203低表达组(n=12)。立体定向辅助下,将海人酸注入大鼠左侧海马CA3区建立颞叶癫痫模型,miR-203低表达组和阴性对照组大鼠左侧海马CA3区分别注射携带miR-203 inhibitor的腺相关病毒和空腺病毒载体,7 d后取左侧海马组织,PCR法检测miR-203水平,ELISA法检测炎症因子[白细胞介素-1β(IL-1β)、IL-6和肿瘤坏死因子α(TNF-α)]水平,免疫印迹法检测肌细胞增强因子2c(MEF2C)、核因子-κB(NF-κB)p65蛋白表达,TUNEL法检测神经元凋亡,GFAP/CD11b/c免疫荧光染色分析胶质细胞活化。结果模型组大鼠海马组织miR-203、NF-κB p65蛋白表达量、炎症因子(IL-1β、IL-6和TNF-α)水平明显增高(P<0.05),MEF2C蛋白表达量明显降低(P<0.05),活化的胶质细胞数量、神经元凋亡数量明显增多(P<0.05)。双荧光素酶报告基因实验显示,MEF2C是miR-203的靶基因。miR-203表达低表达组大鼠海马组织miR-203表达量、NF-κB p65蛋白表达量、炎症因子(IL-1β、IL-6和TNF-α)水平明显降低(P<0.05),MEF2C蛋白表达量明显增高(P<0.05),活化的胶质细胞数量、神经元凋亡数量明显减少(P<0.05)。结论下调miR-203,靶向调控MEF2C/NF-κB信号通路,抑制颞叶癫痫大鼠海马胶质细胞的活化、神经元凋亡和炎症反应。Objective To explore the effect of down-regulation of miR-203 on the glial cell activation and inflammation in hippocampal tissues of temporal lobe epilepsy rats.Methods Forty-six adult SD rats were randomly divided into sham operation group(n=10),model group(n=12),negative control group(n=12)and miR-203 low expression group(n=12).Under stereotactic assistance,kainic acid was injected into the left hippocampus CA3 to establish the temporal lobe epilepsy model.Adeno-associated viral(AAV)integrated with miR-203 inhibitor and AAV vector were injected into the left hippocampal CA3of the rats in the miR-203 low expression group and the negative control group,respectively.Seven days after the intervention,the left hippocampal tissues were isolated,and the level of miR-203 was detected by PCR,and the inflammatory factors including interleukin-1β(IL-1β),IL-6 and tumor necrosis factorα(TNF-α)were detected by ELISA,the expression levels of myocyte enhancer factor 2c(MEF2C),nuclear factorκB(NF-κB)p65 protein were detected by immunoblotting method,neuronal apoptosis was detected by TUNEL method,and glial cell activation was detected by GFAP/CD11b/c immunofluorescence staining.Results In the hippocampal tissues of rats in model group,the expression levels of miR-203 and NF-κB p65 protein,and the levels of inflammatory factors(IL-1β,IL-6 and TNF-α)were significantly increased(P<0.05),and the expression level of MEF2C protein was significantly decreased(P<0.05),the numbers of activated glial cells and apoptic neurons were significantly increased(P<0.05).The dual luciferase reporter gene experiment showed that MEF2C was the target gene of miR-203.In the hippocampal tissues of rats in miR-203 low expression group,the expression levels of miR-203 and NF-κB p65 protein,and the levels of inflammatory factors(IL-1β,IL-6 and TNF-α)were significantly reduced(P<0.05),the protein expression level of MEF2C was significantly increased(P<0.05),and the numbers of activated glial cells and apoptic neurons were significantly reduce

关 键 词：颞叶癫痫 miR-203 MEF2C/NF-κB通路 胶质细胞活化 炎症反应 

分 类 号：R742.1[医药卫生—神经病学与精神病学] Q786[医药卫生—临床医学]