摘要
目的探讨小胶质细胞及星形胶质细胞中的瞬时受体电位M2型(TRPM2)离子通道在颞叶癫痫(TLE)相关神经炎症中的作用。方法将大鼠随机分为对照组和癫痫组,癫痫组采用氯化锂-毛果芸香碱(匹罗卡品)制作癫痫模型,对照组给予等剂量的生理盐水代替,根据造模后观察的时间将TLE大鼠随机分为7个亚组(n=5):急性期(3 h组、6 h组、1 d组、2 d组)、潜伏期(14 d组)、慢性期(30 d组、90 d组)。检测TRPM2在不同亚组TLE大鼠海马中的表达及TRPM2在大鼠海马中的细胞定位情况。采用过氧化氢(H_(2)O_(2))诱导小胶质细胞BV2及星形胶质细胞C8细胞系,检测细胞释放IL-1β、IL-6和TNF-α的水平,检测H_(2)O_(2)诱导的BV2及C8细胞中TRPM2、聚腺苷二磷酸核糖聚合酶-1(PARP-1)、糖原合成酶激酶3β(GSK-3β)、磷酸化核因子κB p65蛋白(p-NF-κB p65)的表达变化,同时检测H_(2)O_(2)诱导BV2细胞中钙调神经磷酸酶(CaN)的活性变化。结果急性期TLE大鼠海马中的TRPM2表达升高(P<0.05)。H_(2)O_(2)诱导的BV2及C8细胞中TRPM2的表达升高、炎症因子释放增加(P均<0.05),H_(2)O_(2)可促进BV2细胞中PARP-1、p-NF-κB p65的表达并提高CaN、GSK-3β的活性(P均<0.05)。结论氧化应激可促进小胶质细胞及星形胶质细胞TRPM2及促炎细胞因子的表达,癫痫反复发作引起的氧化应激可能在小胶质细胞中通过TRPM2/CaN/p-NF-κB p65通路介导TLE相关神经炎症的发生。
Objective To investigate the role of transient receptor potential melatonin 2(TRPM2)ion channels in microglia and astrocytes in temporal lobe epilepsy(TLE)-associated neuroinflammation.Methods Rats were randomly divided into the control group and epilepsy group.Seizure models were induced by lithium chloride-pilocarpine in the epilepsy group,and those in the control group were injected with the same dose of saline.The TLE rats were randomly divided into 7 subgroups according to the observation time after model establishment:acute phase(3 h group,6 h group,1 d group,2 d group),latent phase(14 d group)and chronic phase(30 d group,90 d group)subgroups(n=5).The expression level of TRPM2 in the hippocampus of TLE rats at different stages were detected,and the cellular localization of TRPM2 in the brain of TLE rats was investigated.BV2 and C8 cell lines were induced by hydrogen peroxide(H_(2)O_(2)).The levels of IL1-β,IL-6 and TNF-αreleased by BV2 and C8 cells were observed by ELISA.The levels of TRPM2,poly(ADP-ribose)polymerase 1(PARP-1),glycogen synthase kinase-3β(GSK-3β)and phosphorylated nuclear factor-κb p65(p-NF-κb p65)were measured in H_(2)O_(2)-induced BV2 and C8 cells.Meantime,the activity of Calcineurin(CaN)in H_(2)O_(2)-induced BV2 was observed.Results The TRPM2 expression in 2d epileptic rat hippocampus was increased(P<0.05).The expression of TRPM2 and the release of inflammatory cytokines were increased in H_(2)O_(2)induced-BV2 and C8 cells(both P<0.05).H_(2)O_(2)could up-regulate the expression of PARP-1 and p-NF-κB p65 and enhance the activity of CaN in BV2 cells(all P<0.05).Conclusions Oxidative stress can up-regulate the expression levels of TRPM2 and pro-inflammatory cytokines in microglia and astrocytes.Oxidative stress caused by recurrent epilepsy may mediate the occurrence of TLE-associated neuroinflammation through the TRPM2/CaN/p-NF-κB p65 pathway in microglia.
作者
王星辰
刘瑞寒
肖翔宇
夏敏
李秋波
孔庆霞
Wang Xingchen;Liu Ruihan;Xiao Xiangyu;Xia Min;Li Qiubo;Kong Qingxia(Cheeloo College of Medicine,Shandong University,Jinan 250012,China)
出处
《新医学》
CAS
2024年第3期214-221,共8页
Journal of New Medicine
基金
济宁市重点研发计划(2021YXNS092)
济宁医学院贺林院士新医学临床转化工作站科研基金(JYHL2019FMS25)。
