沉默TRPM7对无镁外液致痫Neuro-2a细胞炎症反应的影响  

The effect of silencing TRPM7 on the inflammatory response of epileptic Neuro-2a cells induced by magnesium free extracellular fluid

作　　者：王娟 李心雨 杨雪 杨楠 王玉雪 陶怡 朱敬汝 张莉 WANG Juan;LI Xinyu;YANG Xue;YANG Nan;WANG Yuxue;TAO Yi;ZHU Jingru;ZHANG Li(Department of Histology and Embryology of Jinzhou Medical University,Jinzhou 121001,China;Medical Quality Control Office of Jinzhou Traditional Chinese Medicine Hospital,Jinzhou 121001,China)

机构地区：[1]锦州医科大学组织胚胎学教研室 [2]锦州市中医医院医疗质量控制办公室

出　　处：《中国体视学与图像分析》2024年第2期160-166,共7页Chinese Journal of Stereology and Image Analysis

基　　金：辽宁省“双一流”建设项目(30320191022);辽宁省大学生创新训练项目(20201016028);辽宁省教育厅项目(LJKZ0822)。

摘　　要：目的观察沉默瞬时受体电位M7通道(transient receptor potential melastatin 7,TRPM7)对无镁外液致痫小鼠脑神经瘤Neuro-2a细胞炎症反应的影响。方法Neuro-2a细胞经脂质体转染TRPM7 siRNA,经无镁外液培养3 h建立癫痫细胞模型。实验分为4组:对照组(Control)、癫痫组(EP)、癫痫转染组(EP+si-TRPM7)和癫痫转染阴性对照组(EP+si-NC)。成模后24 h,利用实时荧光定量PCR技术检测TRPM7 mRNA表达;利用Western Blot检测HMGB1和TLR4蛋白的表达;利用ELISA方法检测上清液中TNF-α、IL-1β和IL-6的含量;CCK-8法检测细胞活力。结果成模后24 h,与Control组比较,EP组TRPM7 mRNA、HMGB1和TLR4蛋白表达增多,上清液中TNF-α、IL-1β和IL-6含量增多,细胞活力降低。与EP+si-NC组比较,EP+si-TRPM7组TRPM7 mRNA、HMGB1和TLR4蛋白表达减少,上清液中TNF-α、IL-1β和IL-6含量减少,细胞活力增高(P<0.01)。结论沉默TRPM7能抑制HMGB1/TLR4通路,减轻炎症反应,对无镁外液致痫细胞有保护作用。Objective To observe the effect of transient receptor potential M7 channel(TRPM7)gene silencing on the inflammatory response in epileptic Neuro-2a cells induced by magnesium free extracellular fluidMethods The mouse brain neuroma cells(Neuro-2a)were transfected by TRPM7 siRNA through liposomesCells were cultured in magnesium free extracellular medium for 3 hours to establish an epilepsy cell modelThe experiment was divided into four groups:control group(Control),epilepsy group(EP),epilepsy transfection group(EP+si-TRPM7),and epilepsy transfection negative control group(EP+si-NC)The expression of TRPM7 mRNA was detected by real-time fluorescence quantitative PCR technology 24 hours after EP model was builtThe changes of HMGB1 and TLR4 protein were detected by Western BlotELISA measures were used to check the expression of TNF-α,IL-1βand IL-6 in the cell supernatantCCK-8 method was used to detect the cell viabilityResults Compared with the Control group,the expression of TRPM7 mRNA,HMGB1 and TLR4 protein in cells increased in EP group 24 hours after EP model was builtThe expression of TNF-α,IL-1βand IL-6 in the cell supernatant increased tooWhile the cell viability decreasedCompared with the EP+si-NC group,the expression of TRPM7 mRNA,HMGB1 and TLR4 protein in cells decreased in EP+si-TRPM7 groupThe expression of TNF-α,IL-1βand IL-6 in the cell supernatant decreased tooWhile the cell viability increased(P<001)Conclusions Silencing TRPM7 can inhibit the HMGB1/TLR4 pathway,alleviate inflammatory response,and have a protective effect on epileptic cells induced by magnesium free extracellular fluid.

关 键 词：瞬时受体电位M7通道 癫痫 无镁外液 Neuro-2a细胞 炎症反应 

分 类 号：R742.1[医药卫生—神经病学与精神病学] N3[医药卫生—临床医学]