中国体视学与图像分析2024,Vol.29Issue(2) :160-166.DOI:10.13505/j.1007-1482.2024.29.02.008

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

王娟 李心雨 杨雪 杨楠 王玉雪 陶怡 朱敬汝 张莉
中国体视学与图像分析2024,Vol.29Issue(2) :160-166.DOI:10.13505/j.1007-1482.2024.29.02.008

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

王娟 1李心雨 2杨雪 2杨楠 2王玉雪 2陶怡 2朱敬汝 2张莉2
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作者信息

  • 1. 锦州医科大学组织胚胎学教研室,锦州 121001;锦州市中医医院医疗质量控制办公室,锦州 121000
  • 2. 锦州医科大学组织胚胎学教研室,锦州 121001
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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TRPM7mRNA、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.

Abstract

Objective To observe the effect of transient receptor potential M7 channel(TRPM7)gene si-lencing on the inflammatory response in epileptic Neuro-2a cells induced by magnesium free extracellular fluid.Methods The mouse brain neuroma cells(Neuro-2a)were transfected by TRPM7 siRNA through liposomes.Cells were cultured in magnesium free extracellular medium for 3 hours to establish an epilepsy cell model.The 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 technol-ogy 24 hours after EP model was built.The changes of HMGB1 and TLR4 protein were detected by West-ern Blot.ELISA measures were used to check the expression of TNF-α,IL-1β and IL-6 in the cell su-pernatant.CCK-8 method was used to detect the cell viability.Results 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 built.The expression of TNF-α,IL-1 β and IL-6 in the cell supernatant in-creased too.While the cell viability decreased.Compared with the EP+si-NC group,the expression of TRPM7 mRNA,HMGB1 and TLR4 protein in cells decreased in EP+si-TRPM7 group.The expression of TNF-α,IL-1β and IL-6 in the cell supernatant decreased too.While the cell viability increased(P<0.01).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

Key words

transient receptor potential melastatin 7/epilepsy/magnesium free extracellular medium/Neuro-2a cell/inflammatory response

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基金项目

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)

出版年

2024
中国体视学与图像分析
中国体视学学会

中国体视学与图像分析

影响因子:0.293
ISSN:1007-1482
参考文献量3
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