瑞马唑仑通过调节Toll样受体4对脂多糖诱导的小胶质细胞M1型极化的影响
Effect of remimazolam on LPS-induced microglial M1 polarization by regulating TLR4
姜丰 1李婧 2刘文洁 3张高峰 3袁阳 3时飞3
作者信息
- 1. 266033 青岛大学附属青岛市海慈医院(青岛市中医医院)麻醉科
- 2. 青岛市市立医院妇科
- 3. 青岛市市立医院麻醉科
- 折叠
摘要
目的 评价瑞马唑仑在脂多糖诱导的M1型小胶质细胞极化中的作用及其与Toll样受体4(Toll-like recep-tor 4,TLR4)的关系.方法 将生长良好的BV2小胶质细胞采用随机数字表法分为5组(n=20):对照组、脂多糖组、瑞马唑仑+脂多糖组(瑞马唑仑组)、瑞马唑仑+脂多糖+Neoseptin-3组(激动剂组)、瑞马唑仑+脂多糖+二甲基亚砜组(激动剂对照组).对照组置于正常条件下培养,脂多糖组加入浓度为1 μg/ml的脂多糖孵育24 h,瑞马唑仑组经100 μg/ml瑞马唑仑预处理20 min,激动剂组和激动剂对照组分别给予TLR4激动剂Neoseptin-350 μmol(用二甲基亚砜溶解)和等容量二甲基亚砜孵育1 h.采用酶联免疫吸附测定法检测细胞上清液肿瘤坏死因子α(tumor necrosis factor alpha,TNF-α)、白细胞介素(interleukin,IL)-1β水平,采用定量逆转录聚合酶链反应法检测M1型小胶质细胞标志物诱导型一氧化氮合酶(inducible nitric oxide synthase,iNOS)信使核糖核酸(messen-ger RNA,mRNA)和TLR4 mRNA表达水平.结果 与对照组比较,其余4组细胞上清液TNF-α、IL-1β、iNOS和TLR4蛋白及其mRNA表达明显升高(P＜0.05).与脂多糖组比较,瑞马唑仑组细胞上清液TNF-α、IL-1β、iNOS和TLR4蛋白及其mRNA表达明显降低(P＜0.05).与瑞马唑仑组比较,激动剂组细胞上清液TNF-α、IL-1β水平、iNOS和TLR4蛋白及其mRNA表达明显升高(P＜0.05).激动剂对照组上清液TNF-α、IL-1β 水平、iNOS和TLR4蛋白及其mRNA与瑞马唑仑组比较,差异无统计学意义(P＞0.05).脂多糖组iNOS表达明显高于对照组[(14.757±0.986)％vs(1.561±0.08)％,P＜0.05].瑞马唑仑组 iNOS 表达明显低于脂多糖组[(3.767±0.364)％vs(14.757±0.986)％,P＜0.05].激动剂组 iNOS 表达高于瑞马唑仑组[(6.827±0.642)％vs(3.767±0.364)％,P＜0.05].激动剂对照组与瑞马唑仑组iNOS表达比较,无统计学差异(P＞0.05).结论 瑞马唑仑抑制脂多糖诱导的M1型小胶质细胞极化的机制与下调TLR4的表达有关.
Abstract
Objective To evaluate the role of remimazolam in lipopolysaccharide(LPS)-induced M1 microglial polarization and its relationship with Toll-like receptor 4(TLR4).Methods BV2 mi-croglia cells were randomly divided into 5 groups(n=20):control group,LPS group(1 µg/ml for 24 h),remimazolam+LPS group(remimazolam group,pretreated with 100 μg/ml remimazolam for 20 min followed by LPS),remimazolam+LPS+Neoseptin-3 group(agonist group,50 pmol Neoseptin-3 dissolved in DMSO),and remimazolam+LPS+DMSO group(agonist control group).The contents of TNF-α and IL-1β in the supernatant were detected by ELISA.The expres-sion of M1 microglia markers,inducible nitric oxide synthase(iNOS)and TLR4 at mRNA.Immu-nofluorescence staining was employed to identify the location of iNOS.Results When compared to the control group,the contents of TNF-α and IL-1β in the supernatant and the expression of iNOS[(14.757±0.986)％vs(1.561±0.08)％]and TLR4 at mRNA and protein levels were sig-nificantly higher in the other four groups(P＜0.05).Remimazolam treatment reversed the increa-ses of the TNF-α and IL-1β contents in the supernatant and mRNA and protein expression of iNOS[(3.767±0.364)％vs(14.757±0.986)％]and TLR4 induced by LPS(P＜0.05).In addi-tion,remimazolam agonist Neoseptin-3 restored the effects of LPS on above molecules[iNOS:(6.827±0.642)％vs(3.767±0.364)％,all P＜0.05].But,there were no statistical differences in above molecules between the agonist group and agonist control group(P＞0.05).Conclusion The mechanism by which remimazolam inhibits LPS-induced M1 microglial polarization is related to down-regulation of TLR4 expression.
关键词
小神经胶质细胞/苯二氮卓类药物/脂多糖类/Toll样受体4Key words
microglia/benzodiazepines/lipopolysaccharides/Toll-like receptor 4引用本文复制引用
出版年
2025
NETL
NSTL
万方数据