山西大同大学学报(自然科学版)2024,Vol.40Issue(1) :75-80.DOI:10.3969/j.issn.1674-0874.2024.01.014

雷公藤甲素通过调控NLRP3通路抑制LPS诱导的BV2小胶质细胞炎性反应

Triptolide Inhibits LPS-induced Inflammatory Response in BV2 Microglia by Modulating the NLRP3 Pathway

穆秉桃 李玉璐 刘淇源 孙肖乐 胡芬琦 曹佳蕾 杨凤君 李佩佩 毕宇锦 王泽涛 张慧宇
山西大同大学学报(自然科学版)2024,Vol.40Issue(1) :75-80.DOI:10.3969/j.issn.1674-0874.2024.01.014

雷公藤甲素通过调控NLRP3通路抑制LPS诱导的BV2小胶质细胞炎性反应

Triptolide Inhibits LPS-induced Inflammatory Response in BV2 Microglia by Modulating the NLRP3 Pathway

穆秉桃 1李玉璐 2刘淇源 2孙肖乐 2胡芬琦 2曹佳蕾 2杨凤君 2李佩佩 2毕宇锦 2王泽涛 2张慧宇3
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作者信息

  • 1. 山西大同大学脑科学研究所/分子细胞免疫学大同市重点实验室,山西大同 037009;山西大同大学医学院,山西大同 037009
  • 2. 山西大同大学医学院,山西大同 037009
  • 3. 山西大同大学脑科学研究所/分子细胞免疫学大同市重点实验室,山西大同 037009;山西大同大学中医药健康服务学院,山西大同 037009
  • 折叠

摘要

目的 基于炎症小体(NLRP3)通路探讨雷公藤甲素(Triptolide TP)抑制小胶质细胞炎性反应的机制.方法 脂多糖(LPS)刺激BV2小胶质细胞促细胞炎症损伤,建立体外模型,观察TP对LPS刺激的小胶质细胞的影响.BV2小胶质细胞分为对照组、LPS诱导的模型组(1 mg/L LPS)、TP组(1 nmol/L TP+1 mg/L LPS).对照组不做处理,其它组药物作用24 h,镜下观察药物组细胞发生的形态变化并和对照组比较;细胞上清液用Griess法测定一氧化氮(NO)的释放量;免疫荧光染色和Western blot法检测各组细胞间炎症小体蛋白3(NLRP3)、凋亡相关斑点样蛋白(ASC)、半胱氨酸天冬氨酸蛋白酶(caspase-1)和白细胞介素-18(IL-18)的水平差别.结果 LPS诱导BV2细胞炎性活化,形态呈阿米巴样,降低细胞活性,促进NO的释放.TP作用后降低NO水平,使NLRP3炎性小体激活受到抑制,NLRP3、ASC、caspase-1和IL-18表达水平低下.结论 TP对小胶质细胞炎性反应有抑制作用,发挥了TP的抗炎作用,抑制了NLRP3炎性小体的活化和表达.

Abstract

Objective To investigate the mechanisation of triptolide TP inhibiting microglial inflammation based on the inflam-mosome(NLRP3)pathway.Methods BV2 microglia and lipopolysaccharide(LPS)were selected to stimulate microglia to promote cell inflammatory damage,and an in vitro model was established to observe the effect of TP on LPS-stimulated microglia.The BV2 microglia was divided into three experimental groups:control group,LPS-induced model group(1 μg/mL LPS),and TP group(1 nmol/L TP+1 μg/mL LPS).The control group was not treated,the other groups was treated by the drug for 24 h,then the morpholog-ical changes of the cells in the drug groups were observed by microscope and compared with the control group.Cell supernatants are determined by the Griess method for nitric oxide(NO)release;Immunofluorescence staining and Western blot assays were used to detect the levels of intercellular inflamamite protein 3(NLRP3),apoptosis-associated spot-like protein(ASC),cysteine aspartate pro-tease(caspase-1),and interleukin-18(IL-18).Results LPS-induced inflammatory activation of BV2 cells,which was amoebias-like,reduced cell activity and promoted the release of NO.After TP,it reduces NO levels,inhibits the activation of NLRP3 inflamma-somes,and the expression of the corresponding components NLRP3,ASC,caspase1 and IL-18 was dropped.Conclusion TP has an inhibitory effect on microglial inflammation,exerting its anti-inflammatory effect and inhibiting the activation and expression of NLRP3 inflammasomes.

关键词

雷公藤甲素/脂多糖/BV2细胞/NLRP3炎症小体/炎症反应

Key words

triptolide/lipopolysaccharides/BV2 cells/NLRP3 inflammasome/inflammatory response

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基金项目

山西大同大学大学生创新创业训练计划项目(XDC2022186)

山西省基础研究计划项目(20210302123478)

出版年

2024
山西大同大学学报(自然科学版)
山西大同大学

山西大同大学学报(自然科学版)

影响因子:0.271
ISSN:1674-0874
参考文献量1
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