没食子酸通过NF-κB通路对缺氧诱导的BV2小胶质细胞损伤的保护作用机制
Gallic Acid in Improving Hypoxia-induced BV2 Microglia Injury via Inhibiting NF-κB Pathway
陈洁 1陈科 1孟宪丽 2王小博 2陈小睿1
作者信息
- 1. 成都中医药大学 药学院,四川 成都 611137
- 2. 成都中医药大学 创新研究院/交叉学科研究院,四川 成都 611137
- 折叠
摘要
目的:建立BV2 细胞缺氧损伤模型,研究没食子酸通过NF-κB通路对缺氧诱导的BV2 小胶质细胞损伤的保护作用机制.方法:取BV2 小胶质细胞分为空白组、模型组、低剂量、中剂量及高剂量组,通过三气培养箱建立BV2 小胶质细胞缺氧损伤模型,低剂量、中剂量及高剂量组分别给予 1.7 g/mL、3.4 g/mL、8.5 g/mL没食子酸进行干预.采用CCK-8 法检测没食子酸对缺氧的BV2 细胞的保护作用,使用试剂盒检测上清中乳酸脱氢酶(LDH)、细胞中超氧化物歧化酶(SOD)活力、丙二醛(MDA)和活性氧(ROS)的含量,ELISA检测上清中炎症因子表达水平(TNF-α、IL-1β、IL-6),Western blot检测BV2 细胞内p-IKK/IKK、p-IκB/IκB和pNF-κB p65/NF-κB p65 蛋白的表达水平以及NF-κB p65 的核位移.结果:与空白对照组相比,缺氧可造成BV2 细胞培养基上清液LDH的活力增加和炎症因子(TNF-α、IL-1 β、IL-6)的释放,细胞中SOD活力降低,MDA和ROS的水平升高,NF-κB 通路(p-IKK/IKK、p-IκB/IκB 和p-NF-κB p65/NF-κB p65 蛋白的表达水平)的激活以及NF-κB p65 的核位移.没食子酸能通过降低LDH的活力,抑制氧化应激(提高SOD的活力,下调MDA和ROS水平)、抑制炎症因子的释放、抑制NF-κB通路相关蛋白的激活以及NF-κB p65 的核位移改善缺氧引起的BV2 细胞损伤.结论:没食子酸对缺氧诱导的BV2 细胞炎症具有保护作用.
Abstract
Objective:To establish hypoxia injury model of BV2 microglia,investigating the gallic acid in improving hypoxia-induced BV2 microglia injury.Methods:BV2 microglia were divided into blank group,model group,low-dose,medium-dose and high-dose groups.Hypoxia injury model of BV2 microglia was established in a three-gas incubator.Low-dose,medium-dose and high-dose groups were treated with gallic acid of 1.7 g/mL、3.4 g/mL、8.5 g/mL.CCK-8 method was use to detected the protective effect of gallic acid on hypoxia-induced BV2 microglia injury.LDH viability in supernatant,SOD activity,MDA and ROS content in cells were detected using the kit.Inflammatory factors(TNF-α、IL-1β、IL-6)in supernatant were detected by ELISA kits.The expression level of p-IKK/IKK,p-IκB/IκB and p-NF-κB p65/NF-κB p65 and nuclear displacement of NF-κB p65 in BV2 microglia were detected by Western blot.Results:Compared with control group,hypoxia can increase LDH activity and inflammatory factors(TNF-αIL-1βIL-6)in superna-tant,decrease SOD activity and increased MDA and ROS content in in cells,and NF-κB pathway(P-IKK/IKK P-I κB/IκB and P-NF-κB P65/NF-κB p65 protein expression level)and nuclear displacement of NF-κB p65.Gallic acid can improve hypoxia-induced BV2 microglia injury via inhibiting oxidative stress(Increase SOD activity,decrease MDA and ROS content)and the release of inflammatory factors,inhibiting the activation of NF-κB pathway and nuclear displacement of NF-κB p65.Conclusion:Gallic acid has a protective effect on hypoxia-induced BV2 microglia injury.
关键词
没食子酸/缺氧/BV2/小胶质细胞/炎症/NF-κB通路Key words
Gallic acid/Hypoxia/BV2 microglia/Inflammation/NF-κB pathway引用本文复制引用
基金项目
国家自然科学基金(81973569)
国家自然科学基金(82274207)
国家自然科学基金(82104533)
出版年
2024
国家科技期刊平台
NSTL
万方数据