Mito-TEMPO通过减轻线粒体损伤抑制铅诱导的BV2小胶质细胞活化
Mito-TEMPO inhibits the activation of BV2 microglia induced by lead exposure by reducing mitochondrial damage
陈超 1赵再华 2王涛 2卢金锁 3郑刚2
作者信息
- 1. 西安建筑科技大学环境与市政工程学院,西安 710055;空军军医大学军事预防医学系军队劳动与环境卫生学教研室,特殊作业环境危害评估与防治教育部重点实验室,西安 710032
- 2. 空军军医大学军事预防医学系军队劳动与环境卫生学教研室,特殊作业环境危害评估与防治教育部重点实验室,西安 710032
- 3. 西安建筑科技大学环境与市政工程学院,西安 710055
- 折叠
摘要
目的:研究线粒体氧化应激在铅暴露诱导的小胶质细胞活化中的作用,观察靶向线粒体的抗氧化剂Mito-TEMPO的保护作用.方法:培养小鼠小胶质细胞系(BV2),建立铅暴露模型.采用噻唑蓝(MTT)比色法测定铅暴露对细胞活力的影响,采用免疫荧光染色法检测M1型活化标志物CD86蛋白的表达;采用酶联免疫吸附试验(ELISA)检测白细胞介素-1β(IL-1β)、肿瘤坏死因子-α(TNF-α)、白细胞介素-6(IL-6)的含量;采用线粒体超氧化物(MitoSOX)染色检测线粒体氧化应激水平;采用线粒体膜电位荧光探针(JC-1)染色检测线粒体膜电位;采用细胞能量代谢分析(O2k)检测线粒体呼吸能力.采用Mito-TEMPO抗氧化剂进行干预,研究Mito-TEM-PO对BV2线粒体氧化应激、细胞活化的抑制作用.结果:与对照组相比,铅暴露组BV2细胞CD86蛋白表达水平显著升高,促炎因子IL-1β、TNF-α、IL-6水平显著升高,线粒体氧化应激水平显著升高,线粒体膜电位、呼吸能力显著降低.Mito-TEMPO处理显著减轻了 BV2细胞线粒体氧化应激与功能损伤,并显著抑制了 BV2细胞M1型活化水平.结论:Mito-TEMPO能够逆转铅暴露诱导的BV2细胞M1型活化,其机制可能与Mito-TEMPO减轻线粒体氧化应激与功能损伤有关.
Abstract
Objective:To explore the mechanism of Mito-TEMPO inhibiting the activation of BV2 microglia induced by lead(Pb)exposure.Methods:Mouse microglia BV2 were cultured in vitro.The effects of different concentrations of lead exposure on the viability of BV2 cells were determined by MTT colorimetric method,and a model of lead expo-sure was developed and intervened with Mito-TEMPO antioxidant.Immunofluorescence staining was used to detect the expression of M1 activation marker CD86 protein.Enzyme-linked immunosorbent assay was used to detect the expression of pro-inflammatory factors interleukin-1 β(IL-1β),tumor necrosis factor α(TNF-α),interleukin-6(IL-6).Mito-chondrial superoxide indicator(MitoSOX)staining was used to detect the level of mitochondrial oxidative stress.JC-1 staining was used to detect mitochondrial membrane potential.The respiratory ability of mitochondria was detected by cell energy metabolism analysis system(O2k).Results:Compared with the control group,the expression of CD86 protein,the levels of pro-inflammatory cytokines IL-1β,TNF-α and IL-6 in BV2 cells increased significantly,the level of oxidative stress in mitochondria increased significantly,and the mitochondrial membrane potential and respiratory ability decreased significantly in lead exposed group.Mito-TEMPO treatment significantly reduced the oxidative stress and functional damage of mitochondria in BV2 cells,and significantly inhibited the M1 activation level of BV2 cells.Conclusion:The results show that Mito-TEMPO treatment can reverse the M1 activation of BV2 cells induced by lead exposure,and the specific mechanism may be related to the reduction of mitochondrial oxidative stress and dysfunction by Mito-TEMPO.
关键词
Mito-TEMPO/铅/线粒体/炎症/BV2小胶质细胞Key words
Mito-TEMPO/lead/mitochondria/inflammation/BV2 microglia引用本文复制引用
基金项目
国家自然科学基金重点国际合作与交流项目(81920108030)
出版年
2024
NETL
NSTL
万方数据