摘要
目的探讨线粒体钙离子单向转运体在β淀粉样蛋白诱导小胶质细胞凋亡中的作用机制。方法将体外原代培养小胶质细胞随机分为对照组、Aβ_(25-35)组、Ru360组、Spermine组,并进一步采用线粒体特异性抗氧化剂MitoQ进行干预;采用流式细胞术、荧光探针及Western blotting等技术检测细胞凋亡、线粒体钙离子浓度、ROS生成及相关蛋白表达。结果 (1)与对照组相比,Aβ_(25-35)显著增加小胶质细胞凋亡,Ru360显著减少Aβ_(25-35)诱导的小胶质细胞凋亡,而Spermine发挥相反作用;(2)与对照组相比,Aβ_(25-35)显著增加低线粒体钙离子浓度及线粒体ROS生成水平,Ru360显著降低Aβ_(25-35)诱导的线粒体钙离子浓度及线粒体ROS生成水平的升高,而Spermine发挥相反作用;(3)与对照组相比,Aβ_(25-35)组GRP78、CHOP和caspase-12的表达显著增加,Ru360显著减少Aβ_(25-35)诱导的GRP78、CHOP和caspase-12表达的增加,而Spermine发挥相反作用;(4)与Aβ_(25-35)组相比,MitoQ组显著减少线粒体ROS产物水平,并减少GRP78、CHOP和caspase-12表达。结论 (1) MCU在Aβ_(25-35)诱导的小胶质细胞凋亡发挥重要作用,抑制MCU可减少细胞凋亡,而激活MCU增强细胞凋亡;(2)氧化应激介导的内质网应激反应可能在MCU参与Aβ_(25-35)诱导的小胶质细胞凋亡中发挥重要作用。
Objective To investigate the effect and mechanism of mitochondrial calcium uniporter on β-amyloid-induced endoplasmic reticulum stress and apoptosis in microglia.Methods Primary microglial cells were randomly divided into control group,Aβ 25-35 group,Ru360 group and Spermine group.In addition,mitochondrial specific antioxidant MitoQ was adopted.We detected apoptotic cells,mitochondrial calcium concentrations,ROS production and apoptotic proteins by flow cytometry analysis,fluorescent probes and Western blotting.Results (1)Compared with control group,Aβ 25-35 significantly increased microglial apoptosis;Pretreatment with Ru360 significantly decreased Aβ 25-35 -induced microglial apoptosis,while Spermine exerted the opposite effect.(2)Compared with control group,Aβ 25-35 significantly increased mitochondrial calcium ion concentration and mitochondrial ROS production levels;Pretreatment with Ru360 significantly reduced mitochondrial calcium ion concentration and mitochondrial ROS production levels,while Spermine exerted the opposite effect.(3)Compared with control group,Aβ 25-35 significantly increased the expressions of CHOP,GRP78 and caspase-12;Pretreatment with Ru360 significantly reduced the expressions of CHOP,GRP78 and caspase-12,while Spermine exerted the opposite effect.(4)Compared with Aβ 25-35 group,MitoQ significantly reduced mitochondrial ROS production and the expressions of CHOP,GRP78 and caspase-12.Conclusion (1)Inhibition of MCU exerts neuroprotective effects on Aβ-induced microglia apoptosis.(2)Oxidative stress mediated endoplasmic reticulum stress may play an important role in this process.
作者
谢南昌
余梦嫣
王翠
李英娇
孟祥荷
连亚军
XIE Nanchang;YU Mengyan;WANG Cui(Department of Neurology,The First Affiliated Hospital of Zhengzhou University,Zhengzhou 450000,China)
出处
《中风与神经疾病杂志》
CAS
2019年第2期99-103,共5页
Journal of Apoplexy and Nervous Diseases
基金
国家自然科学基金面上项目(No.81571260)
