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烟酰胺核糖在视神经节细胞中的线粒体保护作用

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目的 探究烟酰胺核糖(NR)在羰基氰化物间氯苯腙(CCCP)所诱导的R28细胞氧化应激损伤模型中的保护作用。方法 使用4 μmol/L CCCP诱导R28细胞产生氧化应激,并使用400 nmol/L NR进行干预,CCK-8实验检测细胞活力,Annexin-V/PI双染流式细胞术检测细胞凋亡,蛋白质印记法检测细胞色素C、半胱氨酸蛋白酶-3、半胱氨酸蛋白酶-9表达水平以评估细胞凋亡;同时用四甲基罗丹明乙酯检测线粒体膜电位(MMP),MitoSOX检测线粒体活性氧(mtROS)水平,三磷酸腺苷(ATP)试剂盒检测ATP生成能力以评价线粒体功能。结果 CCCP处理R28细胞后,细胞活力下降,凋亡蛋白水平和凋亡率上升,MMP下降,mtROS生成增加(P<0。05);NR预处理后,细胞活力上升,凋亡蛋白水平和细胞凋亡率下降,MMP上升,mtROS生成减少(P<0。05)。结论 NR通过抑制氧化应激反应,保护线粒体功能,从而增强细胞活性,降低凋亡蛋白的表达,最终减少视网膜神经节细胞凋亡。
The protective effect of nicotinamide riboside on mitochondrial function of retinal ganglion cell
Objective To explore the protective effect in a model of nicotinamide riboside(NR)against carbonyl cyanide m-chlorophenylhydrazone(CCCP)-induced oxidative stress in R28 cells.Methods 4 μmol/L CCCP was used to induce oxidative stress in R28 cells,and 400 nmol/L NR was used to intervene.The cell viability was quantified by CCK-8 assay.The apoptosis was detected by Annexin-V/PI double staining and flow cytometry.Western blotting was used to examine the levels of Cytochrome C,Caspase-3,and Caspase-9 to evaluate the apoptosis.Tetramethylrhodamine ethyl ester was used to detect the mitochondrial membrane potential(MMP),MitoSOX was used to detect the mitochondrial reactive oxygen species(mtROS)levels,and adenosine triphosphate(ATP)assay kit was used to assess ATP generation ability to evaluate mitochondrial function.Results After CCCP treatment of R28 cells,the cell viability decreased,the apoptotic protein levels and apoptosis rates increased,the MMP decreased,and the mtROS generation increased(P<0.05).After NR pretreatment,the cell viability increased,the apoptotic protein levels and apoptosis rates decreased,the MMP increased,and the mtROS generation decreased(P<0.05).Conclusion:NR enhances the cell viability,reduces the expression of apoptotic proteins,and ultimately reduces the apoptosis of retinal ganglion cell by inhibiting oxidative stress response and protecting mitochondrial function.

Nicotinamide ribosideMitochondriaOxidative stressOptic nerve protection

邓翕之、张楠、曾雯、朱敏、张鹏宇、李芳、蒋斌、柯敏

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武汉大学中南医院眼科(武汉 430071)

烟酰胺核糖 线粒体 氧化应激 视神经保护

国家自然科学基金面上项目中国博士后科学基金第72批面上资助项目

819708022022M722463

2024

中国药师
国家药品监督管理局高级研修学院,武汉医药(集团)股份有限公司

中国药师

CSTPCD
影响因子:0.944
ISSN:1008-049X
年,卷(期):2024.27(1)
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