Protective Effects of Nicotinamide Riboside on H2O2-induced Oxidative Damage in Lens Epithelial Cells
Protective Effects of Nicotinamide Riboside on H2O2-induced Oxidative Damage in Lens Epithelial Cells
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DOI：
10.1080/02713683.2020.1855662
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发表时间：
2020-12
影响因子：
2
通讯作者：
Bi-ting Zhou;Guangyu Zhao;Yihua Zhu;Xiaole Chen;Nanwen Zhang;Juhua Yang;Hong Lin
中科院分区：
文献类型：
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作者：
Bi-ting Zhou;Guangyu Zhao;Yihua Zhu;Xiaole Chen;Nanwen Zhang;Juhua Yang;Hong Lin
摘要目的：目的探讨烟酰胺核苷（NR）对过氧化氢（H2 O2）诱导的人透镜上皮细胞株（SRA 01/04）氧化损伤的保护作用及其可能机制。材料与方法：将SRA 01/04细胞分为对照组（CON）、模型组（H2 O2）和治疗组（NR+ H2 O2）。检测细胞超氧化物歧化酶（SOD）、过氧化氢酶（CAT）和总谷胱甘肽（GSH）水平，评价不同浓度H2 O2对SRA 01/04细胞的氧化损伤作用。在用NR和/或H2 O2处理SRA 01/04细胞后，使用3-氨基-3-甲基-N-（2-甲基-N-苯基）-N-（2-甲基）-N采用四甲基偶氮唑蓝（MTT）比色法和Hoechst染色，流式细胞仪检测细胞凋亡，DCFH-DA探针检测活性氧（ROS），MitoTracker染色检测线粒体膜电位（MMP）。此外，进行蛋白质印迹法检测与细胞凋亡和相关信号通路相关的蛋白质的水平。结果：H2 O2通过抑制SOD、CAT活性和降低GSH含量，诱导SRA 01/04细胞氧化损伤。NR处理SRA 01/04细胞显著增加细胞活力，减少细胞凋亡和ROS的产生，而SOD和CAT活性以及总GSH和MMP水平通过NR处理在H2 O2暴露的细胞模型中得到改善。与模型组相比，NR能显著抑制MAPK通路的激活，但促进JAK 2/Stat 3通路的激活。结论：NR可能通过靶向MAPK和JAK 2/Stat 3通路减轻H2 O2诱导的SRA 01/04细胞氧化损伤。NR可能代表一种新的预防或治疗白内障的药物。
ABSTRACT Purpose: To investigate the protective effects of nicotinamide riboside (NR) on oxidative damage in hydrogen peroxide (H2O2)-exposed human lens epithelial cell lines (SRA01/04) and the possible mechanisms underlying its protective effects. Materials and methods: SRA01/04 cells were divided into three groups: the control (CON) group, model (H2O2) group and treatment (NR+H2O2) group. Superoxide dismutase (SOD), catalase (CAT) and total glutathione (GSH) levels were detected to evaluate oxidative damage induced by different concentrations of H2O2 in SRA01/04 cells. After SRA01/04 cells were treated with NR and/or H2O2, cell viability was evaluated using the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay and Hoechst staining, cell apoptosis was analysed using flow cytometry, reactive oxygen species (ROS) were measured with the DCFH-DA probe, and mitochondria were stained with MitoTracker to measure the mitochondrial membrane potential (MMP). In addition, western blotting was performed to detect the levels of proteins associated with apoptosis and related signalling pathways. Results: H2O2 induced oxidative damage in SRA01/04 cells by inhibiting the activity of SOD and CAT and reducing total GSH levels. Treatment of SRA01/04 cells with NR significantly increased cell viability and reduced cell apoptosis and ROS generation, whereas SOD and CAT activities and total GSH and MMP levels were improved by the NR treatment in an H2O2-exposed cell model. Furthermore, NR significantly inhibited the activation of the MAPK pathway but promoted activation of the JAK2/Stat3 pathway compared with the model group. Conclusions: NR may alleviate oxidative damage by targeting the MAPK and JAK2/Stat3 pathways in H2O2-treated SRA01/04 cells. NR may represent anovel drug for preventing or treating cataracts.