Protective Effect of Glutathione against Oxidative Stress-induced Cytotoxicity in RAW 264.7 Macrophages through Activating the Nuclear Factor Erythroid 2-Related Factor-2/Heme Oxygenase-1 Pathway
Protective Effect of Glutathione against Oxidative Stress-induced Cytotoxicity in RAW 264.7 Macrophages through Activating the Nuclear Factor Erythroid 2-Related Factor-2/Heme Oxygenase-1 Pathway
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DOI：
10.3390/antiox8040082
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发表时间：
2019-04-01
期刊：
影响因子：
7
通讯作者：
Choi, Yung Hyun
中科院分区：
文献类型：
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作者：
Kwon, Da Hye;Cha, Hee-Jae;Choi, Yung Hyun
活性氧(ROS)是氧化应激的产物，在各种疾病的发生和发展中起重要作用。谷胱甘肽是一种主要的抗氧化剂，可以通过清除ROS来帮助阻止这一过程。本研究旨在评价谷胱甘肽对过氧化氢、过氧化氢诱导的RAW 264.7巨噬细胞ROS损伤和凋亡的保护作用，并探讨核因子红系2相关因子-2(NRF2)/血红素氧合酶-1(HO-1)信号通路的作用。结果表明，H_2O_2处理RAW-264.7细胞后，细胞存活率下降是由于细胞内ROS生成增加而导致DNA损伤和细胞凋亡。而谷胱甘肽能显著逆转过氧化氢诱导的细胞毒性和ROS的产生。此外，H_2O_2诱导的线粒体膜电位的丧失与三磷酸腺苷(ATP)水平的降低有关，而谷胱甘肽的存在也显著减弱了这种变化。这些保护作用伴随着B细胞淋巴瘤-2(Bcl2)/Bcl2相关X蛋白(Bax)的表达增加和caspase-3失活导致的聚(ADP-核糖)聚合酶裂解。此外，谷胱甘肽介导的细胞保护作用与Nrf2的激活和HO-1的表达增加有关；然而，使用HO-1特异性抑制剂锌原卟啉IX抑制HO-1的功能，显著削弱了谷胱甘肽的细胞保护作用。综上所述，这些结果表明，外源性谷胱甘肽能够保护RAW 264.7细胞免受氧化应激诱导的线粒体介导的凋亡以及Nrf2/HO-1信号通路的激活。
Reactive oxygen species (ROS), products of oxidative stress, contribute to the initiation and progression of the pathogenesis of various diseases. Glutathione is a major antioxidant that can help prevent the process through the removal of ROS. The aim of this study was to evaluate the protective effect of glutathione on ROS-mediated DNA damage and apoptosis caused by hydrogen peroxide, H2O2, in RAW 264.7 macrophages and to investigate the role of the nuclear factor erythroid 2-related factor-2 (Nrf2)/heme oxygenase-1 (HO-1) signaling pathway. The results showed that the decrease in the survival rate of RAW 264.7 cells treated with H2O2 was due to the induction of DNA damage and apoptosis accompanied by the increased production of ROS. However, H2O2-induced cytotoxicity and ROS generation were significantly reversed by glutathione. In addition, the H2O2-induced loss of mitochondrial membrane potential was related to a decrease in adenosine triphosphate (ATP) levels, and these changes were also significantly attenuated in the presence of glutathione. These protective actions were accompanied by a increase in the expression rate of B-cell lymphoma-2 (Bcl-2)/Bcl-2-associated X protein (Bax) and poly(ADP-ribose) polymerase cleavage by the inactivation of caspase-3. Moreover, glutathione-mediated cytoprotective properties were associated with an increased activation of Nrf2 and expression of HO-1; however, the inhibition of the HO-1 function using an HO-1 specific inhibitor, zinc protoporphyrin IX, significantly weakened the cytoprotective effects of glutathione. Collectively, the results demonstrate that the exogenous administration of glutathione is able to protect RAW 264.7 cells against oxidative stress-induced mitochondria-mediated apoptosis along with the activity of the Nrf2/HO-1 signaling pathway.