Methylcobalamin Protects Melanocytes from H(2)O(2)-Induced Oxidative Stress by Activating the Nrf2/HO-1 Pathway.
Methylcobalamin Protects Melanocytes from H(2)O(2)-Induced Oxidative Stress by Activating the Nrf2/HO-1 Pathway.
复制标题
甲钴胺通过激活 Nrf2/HO-1 通路保护黑素细胞免受 H2O2 诱导的氧化应激
DOI：
10.2147/dddt.s336066
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发表时间：
2021
期刊：
影响因子：
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通讯作者：
Deng Y
中科院分区：
文献类型：
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作者：
An R;Li D;Dong Y;She Q;Zhou T;Nie X;Pan R;Deng Y
目的氧化应激被认为是白癜风发病的主要决定因素。甲钴胺（MeCbl）是维生素B12的活化形式，可调节炎症因子，对抗氧化应激，并减少许多疾病模型中的细胞凋亡。然而，MeCbl再色素沉着对白癜风的具体机制尚不清楚。在这项研究中，我们探讨了MeCbl对黑素细胞过氧化氢（H2 O2）诱导的氧化应激的影响。方法采用人黑素细胞株PIG 1建立氧化应激模型。我们使用细胞计数试剂盒-8（CCK-8）检测药物的细胞毒性，并使用NaOH法测量细胞的黑色素含量。通过流式细胞术和抗氧化酶检测试剂盒评估细胞内氧化损伤。此外，我们通过流式细胞术和Western印迹法评估了细胞凋亡的存在。我们探讨了MeCbl在黑素细胞氧化应激过程中的潜在机制，通过实时定量聚合酶链反应（RT-qPCR），蛋白质印迹和激光扫描共聚焦免疫荧光显微镜的基础上分析实验结果。最后，我们在应用抑制剂阻断Nrf 2通路后重复实验。结果MeCbl处理可提高细胞活力，增加黑素含量，减少细胞内活性氧（ROS）的积累，提高抗氧化酶超氧化物歧化酶（SOD）和过氧化氢酶（CAT）的活性，减少黑素细胞凋亡，上调Nrf 2/HO-1通路的表达。抑制Nrf 2/HO-1通路后，MeCbl的保护作用明显减弱。结论MeCbl可通过激活Nrf 2/HO-1通路减轻H2 O2诱导的黑素细胞氧化应激，提示MeCbl可能是治疗白癜风的有效药物。
Purpose Oxidative stress is considered a major determinant in the pathogenesis of vitiligo. Methylcobalamin (MeCbl) is an activated form of vitamin B12 that regulates inflammatory factors, counters oxidative stress, and reduces apoptosis in many disease models. However, the specific mechanism of MeCbl repigmentation against vitiligo is unknown. In this study, we explored the effect of MeCbl on melanocytes following hydrogen peroxide (H2O2)-induced oxidative stress. Methods We established an oxidative stress model using the immortalized human normal melanocyte cell line PIG1. We used a Cell Counting Kit-8 (CCK-8) to detect drug cytotoxicity, and we measured the melanin content of cells using the NaOH method. Intracellular oxidative damage was assessed by flow cytometry and antioxidant enzyme detection kits. In addition, we assessed the presence of apoptosis by flow cytometry and Western blots. We explored the underlying mechanisms of MeCbl during oxidative stress in melanocytes by analyzing the results of experiments based on real-time quantitative polymerase chain reaction (RT-qPCR), Western blotting, and laser scanning confocal immunofluorescence microscopy. Finally, we repeated the experiments after applying an inhibitor to block the Nrf2 pathway. Results We found that MeCbl treatment enhanced cell viability, increased melanin content, reduced intracellular reactive oxygen species (ROS) accumulation, increased the activities of antioxidant enzyme superoxide dismutase (SOD) and catalase (CAT), reduced melanocyte apoptosis, and up-regulated the expression of the Nrf2/HO-1 pathway. Moreover, the protective effects of MeCbl were significantly weakened after inhibiting the Nrf2/HO-1 pathway. Conclusion Our results indicate that MeCbl attenuated the H2O2-induced oxidative stress in melanocytes by activating the Nrf2/HO-1 pathway, this suggests that MeCbl may be an effective treatment against vitiligo.