Methylcobalamin Protects Melanocytes from H2O2-Induced Oxidative Stress by Activating the Nrf2/HO-1 Pathway

Ran An,1,2 Dong Li,1 Yingying Dong,1 Qiuyun She,1 Ting Zhou,1 Xiaoqi Nie,1 Ronghua Pan,1 Yunhua Deng1 1Department of Dermatology, Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, Hubei, People’s Republic of China; 2Department of Dermatology, Children’s H...

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Autores principales:	An R, Li D, Dong Y, She Q, Zhou T, Nie X, Pan R, Deng Y
Formato:	article
Lenguaje:	EN
Publicado:	Dove Medical Press 2021
Materias:	methylcobalamin vitiligo oxidative stress melanocytes nf-e2-related factor 2 Therapeutics. Pharmacology RM1-950
Acceso en línea:	https://doaj.org/article/431c938c8aef4407859dcf59541e3bc5
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spelling	oai:doaj.org-article:431c938c8aef4407859dcf59541e3bc52021-11-30T18:50:36ZMethylcobalamin Protects Melanocytes from H2O2-Induced Oxidative Stress by Activating the Nrf2/HO-1 Pathway1177-8881https://doaj.org/article/431c938c8aef4407859dcf59541e3bc52021-11-01T00:00:00Zhttps://www.dovepress.com/methylcobalamin-protects-melanocytes-from-h2o2-induced-oxidative-stres-peer-reviewed-fulltext-article-DDDThttps://doaj.org/toc/1177-8881Ran An,1,2 Dong Li,1 Yingying Dong,1 Qiuyun She,1 Ting Zhou,1 Xiaoqi Nie,1 Ronghua Pan,1 Yunhua Deng1 1Department of Dermatology, Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, Hubei, People’s Republic of China; 2Department of Dermatology, Children’s Hospital of Soochow University, Suzhou, Jiangsu, People’s Republic of ChinaCorrespondence: Yunhua DengDepartment of Dermatology, Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology, 1095 Jiefang Ave, Qiaokou, Wuhan, Hubei, People’s Republic of ChinaTel +86 027 83663370Fax +86 027 83663625Email 820331158@qq.comPurpose: 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.Keywords: methylcobalamin, vitiligo, oxidative stress, melanocytes, NF-E2-related factor 2An RLi DDong YShe QZhou TNie XPan RDeng YDove Medical Pressarticlemethylcobalaminvitiligooxidative stressmelanocytesnf-e2-related factor 2Therapeutics. PharmacologyRM1-950ENDrug Design, Development and Therapy, Vol Volume 15, Pp 4837-4848 (2021)
institution	DOAJ
collection	DOAJ
language	EN
topic	methylcobalamin vitiligo oxidative stress melanocytes nf-e2-related factor 2 Therapeutics. Pharmacology RM1-950
spellingShingle	methylcobalamin vitiligo oxidative stress melanocytes nf-e2-related factor 2 Therapeutics. Pharmacology RM1-950 An R Li D Dong Y She Q Zhou T Nie X Pan R Deng Y Methylcobalamin Protects Melanocytes from H2O2-Induced Oxidative Stress by Activating the Nrf2/HO-1 Pathway
description	Ran An,1,2 Dong Li,1 Yingying Dong,1 Qiuyun She,1 Ting Zhou,1 Xiaoqi Nie,1 Ronghua Pan,1 Yunhua Deng1 1Department of Dermatology, Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, Hubei, People’s Republic of China; 2Department of Dermatology, Children’s Hospital of Soochow University, Suzhou, Jiangsu, People’s Republic of ChinaCorrespondence: Yunhua DengDepartment of Dermatology, Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology, 1095 Jiefang Ave, Qiaokou, Wuhan, Hubei, People’s Republic of ChinaTel +86 027 83663370Fax +86 027 83663625Email 820331158@qq.comPurpose: 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.Keywords: methylcobalamin, vitiligo, oxidative stress, melanocytes, NF-E2-related factor 2
format	article
author	An R Li D Dong Y She Q Zhou T Nie X Pan R Deng Y
author_facet	An R Li D Dong Y She Q Zhou T Nie X Pan R Deng Y
author_sort	An R
title	Methylcobalamin Protects Melanocytes from H2O2-Induced Oxidative Stress by Activating the Nrf2/HO-1 Pathway
title_short	Methylcobalamin Protects Melanocytes from H2O2-Induced Oxidative Stress by Activating the Nrf2/HO-1 Pathway
title_full	Methylcobalamin Protects Melanocytes from H2O2-Induced Oxidative Stress by Activating the Nrf2/HO-1 Pathway
title_fullStr	Methylcobalamin Protects Melanocytes from H2O2-Induced Oxidative Stress by Activating the Nrf2/HO-1 Pathway
title_full_unstemmed	Methylcobalamin Protects Melanocytes from H2O2-Induced Oxidative Stress by Activating the Nrf2/HO-1 Pathway
title_sort	methylcobalamin protects melanocytes from h2o2-induced oxidative stress by activating the nrf2/ho-1 pathway
publisher	Dove Medical Press
publishDate	2021
url	https://doaj.org/article/431c938c8aef4407859dcf59541e3bc5
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Methylcobalamin Protects Melanocytes from H2O2-Induced Oxidative Stress by Activating the Nrf2/HO-1 Pathway

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