Berberine protects immortalized line of human melanocytes from H2O2-induced oxidative stress via activation of Nrf2 and Mitf signaling pathway
Berberine protects immortalized line of human melanocytes from H2O2-induced oxidative stress via activation of Nrf2 and Mitf signaling pathway
复制标题
小檗碱通过激活 Nrf2 和 Mitf 信号通路保护永生化的人类黑素细胞系免受 H2O2 诱导的氧化应激
DOI：
10.1016/j.jdermsci.2019.03.007
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发表时间：
2019-04-01
影响因子：
4.6
通讯作者：
Jian, Zhe
中科院分区：
文献类型：
--
作者：
Jiang, Wei;Li, Shuli;Jian, Zhe
背景：氧化应激在白癜风的发病机制中起重要作用。清除过氧化氢(H_2O_2)已被证实对白癜风患者有益。黄连素(BBR)是一种天然的异喹啉生物碱，具有抗氧化活性，但BBR是否能保护人黑素细胞免受氧化损伤尚不清楚。目的：研究BBR对永生化人黑素细胞株PIG1氧化应激的保护作用。方法：体外培养的PIG1细胞经不同浓度的BBR处理1h后，再暴露于1.0 mM H_2O_2中24 h，通过流式细胞仪检测细胞凋亡率、细胞内活性氧(ROS)水平。结果：经BBR处理后，细胞活力增加，细胞内ROS生成增加，细胞凋亡率降低，细胞凋亡率降低。此外，我们还发现，BBR可以显著诱导Nrf2核转位，增加总Nrf2水平，增强ARE活性。此外，Nrf2-siRNA的转染可以消除BBR对黑素细胞氧化损伤的保护作用。最后，我们验证了BBR通过对MITF及其靶蛋白的调节来促进黑素细胞的功能。结论：BBR可能通过其抗氧化活性保护黑素细胞免受氧化应激的伤害。此外，我们还发现BBR抑制了过氧化氢诱导的核因子kappaB的激活。因此，BBR作为治疗白癜风的潜在药物值得研究。(C)2019年，爱思唯尔公司代表日本皮肤病研究学会出版。
Background: Oxidative stress plays important roles in the pathogenesis of vitiligo. The removal of hydrogen peroxided (H2O2) has been established to be beneficial to vitiligo patients. Berberine (BBR), a natural isoquinoline alkaloid, has antioxidant activity, however, whether BBR can defend human melanocytes against oxidative injury remains to be elucidated.Objective: In the present study, we investigated the potential protective effect of BBR against oxidative stress on an immortalized normal human melanocyte cell line PIG1.Methods: Generally, PIG1 cells were pretreated with various concentrations of BBR for 1 h followed by exposure to 1.0 mM H2O2 for 24 h. Cell apoptosis, intracellular reactive oxygen species (ROS) levels were assessed through flow cytometry. Cell apoptosis, melanogenesis and the activation of Nrf2-ARE and Mitf signaling pathway were assayed.Results: Our results showed that cell viability rose and intracellular ROS generation, cell apoptosis of melanocytes decreased significantly in response to H2O2 through pretreatment with BBR. Furthermore, We found that BBR can dramatically induce Nrf2 nuclear translocation, increase total Nrf2 levels and enhance ARE activity. Besides, Nrf2-siRNA transfection can abrogate the protection of BBR in melanocytes against oxidative injury. At last, we verified that BBR could facilitate melanogenesis function via modulation of Mitf and its target proteins.Conclusion: The results above suggest that BBR can protect melanocytes against oxidative stress via its anti-oxidative activity. Also, we found H2O2-induced activation of NF kappa B was inhibited by BBR. Therefore, it is worthy of investigation BBR as a potential drug for treatment of vitiligo. (C) 2019 Published by Elsevier B.V. on behalf of Japanese Society for Investigative Dermatology.