Abstract Vitiligo is caused by the disappearance of melanocyte function in the skin, but the mechanism has not been fully elucidated. This study aimed to investigate the expression and role of miR-217 in dysfunctional human primary melanocytes caused by oxidative stress, so as to explore its potential role in vitiligo. Hydrogen peroxide (H2O2) was used to induce the oxidative damage of melanocytes. The levels of sirtuin 1 (SIRT1) and miR-217 were measured by quantitative reverse transcription-polymerase chain reaction (qRT-PCR) and or Western blot assay. TargetScan and dual luciferase reporter gene assay were used to confirm the relationship between SIRT1 and miR-217. Cell viability and cell apoptosis were analysed by 3-(4,5-dimethyl-2-thiazolyl)−2,5-diphenyl-2-H-tetrazolium bromide (MTT) assay and flow cytometry assay, respectively. Reactive oxygen species (ROS) production, superoxide dismutase (SOD) and catalase (CAT) activities were determined using specific assay kits. SIRT1 was down-regulated, while miR-217 was up-regulated in H2O2-induced human primary melanocytes. SIRT1 was a target gene of miR-217. Inhibition of miR-217 increased cell viability and inhibited cell apoptosis in H2O2-treated melanocytes. Besides, inhibition of miR-217 enhanced the antioxidant activity of SOD and CAT and reduced the accumulation of intracellular ROS. Notably, all these effects were reversed by SIRT1-siRNA. The data indicated that SIRT1 was down-regulated, while miR-217 was up-regulated in dysfunctional human primary melanocytes caused by oxidative stress, and miR-217 inhibition relieved oxidative stress-induced melanocyte damage via targeting SIRT1, indicating that miR-217 might a potential target for vitiligo treatment.

中文翻译：

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MiR-217抑制通过靶向sirtuin 1减轻氧化应激诱导的黑素细胞损伤

摘要 白癜风是皮肤中黑色素细胞功能消失所致，但其机制尚未完全阐明。本研究旨在探讨miR-217在氧化应激引起的人原代黑素细胞功能障碍中的表达及作用，以探讨其在白斑病中的潜在作用。过氧化氢 (H2O2) 用于诱导黑素细胞的氧化损伤。Sirtuin 1 (SIRT1) 和 miR-217 的水平通过定量逆转录聚合酶链反应 (qRT-PCR) 和/或蛋白质印迹测定法测量。TargetScan 和双荧光素酶报告基因检测用于确认 SIRT1 和 miR-217 之间的关系。细胞活力和细胞凋亡分别通过 3-(4,5-二甲基-2-噻唑基)-2,5-二苯基-2-H-溴化四唑 (MTT) 测定和流式细胞术测定进行分析。使用特定检测试剂盒测定活性氧 (ROS) 产生、超氧化物歧化酶 (SOD) 和过氧化氢酶 (CAT) 活性。在 H2O2 诱导的人类原代黑素细胞中，SIRT1 下调，而 miR-217 上调。SIRT1 是 miR-217 的靶基因。在 H2O2 处理的黑素细胞中，miR-217 的抑制增加了细胞活力并抑制了细胞凋亡。此外，抑制 miR-217 可增强 SOD 和 CAT 的抗氧化活性并减少细胞内 ROS 的积累。值得注意的是，所有这些影响都被 SIRT1-siRNA 逆转。数据表明，在氧化应激引起的功能失调的人类原代黑素细胞中，SIRT1 下调，而 miR-217 上调，抑制 miR-217 通过靶向 SIRT1 缓解氧化应激诱导的黑素细胞损伤，