Li Ziyu, Yu Yongkai, Cao Xuechen, Wang Yidan, Lu Jiawei, Feng Yifei, Jiang Yali, Lu Yan
Department of Dermatology, the First Affiliated Hospital of Nanjing Medical University, Nanjing, 210029, China.
The Friendship Hospital of Ili Kazakh Autonomous Prefecture, Xinjiang Uygur Autonomous Region, Xinjiang, 835000, China.
Mol Biol Rep. 2025 Feb 12;52(1):223. doi: 10.1007/s11033-025-10329-1.
Vitiligo is a common depigmentation disorder. Oxidative stress in melanocytes is thought to be the primary cause of vitiligo. Imbalances in cellular calcium ion (Ca) levels may be associated with the onset and progression of various diseases through a process that has been linked to oxidative stress. The purpose of this study was to investigate the regulatory mechanism by which Ca levels change in normal human melanocytes (NHMs) under oxidative stress, thereby providing new insights and potential clinical therapeutic targets for the pathogenesis and treatment of vitiligo.
Single-cell RNA sequencing data from vitiligo patients were analyzed using bioinformatics techniques. NHMs were treated with hydrogen peroxide (HO), store-operated Ca entry (SOCE) blocker BTP2, and SOCE agonist cyclopiazonic acid. Flow cytometry was used to detect Ca levels, apoptosis rates, intra-mitochondrial reactive oxygen species (ROS) levels, and mitochondrial membrane potential (MMP) damage. The expression levels of target proteins were detected using immunofluorescence, quantitative real-time PCR, and western blotting. We found that HO-induced oxidative stress resulted in significantly increased intracellular Ca levels, upregulation of stromal interaction molecule 1 (STIM1) and calcium release-activated calcium channel protein (ORAI1), and mitochondrial dysfunction. Inhibition of SOCE and small interfering RNA-mediated silencing of STIM1/ORAI1 expression lowered mitochondrial levels of ROS and oxidative stress-induced intracellular Ca overload and restored MMP, ultimately terminating oxidative stress-induced apoptosis.
Oxidative stress upregulates STIM1/ORAI1 expression, leading to melanocyte apoptosis via increased Ca influx, whereas inhibition of SOCE protects melanocytes against oxidative stress-induced damage.
白癜风是一种常见的色素脱失性疾病。黑素细胞中的氧化应激被认为是白癜风的主要病因。细胞钙离子（Ca）水平失衡可能通过与氧化应激相关的过程，与各种疾病的发生和发展有关。本研究的目的是探讨氧化应激下正常人黑素细胞（NHM）中Ca水平变化的调控机制，从而为白癜风的发病机制和治疗提供新的见解和潜在的临床治疗靶点。
使用生物信息学技术分析白癜风患者的单细胞RNA测序数据。用过氧化氢（HO）、储存式钙内流（SOCE）阻滞剂BTP2和SOCE激动剂环匹阿尼酸处理NHM。采用流式细胞术检测Ca水平、凋亡率、线粒体内活性氧（ROS）水平和线粒体膜电位（MMP）损伤。使用免疫荧光、定量实时PCR和蛋白质印迹法检测靶蛋白的表达水平。我们发现HO诱导的氧化应激导致细胞内Ca水平显著升高、基质相互作用分子1（STIM1）和钙释放激活钙通道蛋白（ORAI1）上调以及线粒体功能障碍。抑制SOCE以及小干扰RNA介导的STIM1/ORAI1表达沉默降低了线粒体ROS水平和氧化应激诱导的细胞内Ca超载，并恢复了MMP，最终终止了氧化应激诱导的凋亡。
氧化应激上调STIM1/ORAI1表达，通过增加Ca内流导致黑素细胞凋亡，而抑制SOCE可保护黑素细胞免受氧化应激诱导的损伤。
