Abstract：
Vitiligo is featured by manifestation of white maculae and primarily results from oxidative stress. Sphingosine kinase-1 (SPHK1) participates in oxidative stress. This paper was devised to explore the role of SPHK1 in vitiligo and to disclose the mechanism. PIG1 cell viability was appraised utilizing cell counting kit-8 assay while Western blot detected SPHK1 and four and a half LIM domains 2 (FHL2). The transduction efficacy of small interfering RNA (siRNA)-SPHK1, siRNA-FHL2 and pcDNA3.1 plasmid overexpressing FHL2 (Ov-FHL2) was checked using Western blot. Flow cytometry detected cell apoptotisis. Western blot detected mitochondrial cytochrome c (Mit-Cyt-c) and cytosolic cytochrome c (Cyto-Cyt-c). Dichloro-dihydro-fluorescein diacetate (DCFH-DA) detected reactive oxygen species (ROS) activity while oxidative stress markers were evaluated using corresponding assay kits. SPHK1 expression was discovered to be increased in hydrogen peroxide (H2O2)-challenged PIG1 cells and SPHK1 interference alleviated H2O2-challenged viability damage, apoptosis, oxidative stress and FHL2 expression in PIG1 cells. FHL2 depletion could suppress viability damage, apoptosis and oxidative stress in H2O2-challenged PIG1 cells. Rescue experiments demonstrated that the suppressive impacts of SPHK1 deficiency on PIG1 cell viability, apoptosis and oxidative stress induced by H2O2 were offset by FHL2 overexpression. Collectively, SPHK1 knockdown protected against vitiligo via the regulation of FHL2.
摘要：
白癜风以白色黄斑表现为特征，主要由氧化应激引起。鞘氨醇激酶-1(SPHK1)参与氧化应激。本文旨在探讨SPHK1在白癜风中的作用并揭示其机制。使用细胞计数试剂盒-8测定法评估PIG1细胞活力，而Western印迹检测SPHK1和四个半LIM结构域2（FHL2）。使用蛋白质印迹检查过表达FHL2(Ov-FHL2)的小干扰RNA(siRNA)-SPHK1、siRNA-FHL2和pcDNA3.1质粒的转导功效。流式细胞术检测细胞凋亡。Westernblot检测到线粒体细胞色素c(Mit-Cyt-c)和胞浆细胞色素c(Cyto-Cyt-c)。二氯-二氢-荧光素二乙酸酯（DCFH-DA）检测到活性氧（ROS）活性，而使用相应的测定试剂盒评估氧化应激标志物。发现SPHK1表达在过氧化氢（H2O2）攻击的PIG1细胞中增加，SPHK1干扰减轻了H2O2攻击的生存力损伤，凋亡，氧化应激和FHL2在PIG1细胞中的表达。FHL2消耗可以抑制生存力损伤，H2O2攻击的PIG1细胞的凋亡和氧化应激。挽救实验表明，SPHK1缺乏对PIG1细胞活力的抑制作用，FHL2过表达抵消了H2O2诱导的细胞凋亡和氧化应激。总的来说，SPHK1敲除通过调节FHL2保护白癜风。
