Reactive oxygen species (ROS) is mainly produced as a by-product from electron transport chain (ETC) of mitochondria and effectively eliminated by cellular antioxidants. However, 2-chloroethyl ethyl sulfide (CEES) exposure to keratinocytes declined antioxidant capacity and increased accumulation of ROS triggered alteration of mitochondrial activity and apoptosis is lacking. Our findings demonstrated that the electron leakage from the impaired ETC, leading to the accumulation of ROS was gradually elevating with increasing concentration of CEES exposure, which decline the activity of superoxide dismutase (SOD), manganese SOD (MnSOD) and copper-zinc SOD (Cu-ZnSOD) in keratinocytes. Further, excess accumulation of ROS, decreased the mitochondrial membrane potential (ΔΨm) and increased the mitochondrial mass with increasing dose of CEES. CEES exposure provoked the decrease in expression of transcription factor A mitochondrial (TFAM), augmented mitochondrial DNA (mtDNA) damage and altered the mtDNA-encoded oxidative phosphorylation (OXPHOS) subunits. Moreover, fragmented mtDNA translocated into cytosol, where it activated cGAS-STING and interferon regulatory factor3 (IRF3), coinciding with the increased expression of inflammatory mediators and alteration of cell-to-cell communication markers. Pre-treatment of N-acetyl-l-cysteine (NAC) or L-Nω-nitroarginine methyl ester (NAME), hydralazine hydrochloride (Hyd·HCl) or ERK1/2 or phosphoinositide3-kinase (PI3–K)/Akt inhibitors in keratinocyte cells significantly restored the CEES effect. Our findings suggest that CEES-induced mitochondrial ROS production and accumulation leads to mitochondrial dysfunction and inflammatory response in keratinocytes. However, treatment of antioxidants or ERK1/2 or PI3–K/Akt inhibitors is a novel therapeutic option for the keratinocytes complication.

活性氧（ROS）主要是由线粒体的电子传输链（ETC）产生的副产物，并被细胞抗氧化剂有效消除。然而，2-氯乙基乙基硫化物（CEES）暴露于角质形成细胞会降低抗氧化能力，并且ROS积累增加会触发线粒体活性的改变，并且缺乏细胞凋亡。我们的发现表明，随着CEES暴露浓度的增加，受损的ETC引起​​的电子泄漏导致ROS的积累逐渐增加，从而降低了超氧化物歧化酶（SOD），锰SOD（MnSOD）和铜锌SOD的活性（角质形成细胞中的Cu-ZnSOD）。此外，随着CEES剂量的增加，ROS的过量积聚降低了线粒体膜电位（ΔΨm）并增加了线粒体质量。CEES暴露引起转录因子A线粒体（TFAM）的表达减少，线粒体DNA（mtDNA）损伤增加并改变了mtDNA编码的氧化磷酸化（OXPHOS）亚基。此外，片段化的mtDNA易位到细胞质中，在那里它激活cGAS-STING和干扰素调节因子3（IRF3），与炎症介质表达的增加和细胞间通信标记的改变相吻合。N-乙酰基的预处理 与炎性介质表达的增加和细胞间通讯标记的改变相吻合。N-乙酰基的预处理 与炎性介质表达的增加和细胞间通讯标记的改变相吻合。N-乙酰基的预处理角质形成细胞中的l-半胱氨酸（NAC）或L-Nω-硝基精氨酸甲酯（NAME），盐酸肼屈嗪（Hyd·HCl）或ERK1 / 2或磷酸肌醇3激酶（PI3-K）/ Akt抑制剂可显着恢复CEES效应。我们的发现表明，CEES诱导的线粒体ROS的产生和积累导致角质形成细胞中的线粒体功能障碍和炎症反应。然而，抗氧化剂或ERK1 / 2或PI3–K / Akt抑制剂的治疗是角质形成细胞并发症的一种新型治疗选择。