Koren Carmi I, Haj R, Yehuda H, Tamir S, Reznick A Z
Department of Anatomy and Cell Biology, Rappaport Faculty of Medicine, Technion - Israel Institute of Technology, 9649, Haifa, Israel,
Adv Exp Med Biol. 2015;849:1-10. doi: 10.1007/5584_2014_98.
Oxidative stress (OS) is common in inflammatory conditions and may be important in atopic dermatitis (AD) etiology. The aim of this project was to study the involvement of oxidation in FSL-1 (deacylated lipoprotein)-triggered signaling pathways leading to AD-typical cytokine expression in HaCaT keratinocytes. HaCaT keratinocytes, pretreated with the inhibitor to OS N-acetylcysteine (NAC), were exposed to FSL-1, a stimulator of AD-related cytokines. Cytokines expression was studied by real time polymerase chain reaction (PCR); nuclear factor-kappa B (NF-κB) and p38 mitogen activated protein kinase (MAPK) activities were studied by western blotting; and the oxidative state of cells was determined by the dichlorofluorescein (DCF) assay. We found that endogenous OS in keratinocytes appeared 4 h after FSL-1 administration. OS activated NF-κB, but not p38 MAPK, and the inhibition of OS reduced FSL-1 induced interleukin (IL) 33, thymic stromal lymphopoietin (TSLP) and TNFα mRNA expression. We conclude that FSL-1 triggers an OS reaction in HaCaT keratinocytes, which is probably a secondary event affecting the expression of specific AD typical cytokines, possibly through the NF-κB pathways. This role of OS in the inflammatory response in AD is worth further investigating.
氧化应激（OS）在炎症性疾病中很常见，可能在特应性皮炎（AD）的病因学中起重要作用。本项目的目的是研究氧化作用在FSL-1（去酰基脂蛋白）触发的信号通路中的参与情况，该信号通路导致HaCaT角质形成细胞中出现AD典型的细胞因子表达。用OS抑制剂N-乙酰半胱氨酸（NAC）预处理的HaCaT角质形成细胞暴露于FSL-1，FSL-1是一种AD相关细胞因子的刺激物。通过实时聚合酶链反应（PCR）研究细胞因子表达；通过蛋白质印迹法研究核因子-κB（NF-κB）和p38丝裂原活化蛋白激酶（MAPK）的活性；并通过二氯荧光素（DCF）测定法确定细胞的氧化状态。我们发现，在给予FSL-1后4小时，角质形成细胞中的内源性OS出现。OS激活了NF-κB，但未激活p38 MAPK，并且抑制OS可降低FSL-1诱导的白细胞介素（IL）33、胸腺基质淋巴细胞生成素（TSLP）和肿瘤坏死因子α（TNFα）mRNA表达。我们得出结论，FSL-1在HaCaT角质形成细胞中触发了OS反应，这可能是一个影响特定AD典型细胞因子表达的继发事件，可能是通过NF-κB途径。OS在AD炎症反应中的这一作用值得进一步研究。
