Activation of Skin Cells and Transcription Factors by S*

刺激性皮炎是工作场所受伤的主要原因。它会影响 个人在不同的工作环境中工作，是 严重的发病率和工作时间损失。刺激物的病理生理学 皮炎的定义并不明确。皮肤暴露在刺激性化学物质中 导致炎症的主要特征迅速出现 与细胞因子活化和白细胞浸润有关。刺激性如何 激活炎症尚不清楚。我们认为刺激物可能会激活 先天免疫机制共有的信号通路。以这些为中心 途径是与促炎细胞因子相关的转录因子 引发毛细血管渗漏的基因、细胞黏附分子和蛋白质。一 关键转录因子家族与这些不同的激活相关 基因集是核因子kappaB(NF-kB)转录家族 各种因素。我们的假设是刺激性暴露会导致核因子-kB的激活 在皮肤细胞中。这种激活会触发促炎因子 级联与皮炎的临床表现。为了测试这一点 假设，我们提出了以下具体目标：1)检验 刺激物可激活体外培养的皮肤细胞中的核因子-kB 以内皮细胞为靶细胞。我们将检查刺激性治疗是否 培养的皮肤细胞导致活性的核因子-kB复合体从 检测HDMEC、HK或HDMEC的刺激性处理 HDF诱导IKBA的磷酸化和泛素化，必要的步骤 蛋白小体介导的降解；检查移位的核因子-kB复合体 能够与细胞因子和细胞的相关反应元件结合 黏附分子基因启动子。2)检查刺激物是否可以激活 用上皮细胞和内皮细胞体外培养皮肤细胞中的特异性基因 以细胞为目标。我们将研究刺激物是否会诱导或上调 黏附分子ICAM-1和E-选择素的表达与促炎作用 细胞因子IL-8和VEGF-C；检测刺激物是否增加稳态mRNA ICAM-1、E-选择素、IL-8或VEGF-C的表达；检查是否有刺激性 培养的皮肤细胞处理导致黏附转录增加 依赖于核因子-kB反应元件的分子或细胞因子基因。这些 研究将提供一个框架，以确定刺激物 导致皮肤发炎，并提供合理和机械的靶点 用于预防和治疗。

Irritant dermatitis is a major cause of workplace injury. It affects individuals in a variety of work environments and is a source of significant morbidity and lost work time. The pathophysiology of irritant dermatitis is only poorly defined. Exposure of the skin to irritant chemicals results in the rapid appearance of the cardinal features of inflammation associated with cytokine activation and leukocyte infiltration. How irritants activate inflammation is not known. We believe that irritants may activate signaling pathways common to innate immune mechanisms. Centered in these pathways are transcription factors that are linked to pro-inflammatory cytokine genes, cell adhesion molecules, and proteins that trigger capillary leak. One key family of transcription factors linked to the activation of these diverse gene sets is the nuclear factor kappa B (NF-kB) family of transcription factors. Our hypothesis is that irritant exposure results in NF-kB activation in skin cells. This activation results in the triggering of a pro-inflammatory cascade and the clinical appearance of dermatitis. In order to test this hypothesis, we propose the following specific aims: 1) To examine whether irritants can activate NF-kB in cultured skin cells in vitro using epithelial and endothelial cells as targets. We will examine whether irritant treatment of cultured skin cells results in translocation of active NF-kB complexes from the cytoplasm to the nucleus; examine whether irritant treatment of HDMEC, HK, or HDF induces the phosphorylation and ubiquitination of IkBa, necessary steps for proteosome mediated degradation; examine whether translocated NF-kB complexes are capable of binding to relevant response elements of cytokine and cell adhesion molecule gene protomers. 2) To examine whether irritants can activate specific genes in cultured skin cells in vitro using epithelial and endothelial cells as targets. We will examine whether irritants induce or upregulate the expression of adhesion molecules ICAM-1 and E-selectin, and proinflammatory cytokines IL-8 and VEGF-C; examine whether irritants increase steady state mRNA expression of ICAM-1, E-selectin, IL-8, or VEGF-C; examine whether irritant treatment of cultured skin cells results in increase transcription of adhesion molecule or cytokine genes dependent upon NF-kB responsive elements. These studies will provide a framework to define the mechanisms by which irritants cause inflammation in the skin and provide a rational and mechanistic targets for prevention and treatment.