Pastore S, Giustizieri M L, Mascia F, Giannetti A, Kaushansky K, Girolomoni G
Laboratory of Immunology, Istituto Dermopatico dell'Immacolata, IRCCS, Rome, Italy.
J Invest Dermatol. 2000 Dec;115(6):1134-43. doi: 10.1046/j.1523-1747.2000.00149.x.
Keratinocytes of patients with atopic dermatitis produce high amounts of granulocyte/macrophage colony-stimulating factor, a factor essential for dendritic cell function and thus for the development of skin immune responses. In contrast to keratinocytes cultured from nonatopic, healthy individuals, granulocyte/macrophage colony-stimulating factor mRNA could be detected in unstimulated cultures of atopic dermatitis keratinocytes, and phorbol myristate acetate induced much greater granulocyte/macrophage colony-stimulating factor mRNA levels in these cells, although the decay kinetics were not altered. Using reporter gene (chloramphenicol acetyl transferase) analysis, a minimal granulocyte/macrophage colony-stimulating factor promoter was shown to confer constitutive and phorbol-myristate-acetate-induced regulation of transcriptional activity in keratinocytes, and significantly higher levels of chloramphenicol acetyl transferase activity were measured in lysates of unstimulated and phorbol-myristate-acetate-treated atopic dermatitis keratinocytes than in control keratinocyte cultures. Electrophoretic mobility shift assays showed that low levels of NF-kappa B binding activity could be induced by phorbol myristate acetate in both normal and atopic dermatitis keratinocytes. By contrast, activator protein 1 complexes were efficiently induced, and they were invariably present at higher levels in nuclear lysates of atopic dermatitis keratinocytes. Atopic dermatitis keratinocyte nuclear lysates had higher constitutive levels of c-Jun, and phorbol myristate acetate promoted an earlier and stronger expression of c-Jun, JunB, and of the phosphorylated forms of c-Fos. A dysregulated activation of activator protein 1 may be implicated in the molecular mechanisms leading to increased granulocyte/macrophage colony-stimulating factor expression in atopic dermatitis keratinocytes. J Invest Dermatol 115:1134-1143 2000
特应性皮炎患者的角质形成细胞会产生大量粒细胞/巨噬细胞集落刺激因子，该因子对树突状细胞功能至关重要，因而对皮肤免疫反应的发展也至关重要。与从非特应性健康个体培养的角质形成细胞不同，在未受刺激的特应性皮炎角质形成细胞培养物中可检测到粒细胞/巨噬细胞集落刺激因子mRNA，佛波酯肉豆蔻酸酯乙酸盐可诱导这些细胞中粒细胞/巨噬细胞集落刺激因子mRNA水平大幅升高，尽管其衰减动力学未改变。通过报告基因（氯霉素乙酰转移酶）分析表明，最小的粒细胞/巨噬细胞集落刺激因子启动子可赋予角质形成细胞转录活性的组成型和佛波酯肉豆蔻酸酯乙酸盐诱导的调节，并且在未受刺激的和经佛波酯肉豆蔻酸酯乙酸盐处理的特应性皮炎角质形成细胞裂解物中测得的氯霉素乙酰转移酶活性水平明显高于对照角质形成细胞培养物。电泳迁移率变动分析表明，佛波酯肉豆蔻酸酯乙酸盐可在正常和特应性皮炎角质形成细胞中诱导低水平的核因子κB结合活性。相比之下，激活蛋白1复合物被有效诱导，并且它们在特应性皮炎角质形成细胞核裂解物中的水平始终更高。特应性皮炎角质形成细胞核裂解物中c-Jun的组成型水平较高，佛波酯肉豆蔻酸酯乙酸盐可促进c-Jun、JunB以及c-Fos磷酸化形式的更早、更强表达。激活蛋白1的失调激活可能与导致特应性皮炎角质形成细胞中粒细胞/巨噬细胞集落刺激因子表达增加的分子机制有关。《皮肤病学研究杂志》115:1134 - 1143 2000
Zotero 插件