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建立使用细胞因子组合刺激的 HaCaT 细胞的银屑病样皮肤体外转录谱。

Establishing Transcription Profile of Psoriasiform Cutaneous In Vitro using HaCaT Cells Stimulated with Combination of Cytokines.

机构信息

State Key Laboratory of Biotherapy and Cancer Center, West China Hospital, West China Medical School, Sichuan University and Collaborative Innovation Center for Biotherapy.

Department of Cardiovascular Medicine, West China Hospital, Sichuan University.

出版信息

J Vis Exp. 2021 Mar 15(169). doi: 10.3791/61537.

DOI:10.3791/61537
PMID:33779603
Abstract

Psoriasis is a common chronic inflammatory skin disease mediated by innate and adaptive immune systems, characterized by abnormal proliferation and differentiation of epidermal keratinocytes and infiltration of inflammatory cells. Skin-specific keratinocytes are key participants in innate immunity, responding to immune cells and environmental stimulation, thereby serving an important role in the immunopathogenesis of psoriasis. Here, we present a method for inducing psoriasiform keratinocytes inflammation at transcription level with HaCaT cell line using five proinflammatory cytokines combination (M5 combination), including IL-17A, IL-22, IL-1α, TNF-α, and oncostatin M. Results demonstrate that M5 combination induced HaCaT cells showed increased levels of antimicrobial peptides (BD2, S100A7, S100A8, and S100A9), chemokines, and cytokines (CXCL1, CXCL2, CXCL8, CCL20, IL-1β, IL-6 and, IL-18). The mRNA levels of keratinocytes differentiation markers (Keratin1, Keratin10, Filaggrin, and Loricrin) were down regulated, which was consistent with the transcriptome data derived from psoriasis-like keratinocytes. The method described here, therefore, establishes an in vitro psoriasiform cutaneous inflammation at transcription level and contributes to the research for molecular pathogenesis of psoriasis.

摘要

银屑病是一种由先天和适应性免疫系统介导的常见慢性炎症性皮肤病,其特征是表皮角质形成细胞的异常增殖和分化以及炎症细胞的浸润。皮肤特异性角质形成细胞是先天免疫的关键参与者,对免疫细胞和环境刺激作出反应,从而在银屑病的免疫发病机制中发挥重要作用。在这里,我们提出了一种使用五种促炎细胞因子组合(M5 组合)在 HaCaT 细胞系上诱导银屑病角质形成细胞炎症的转录水平的方法,该组合包括白细胞介素 17A(IL-17A)、白细胞介素 22(IL-22)、白细胞介素 1α(IL-1α)、肿瘤坏死因子-α(TNF-α)和癌蛋白 M(Oncostatin M)。结果表明,M5 组合诱导的 HaCaT 细胞显示抗菌肽(BD2、S100A7、S100A8 和 S100A9)、趋化因子和细胞因子(CXCL1、CXCL2、CXCL8、CCL20、白细胞介素 1β(IL-1β)、白细胞介素 6(IL-6)和白细胞介素 18(IL-18)水平升高。角质形成细胞分化标志物(角蛋白 1、角蛋白 10、丝聚合蛋白和兜甲蛋白)的 mRNA 水平下调,与银屑病样角质形成细胞的转录组数据一致。因此,这里描述的方法在转录水平上建立了一种体外银屑病样皮肤炎症,有助于银屑病发病机制的分子研究。

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