ROS stress responses of primary human epidermal keratinocytes
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ABSTRACT:
PROVIDER: PRJNA414068 | ENA |
REPOSITORIES: ENA
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Project description:Analysis of cultured epidermal keratinocytes treated with interleukin-4 (IL-4) and interleukin-13 (IL-13). IL-4 and IL-13 are up-regulated in atopic dermatitis. Results provide insight into the role of IL-4 and IL-13 cytokines in the pathogenesis of atopic dermatitis. Analysis of epidermal keratinocytes transfected with dual oxidase 1 (DUOX1) siRNA knockdown before treatment with IL-4 and IL-13. DUOX1 is one of the NOX family members of NADPH oxidases whose primary function is ROS generation. Results provide insight into the role of the incraesed expression of DUOX1 in IL-4/IL-13-treated NHEK for IL4/IL13 signaling. IL-4 and IL-13 induced gene expression in human epidermal keratinocytes (NHEK) was measured at 48 hours. Gene expression in NHEK tranfected with 10 nM DUOX1 siRNA followed by treatment with 100 ng/ml IL-4 and 100 ng/ml IL-13 was measured at 48 hours. Three independent experiments were performed using different strains for each experiment.
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2010-10-31 | GSE20706 | GEO
Project description:The study aimed to interrogate whether the 1,4-naphtoquinone derivative lawsone activates the aryl hydrocarbon receptor (AhR) downstream transcriptional program in keratinocytes and is capable of modulating skin homeostasis. The study has been performed on human primary epidermal keratinocytes (HEKs), which represent the first line of defense against exogenous molecules. HEK cells were treated with the vehicle control DMSO (control), stimulated with lawsone or with the TLR2 ligand Pam2CSK4. Transcriptional profiles of HEK cells were studied to investigate the regulation of AhR -related genes, Nrf2-related genes and epidermal differentiation and keratin genes. We demonstrated that lawsone was sensed by keratinocytes and activated AhR. In particular, lawsone efficiently activated the transcriptional program of AhR and promoted keratinocyte differentiation.
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2014-10-02 | E-GEOD-53751 | biostudies-arrayexpress
Project description:Glucocorticoids (GCs) have a long history of use as therapeutic agents for numerous skin diseases. Surprisingly, their specific molecular effects are largely unknown. To characterize GC action in epidermis, we compared the transcriptional profiles of primary human keratinocytes untreated and treated with dexamethasone (DEX) for 1, 4, 24, 48 and 72 hours using large-scale microarray analyses. The majority of genes were found regulated only after 24 hours and remained regulated throughout the treatment. In addition to expected anti-inflammatory genes, we found that GCs regulate cell fate, tissue remodeling, cell motility, differentiation and metabolism. GCs not only effectively block signaling by TNF-alpha and IL-1 but also by IFN-gamma, which was not previously known. Specifically, GCs suppress the expression of essentially all IFN-gamma-regulated genes, including IFN-gamma receptor and STAT-1. GCs also block STAT-1 activation and nuclear translocation. Unexpectedly, GCs have anti-apoptotic effects in keratinocytes by inducing the expression of anti-apoptotic and repressing pro-apoptotic genes. Consequently, GCs treatment blocked UV-induced apoptosis of keratinocytes. GCs have a profound effect on wound healing by inhibiting cell motility and the expression of pro-angiogenic factor VEGF. They play an important role in tissue remodeling and scar formation by suppressing the expression of TGF-beta-1 and -2, MMP1, 2, 9 and 10 and inducing TIMP-2. Finally, GCs promote terminal stages of epidermal differentiation while simultaneously inhibiting the early stages. These results provide new insights into the beneficial and adverse effects of GCs in epidermis, defining the participating genes and mechanisms that coordinate the cellular responses important for GC-based therapies. Human epidermal keratinocytes are grown in delipidized, phenolphtalein-free medium and left as controls or treated with 0.1μM dexamethasone. Time course of treated and parallel control samples over a 72 hr period was performed twice with independent batches of cells.
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Project description:Human epidermal keratinocytes were treated with 25 ng.ml EphB2 or EFNA4, both as-Fc conjugates (Sigma). Human epidermal keratinocytes are treated with 25 ng/ml EphB2 or EFNA4 Fc conjugates in a 48hr time course.
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Project description:BMP signalling is a potent regulator of skin morphogenesis, homeostasis and remodelling. However, molecular mechanisms underlying its involvement in regulating gene expression programs in keratinocytes and fibroblasts remain largely unknown. We analyzed the effect of BMP4 tratment on gene expression programs in human primary epidermal keratinocyte and dermal fibroblasts cultures. We identified specific changes in gene expression programs for each cell type. The primary human epidermal keratinocytes and dermal fibroblasts were treated with recombinant BMP4 or solvent control for 8 hours to reveal early and intermediate response genes. The RNA was isolated and used for micro-array analysis. Changes in gene expression programs were analyzed for each cell type and were compared between cell types.
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