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A Mechanism for the Up-regulation of the IL-8 Gene Expression in Keratinocytes by All-trans Retinoic Acid / 대한피부과학회지
Article em Ko | WPRIM | ID: wpr-113620
Biblioteca responsável: WPRO
ABSTRACT
BACKGROUND: Retinoic acid (RA) has been reported to induce the up-regulation of inflammatory cytokines such as IL-1, TNF-alpha and IL-8 in dermal fibroblasts and keratinocytes. There is no evidence to support a direct interaction between the RA-mediated transcriptional machinery and IL-8 gene transcription. OBJECTIVE: The aim of this study is to clarify the mechanism of the up-regulation of IL-8 in keratinocytes by RA. METHODS: The IL-1, IL-8, TNF-alpha and MCP-1 mRNA expressions in HaCaT cells stimulated by RA were measured by quantitative RT-PCR. The effects of a NF-kappaB inhibitor and IL-1 receptor antagonist (ra) on the IL-8 mRNA expression were measured by quantitative RT-PCR. Electrophoretic motility shift assay (EMSA) was conducted on the RA-stimulated HaCaT cells that were or were not treated with NF-kappaB inhibitor to measure the NF-kappaB binding activity in each group. The phospho-IkappaB activity in the HaCaT cells after stimulation with RA was also measured by Western blotting. RESULTS: An up-regulation of the IL-8 gene expression by RA was demonstrated in the HaCaT cells. The inhibition assay revealed the involvement of the NF-kappaB binding site of the IL-8 gene in the RA-enhanced promoter activity. EMSA demonstrated that RA enhanced the formation of the DNA-NF-kappaB complex. There was no evidence to support IL-1 as an intermediate stimulus between the RA-mediated transcriptional machinery and IL-8 gene transcription. Western blot analysis revealed increased phospho-IkappaB activity in the HaCaT cells after stimulation with RA. CONCLUSION: Our result suggested that the IL-8 gene expression of HaCaT cells after RA stimulation is caused by the activation of IKK and the dissociation of IkappaB from NF-kappaB and the transcription of NF-kappaB in the nucleus.
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Texto completo: 1 Índice: WPRIM Assunto principal: Tretinoína / Sítios de Ligação / RNA Mensageiro / Expressão Gênica / Queratinócitos / Regulação para Cima / Western Blotting / Citocinas / Interleucina-8 / NF-kappa B Idioma: Ko Revista: Korean Journal of Dermatology Ano de publicação: 2009 Tipo de documento: Article
Texto completo: 1 Índice: WPRIM Assunto principal: Tretinoína / Sítios de Ligação / RNA Mensageiro / Expressão Gênica / Queratinócitos / Regulação para Cima / Western Blotting / Citocinas / Interleucina-8 / NF-kappa B Idioma: Ko Revista: Korean Journal of Dermatology Ano de publicação: 2009 Tipo de documento: Article