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Tumor necrosis factor α (TNF-α)-induced prostaglanding E2 release is mediated by the activation of cyclooxygenase-2 (COX-2)transcription via NFκB in human gingival fibroblasts

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Abstract

Nuclear factor kappaB (NFκB) is a transcription factor and plays a key role in the expression of several genes involved in the inflammatory process. Cyclooxygenase (COX) is the key regulatory enzyme of the prostaglandin/eicosanoid synthetic pathway. COX-2 is a highly inducible enzyme by proinflammatory cytokines, of which gene expression is regulated by NFκB. TNF-α is a pro-inflammatory cytokine. In this paper, we investigated the involvement of NFκB on TNF-α-mediated prostaglandin E2 (PGE2) release and COX-2 gene expression in human gingival fibroblasts (HGF). TNF-α- induced PGE2 release and COX-2 mRNA accumulation in a time- and concentration-dependent manner in HGF. The results of transient transfection assays using a chimeric construct of the human COX-2 promoter (nts –1432 ∼ +59) ligated to a luciferase reporter gene indicated that TNF-α stimulated the transcriptional activity ∼ 1.4-fold. Gel mobility shift assays with a radiolabelled COX-2-NFκB oligonucleotide (nts –223 to –214) revealed an increase in the binding of nuclear proteins from TNF-α-stimulated HGF. The COX-2-NFκB DNA-protein complex disappeared after treatment with pyrrolidine dithiocarbamate (PDTC; an antioxidant) or herbimycin A (a tyrosine kinase inhibitor). PDTC and herbimycin A attenuated TNF-α-stimulated PGE2 release. These results suggest that NFκB transcription factor is a key regulator of COX-2 expression in TNF-α-induced PGE2 production, which is mediated through a tyrosine kinase pathway in HGF.

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Authors and Affiliations

  1. Department of Pharmacology, Research Institute of Oral Science, Nihon University School of Dentistry at Matsudo, Matsudo, Chiba, 271-8587, Japan

    Sumi Nakao

  2. Department of Periodontology, Research Institute of Oral Science, Nihon University School of Dentistry at Matsudo, Matsudo, Chiba, 271-8587, Japan

    Yorimasa Ogtata

  3. Department of Endodontics, Research Institute of Oral Science, Nihon University School of Dentistry at Matsudo, Matsudo, Chiba, 271-8587, Japan

    Emi Shimizu & Muneyoshi Yamazaki

  4. Department of Physiology, Research Institute of Oral Science, Nihon University School of Dentistry at Matsudo, Matsudo, Chiba, 271-8587, Japan

    Shunsuke Furuyama & Hiroshi Sugiya

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  1. Sumi Nakao
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  4. Muneyoshi Yamazaki
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  5. Shunsuke Furuyama
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Nakao, S., Ogtata, Y., Shimizu, E. et al. Tumor necrosis factor α (TNF-α)-induced prostaglanding E2 release is mediated by the activation of cyclooxygenase-2 (COX-2)transcription via NFκB in human gingival fibroblasts. Mol Cell Biochem 238, 11–18 (2002). https://doi.org/10.1023/A:1019927616000

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