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KMID : 0378120030300010021
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Chungnam Medical Journal
2003 Volume.30 No. 1 p.21 ~ p.35
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Transcriptional Regulation of Inducible Nitric Oxide Synthase Gene in C6 Glial Cell
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Bae Jin-Young
Hur Gang-Min
Bea So-Hyun
Park Ji-Sun
Lee Choong-Jae
Lee Jea-Heun
Seok Jeong-Ho
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Abstract
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To investigate transcriptional regulation of iNOS gene by LPS and cytokines, the production of NO, expression of iNOS mRNA and protein, binding activity of nuclear factor-kappa B(NF-kB), and promoter activity of iNOS gene were examined in rat C6 glial cells. LPS, interferon-gamma(IFN-γ), and tumor necrosis factor-alpha (TNF-α) stimulated the production of NO, which was increased synergistically by co-treatment. By the treatment of LPS, iNOS mRNA expression was initiated at 1 h, markedly increased by 3 h, and decreased gradually afterward. iNOS mRNA expression was markedly enhanced by mixture of LPS, IFN-γ and TNF-α. iNOS protein synthesis was increased by the treatment of mixture LPS and cytokine mixture. Treatment of LPS stimulated NF-kB activation, and the activation reached to the maximum level at 30 min, and the treatment of mixture of LPS and cytokines increased the activation. To determine the effect of NF-kB binding activity on iNOS promoter activation, CAT assay was performed. iNOS promoter activity was increased by the treatment with LPS for 5.5 h, and further increased by the combined treatment with LPS and cytokines.
These results suggest that NF-kB activation by LPS and cytokines may play a significant role in the induction of the iNOS gene.
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KEYWORD
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iNOS, C6 glial cell, NF-kB
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