Abstract:
BACKGROUND:Nuclear factor kappa B (NF-kappaB) has been shown to play an important role in regulating the expression of many genes involved in cell survival, immunity and in the inflammatory processes. NF-kappaB activation upregulates inducible nitric oxide synthase leading to enhanced nitric oxide production during an inflammatory response. NF-kappaB activation is regulated by distinct kinase pathways independent of inhibitor of kappaB kinase (IKK). Here, we examine the role of protein kinase C isoforms and janus activated kinase 2 (JAK2) activation in NF-kappaB activation and LPS-stimulated NO production. METHODS:Murine RAW 264.7 macrophages were treated with lipopolysaccharide (LPS), Phorbol 12-myristate 13-acetate (PMA) and a combination of LPS and PMA in the presence or absence of various inhibitors of PKC isoforms and JAK2. Nuclear translocation of the NF-kappaB p65 subunit, was assessed by Western blot analysis whilst NO levels were assessed by Greiss assay. RESULTS:LPS-stimulated NO production was attenuated by PMA whilst PMA alone did not affect NO release. These effects were associated with changes in p65 nuclear translocation. The PKCalpha, beta, gamma, delta and zeta inhibitor Gö 6983 (Go) had no effect on LPS-induced NO release. In contrast, Bisindolymalemide I (Bis), a PKC alpha, betaI, betaII, gamma, delta and epsilon isoform inhibitors completely inhibited LPS-stimulated NO production without affecting p65 nuclear translocation. Furthermore, a partial inhibitory effect on LPS-induced NO release was seen with the JAK2 inhibitor AG-490 and the p38 MAPK inhibitor SB 203850. CONCLUSION:The results further define the role of NF-kappaB in LPS stimulated NO production in RAW macrophages. The data support a function for PKCepsilon, JAK2 and p38 MAPK in NF-kappaB activation following p65 nuclear import.
journal_name
J Inflamm (Lond)journal_title
Journal of inflammation (London, England)authors
Jones E,Adcock IM,Ahmed BY,Punchard NAdoi
10.1186/1476-9255-4-23subject
Has Abstractpub_date
2007-11-24 00:00:00pages
23issn
1476-9255pii
1476-9255-4-23journal_volume
4pub_type
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journal_title:Journal of inflammation (London, England)
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journal_title:Journal of inflammation (London, England)
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