Involvement of mitogen-activated protein kinase homologues in the regulation of lipopolysaccharide-mediated induction of cyclo-oxygenase-2 but not nitric oxide synthase in RAW 264.7 macrophages

Andrew Paul; Ana Cuenda; Clare E. Bryant; Jo Murray; Edwin R. Chilvers; Philip Cohen; Gwyn W. Gould; Robin Plevin

doi:10.1016/S0898-6568(99)00018-2

Involvement of mitogen-activated protein kinase homologues in the regulation of lipopolysaccharide-mediated induction of cyclo-oxygenase-2 but not nitric oxide synthase in RAW 264.7 macrophages

Andrew Paul^*, Ana Cuenda, Clare E. Bryant, Jo Murray, Edwin R. Chilvers, Philip Cohen, Gwyn W. Gould, Robin Plevin

^*Corresponding author for this work

Strathclyde Institute Of Pharmacy And Biomedical Sciences

Research output: Contribution to journal › Article › peer-review

102 Citations (Scopus)

Abstract

In RAW 264.7 macrophages lipopolysaccharide (LPS) stimulated the activation of p42 and p44 MAP kinases and their upstream activator mitogen-activated protein (MAP) kinase kinase (MAPKK), and induced the 69-kDa isoform of cyclo-oxygenase-2 (COX-2) and the 130-kDa isoform of nitric oxide synthase (iNOS). PD 098059, a specific inhibitor of the activation of MAPKK, prevented LPS-mediated activation of MAPKK IC₅₀ = 3.0 ± 0.1 μM, n = 3 and p42/44 MAP kinases and substantially reduced the induction of COX-2 by approximately 40%-70%, but was without effect upon the induction of iNOS. In parallel, LPS also stimulated the activation of p38 MAP kinase and the MAPKAP kinase-2, a downstream target of p38 MAP kinase. SB 203580, a specific inhibitor of p38 MAP kinase prevented the activation of p38 MAP kinase IC₅₀ = 3.3 ± 1.4 μM, n = 3 and MAPKAP kinase-2 by LPS and reduced the induction of COX-2 by approximately 50-90%, with no significant effect upon iNOS expression. These studies indicate the involvement of both the classical p42/44 MAP kinases and p38 MAP kinase in the regulation of COX-2 but not iNOS induction following exposure to LPS. Copyright (C) 1999 Elsevier Science Inc.

Original language	English
Pages (from-to)	491-497
Number of pages	7
Journal	Cellular Signalling
Volume	11
Issue number	7
DOIs	https://doi.org/10.1016/S0898-6568(99)00018-2
Publication status	Published - 1 Jul 1999

Funding

The authors thank Alan R. Saltiel (Parke Davis, USA) for the kind gift of PD 098059, John Lee and Peter Young (Smith Kline & Beecham, USA) for the gift of SB 203580, Dr. L. Cowell (Amersham International, Bucks., UK) for the BIOTRAK™ MAP kinase assay kit, Gary Johnson (Denver, CO, USA) for the plasmids expressing histidine-tagged MAP kinase and Prof. C. Marshall (ICR, London, UK) for the plasmid expressing GST-MAPKAP kinase-2. This work was funded by grants from the Medical Research Council and the Wellcome Trust (to R. P. and G. W. G.) and by the Royal Society (to P. C.). C. E. B. is a Wellcome Trust Research Fellow in Vetinary Science. E. R. C. is a Wellcome Trust Senior Research Fellow in Clinical Science. G. W. G. is a 1992 Lister Fellow.

Keywords

COX-2
iNOS
lipopolysaccharide
MAP kinase
PD 098059
SB 203580
mitogen activated protein kinase
nitric oxide synthase
enzyme activation

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Paul, A., Cuenda, A., Bryant, C. E., Murray, J., Chilvers, E. R., Cohen, P., Gould, G. W., & Plevin, R. (1999). Involvement of mitogen-activated protein kinase homologues in the regulation of lipopolysaccharide-mediated induction of cyclo-oxygenase-2 but not nitric oxide synthase in RAW 264.7 macrophages. Cellular Signalling, 11(7), 491-497. https://doi.org/10.1016/S0898-6568(99)00018-2

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title = "Involvement of mitogen-activated protein kinase homologues in the regulation of lipopolysaccharide-mediated induction of cyclo-oxygenase-2 but not nitric oxide synthase in RAW 264.7 macrophages",

abstract = "In RAW 264.7 macrophages lipopolysaccharide (LPS) stimulated the activation of p42 and p44 MAP kinases and their upstream activator mitogen-activated protein (MAP) kinase kinase (MAPKK), and induced the 69-kDa isoform of cyclo-oxygenase-2 (COX-2) and the 130-kDa isoform of nitric oxide synthase (iNOS). PD 098059, a specific inhibitor of the activation of MAPKK, prevented LPS-mediated activation of MAPKK IC50 = 3.0 ± 0.1 μM, n = 3 and p42/44 MAP kinases and substantially reduced the induction of COX-2 by approximately 40%-70%, but was without effect upon the induction of iNOS. In parallel, LPS also stimulated the activation of p38 MAP kinase and the MAPKAP kinase-2, a downstream target of p38 MAP kinase. SB 203580, a specific inhibitor of p38 MAP kinase prevented the activation of p38 MAP kinase IC50 = 3.3 ± 1.4 μM, n = 3 and MAPKAP kinase-2 by LPS and reduced the induction of COX-2 by approximately 50-90%, with no significant effect upon iNOS expression. These studies indicate the involvement of both the classical p42/44 MAP kinases and p38 MAP kinase in the regulation of COX-2 but not iNOS induction following exposure to LPS. Copyright (C) 1999 Elsevier Science Inc.",

keywords = "COX-2, iNOS, lipopolysaccharide, MAP kinase, PD 098059, SB 203580, mitogen activated protein kinase, nitric oxide synthase, enzyme activation",

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Paul, A, Cuenda, A, Bryant, CE, Murray, J, Chilvers, ER, Cohen, P, Gould, GW & Plevin, R 1999, 'Involvement of mitogen-activated protein kinase homologues in the regulation of lipopolysaccharide-mediated induction of cyclo-oxygenase-2 but not nitric oxide synthase in RAW 264.7 macrophages', Cellular Signalling, vol. 11, no. 7, pp. 491-497. https://doi.org/10.1016/S0898-6568(99)00018-2

Involvement of mitogen-activated protein kinase homologues in the regulation of lipopolysaccharide-mediated induction of cyclo-oxygenase-2 but not nitric oxide synthase in RAW 264.7 macrophages. / Paul, Andrew; Cuenda, Ana; Bryant, Clare E. et al.
In: Cellular Signalling, Vol. 11, No. 7, 01.07.1999, p. 491-497.

Research output: Contribution to journal › Article › peer-review

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AU - Paul, Andrew

AU - Cuenda, Ana

AU - Bryant, Clare E.

AU - Murray, Jo

AU - Chilvers, Edwin R.

AU - Cohen, Philip

AU - Gould, Gwyn W.

AU - Plevin, Robin

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AB - In RAW 264.7 macrophages lipopolysaccharide (LPS) stimulated the activation of p42 and p44 MAP kinases and their upstream activator mitogen-activated protein (MAP) kinase kinase (MAPKK), and induced the 69-kDa isoform of cyclo-oxygenase-2 (COX-2) and the 130-kDa isoform of nitric oxide synthase (iNOS). PD 098059, a specific inhibitor of the activation of MAPKK, prevented LPS-mediated activation of MAPKK IC50 = 3.0 ± 0.1 μM, n = 3 and p42/44 MAP kinases and substantially reduced the induction of COX-2 by approximately 40%-70%, but was without effect upon the induction of iNOS. In parallel, LPS also stimulated the activation of p38 MAP kinase and the MAPKAP kinase-2, a downstream target of p38 MAP kinase. SB 203580, a specific inhibitor of p38 MAP kinase prevented the activation of p38 MAP kinase IC50 = 3.3 ± 1.4 μM, n = 3 and MAPKAP kinase-2 by LPS and reduced the induction of COX-2 by approximately 50-90%, with no significant effect upon iNOS expression. These studies indicate the involvement of both the classical p42/44 MAP kinases and p38 MAP kinase in the regulation of COX-2 but not iNOS induction following exposure to LPS. Copyright (C) 1999 Elsevier Science Inc.

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KW - iNOS

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KW - MAP kinase

KW - PD 098059

KW - SB 203580

KW - mitogen activated protein kinase

KW - nitric oxide synthase

KW - enzyme activation

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SN - 0898-6568

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ER -

Involvement of mitogen-activated protein kinase homologues in the regulation of lipopolysaccharide-mediated induction of cyclo-oxygenase-2 but not nitric oxide synthase in RAW 264.7 macrophages

Abstract

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Keywords

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