Home LiteratureArticle Details
PMID: 9464988 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S.

Extracellular signal-regulated kinase and p38 subgroups of mitogen-activated protein kinases regulate inducible nitric oxide synthase and tumor necrosis factor-alpha gene expression in endotoxin-stimulated primary glial cultures.

The Journal of neuroscience : the official journal of the Society for Neuroscience ·Vol. 18 ·No. 5 ·1998-03-01 ·Pages 1633-41

Bhat NR, Zhang P, Lee JC, Hogan EL

Abstract

Tumor necrosis factor-alpha (TNFalpha) and nitric oxide (NO), the product of inducible NO synthase (iNOS), mediate inflammatory and immune responses in the CNS under a variety of neuropathological situations. They are produced mainly by "activated" astrocytes and microglia, the two immune regulatory cells of the CNS. In this study we have examined the regulation of TNFalpha and iNOS gene expression in endotoxin-stimulated primary glial cultures, focusing on the role of mitogen-activated protein (MAP) kinase cascades. The bacterial lipopolysaccharide (LPS) was able to activate extracellular signal-regulated kinase (ERK) and p38 kinase subgroups of MAP kinases in microglia and astrocytes. ERK activation was sensitive to PD98059, the kinase inhibitor that is specific for ERK kinase. The activity of p38 kinase was inhibited by SB203580, a member of the novel class of cytokine suppressive anti-inflammatory drugs (CSAIDs), as revealed by blocked activation of the downstream kinase, MAP kinase-activated protein kinase-2. The treatment of glial cells with either LPS alone (microglia) or a combination of LPS and interferon-gamma (astrocytes) resulted in an induced production of NO and TNFalpha. The two kinase inhibitors, at micromolar concentrations, individually suppressed and, in combination, almost completely blocked glial production of NO and the expression of iNOS and TNFalpha, as determined by Western blot analysis. Reverse transcriptase-PCR analysis showed changes in iNOS mRNA levels that paralleled iNOS protein and NO while indicating a lack of effect of either of the kinase inhibitors on TNFalpha mRNA expression. The results demonstrate key roles for ERK and p38 MAP kinase cascades in the transcriptional and post-transcriptional regulation of iNOS and TNFalpha gene expression in endotoxin-activated glial cells.

MeSH Terms
Animals Calcium-Calmodulin-Dependent Protein Kinases/antagonists & inhibitors,metabolism Cells, Cultured Enzyme Activation Enzyme Induction Enzyme Inhibitors/pharmacology Female Gene Expression Regulation, Enzymologic Lipopolysaccharides/pharmacology Microglia/enzymology,metabolism Mitogen-Activated Protein Kinase 1 Mitogen-Activated Protein Kinases Nitric Oxide/antagonists & inhibitors Nitric Oxide Synthase/biosynthesis,genetics Nitric Oxide Synthase Type II Polymerase Chain Reaction Rats Rats, Sprague-Dawley Tumor Necrosis Factor-alpha/biosynthesis,genetics p38 Mitogen-Activated Protein Kinases
Chemicals
Enzyme Inhibitors Lipopolysaccharides Tumor Necrosis Factor-alpha Nitric Oxide Nitric Oxide Synthase Nitric Oxide Synthase Type II Nos2 protein, rat Calcium-Calmodulin-Dependent Protein Kinases Mitogen-Activated Protein Kinase 1 Mitogen-Activated Protein Kinases p38 Mitogen-Activated Protein Kinases
Authors & Affiliations
4 authors, click to expand affiliations / ORCID
Bhat N R
Department of Neurology, Medical University of South Carolina, Charleston, South Carolina 29425, USA.
Zhang P
Lee J C
Hogan E L
References (64)
64 references, click to expand
  1. Inducible nitric oxide synthase in Theiler's murine encephalomyelitis virus infection.
    J Virol. 1997 Apr;71(4):3228-35 PMID: 9060686
  2. Macrophages and inflammation in the central nervous system.
    Trends Neurosci. 1993 Jul;16(7):268-73 PMID: 7689770
  3. Mitogen-activated protein kinases activate the serine/threonine kinases Mnk1 and Mnk2.
    EMBO J. 1997 Apr 15;16(8):1909-20 PMID: 9155017
  4. Promoter of the mouse gene encoding calcium-independent nitric oxide synthase confers inducibility by interferon gamma and bacterial lipopolysaccharide.
    J Exp Med. 1993 Jun 1;177(6):1779-84 PMID: 7684434
  5. Hemopoietic growth factors with the exception of interleukin-4 activate the p38 mitogen-activated protein kinase pathway.
    J Biol Chem. 1997 Feb 7;272(6):3296-301 PMID: 9013568
  6. Cytokines and metabolic dysfunction after severe head injury.
    J Neurotrauma. 1994 Oct;11(5):447-72 PMID: 7861440
  7. A synthetic inhibitor of the mitogen-activated protein kinase cascade.
    Proc Natl Acad Sci U S A. 1995 Aug 15;92(17):7686-9 PMID: 7644477
  8. Protein tyrosine kinase inhibitors suppress the production of nitric oxide in mixed glia, microglia-enriched or astrocyte-enriched cultures.
    Brain Res. 1996 Aug 5;729(1):102-9 PMID: 8874881
  9. The role of nitric oxide in multiple sclerosis.
    J Mol Med (Berl). 1997 Mar;75(3):174-86 PMID: 9106074
  10. The expression of myristoylated alanine-rich C-kinase substrate in oligodendrocytes is developmentally regulated.
    Dev Neurosci. 1995;17(4):256-63 PMID: 8575345
  11. The regulation of AP-1 activity by mitogen-activated protein kinases.
    J Biol Chem. 1995 Jul 14;270(28):16483-6 PMID: 7622446
  12. An increase in intracellular cyclic AMP modulates nitric oxide production in IFN-gamma-treated macrophages.
    J Immunol. 1997 Jan 15;158(2):897-904 PMID: 8993009
  13. Identification of a common nucleotide sequence in the 3'-untranslated region of mRNA molecules specifying inflammatory mediators.
    Proc Natl Acad Sci U S A. 1986 Mar;83(6):1670-4 PMID: 2419912
  14. Microglia: a sensor for pathological events in the CNS.
    Trends Neurosci. 1996 Aug;19(8):312-8 PMID: 8843599
  15. Nitric oxide in neuronal degeneration.
    Proc Soc Exp Biol Med. 1996 Jan;211(1):33-40 PMID: 8594616
  16. Activation of mitogen-activated protein kinases in oligodendrocytes.
    J Neurochem. 1996 May;66(5):1986-94 PMID: 8780027
  17. Interactions of the nervous and immune systems in development, normal brain homeostasis, and disease.
    FASEB J. 1995 May;9(8):611-8 PMID: 7768352
  18. Regulation of cytokine-inducible nitric oxide synthase in cardiac myocytes and microvascular endothelial cells. Role of extracellular signal-regulated kinases 1 and 2 (ERK1/ERK2) and STAT1 alpha.
    J Biol Chem. 1996 Jan 12;271(2):1111-7 PMID: 8557638
  19. Pharmacological profile of SB 203580, a selective inhibitor of cytokine suppressive binding protein/p38 kinase, in animal models of arthritis, bone resorption, endotoxin shock and immune function.
    J Pharmacol Exp Ther. 1996 Dec;279(3):1453-61 PMID: 8968371
  20. How MAP kinases are regulated.
    J Biol Chem. 1995 Jun 23;270(25):14843-6 PMID: 7797459
  21. Nitric oxide-mediated mitochondrial damage in the brain: mechanisms and implications for neurodegenerative diseases.
    J Neurochem. 1997 Jun;68(6):2227-40 PMID: 9166714
  22. Synthesis of nitric oxide in CNS glial cells.
    Trends Neurosci. 1993 Aug;16(8):323-8 PMID: 7691008
  23. Cytoplasmic regulation of mRNA function: the importance of the 3' untranslated region.
    Cell. 1993 Jul 16;74(1):9-14 PMID: 7687524
  24. Role of CSB/p38/RK stress response kinase in LPS and cytokine signaling mechanisms.
    J Leukoc Biol. 1996 Feb;59(2):152-7 PMID: 8603987
  25. Up-regulation of protein kinase C-epsilon promotes the expression of cytokine-inducible nitric oxide synthase in RAW 264.7 cells.
    J Biol Chem. 1996 Dec 13;271(50):32028-33 PMID: 8943252
  26. Production of tumor necrosis factor and other cytokines by astrocytes stimulated with lipopolysaccharide or a neurotropic virus.
    Proc Natl Acad Sci U S A. 1989 Aug;86(16):6348-52 PMID: 2474832
  27. Cytokine- and endotoxin-induced nitric oxide synthase in rat astroglial cultures: differential modulation by angiotensin II.
    J Neurochem. 1997 Mar;68(3):935-44 PMID: 9048738
  28. MAPKs: new JNK expands the group.
    Trends Biochem Sci. 1994 Nov;19(11):470-3 PMID: 7855889
  29. Nitric oxide synthase type II mRNA stability is translation- and transcription-dependent.
    J Neurochem. 1996 Oct;67(4):1766-9 PMID: 8858964
  30. Transcriptional regulation by extracellular signals: mechanisms and specificity.
    Cell. 1995 Jan 27;80(2):199-211 PMID: 7834740
  31. Induction of nitric oxide synthase by cyclic AMP in rat vascular smooth muscle cells.
    J Clin Invest. 1994 Feb;93(2):543-9 PMID: 7509342
  32. Cytokine production by human fetal microglia and astrocytes. Differential induction by lipopolysaccharide and IL-1 beta.
    J Immunol. 1993 Apr 1;150(7):2659-67 PMID: 8454848
  33. Cytokine mRNA expression in postischemic/reperfused myocardium.
    Am J Pathol. 1995 Feb;146(2):419-28 PMID: 7856752
  34. Nitric oxide. A macrophage product responsible for cytostasis and respiratory inhibition in tumor target cells.
    J Exp Med. 1989 May 1;169(5):1543-55 PMID: 2497225
  35. Macrophage nitric oxide synthase gene: two upstream regions mediate induction by interferon gamma and lipopolysaccharide.
    Proc Natl Acad Sci U S A. 1993 Oct 15;90(20):9730-4 PMID: 7692452
  36. AU-rich elements: characterization and importance in mRNA degradation.
    Trends Biochem Sci. 1995 Nov;20(11):465-70 PMID: 8578590
  37. Regulation of cachectin biosynthesis occurs at multiple levels.
    Prog Clin Biol Res. 1990;349:229-40 PMID: 2204931
  38. Interactive effects of the tumor necrosis factor promoter and 3'-untranslated regions.
    J Immunol. 1991 Mar 15;146(6):1843-8 PMID: 1848573
  39. Requirement for transcription factor IRF-1 in NO synthase induction in macrophages.
    Science. 1994 Mar 18;263(5153):1612-5 PMID: 7510419
  40. Parallel signal processing among mammalian MAPKs.
    Trends Biochem Sci. 1995 Mar;20(3):117-22 PMID: 7709430
  41. Activation of Stat1 and subsequent transcription of inducible nitric oxide synthase gene in C6 glioma cells is independent of interferon-gamma-induced MAPK activation that is mediated by p21ras.
    FEBS Lett. 1997 May 12;408(1):33-8 PMID: 9180263
  42. Thrombin activates mitogen-activated protein kinase in primary astrocyte cultures.
    J Cell Physiol. 1995 Nov;165(2):417-24 PMID: 7593220
  43. PHAS-I as a link between mitogen-activated protein kinase and translation initiation.
    Science. 1994 Oct 28;266(5185):653-6 PMID: 7939721
  44. Activation of JNK/SAPK in primary glial cultures: II. Differential activation of kinase isoforms corresponds to their differential expression.
    Neurochem Res. 1998 Feb;23(2):219-25 PMID: 9475517
  45. MNK1, a new MAP kinase-activated protein kinase, isolated by a novel expression screening method for identifying protein kinase substrates.
    EMBO J. 1997 Apr 15;16(8):1921-33 PMID: 9155018
  46. Immunologic NO synthase: elevation in severe AIDS dementia and induction by HIV-1 gp41.
    Science. 1996 Dec 13;274(5294):1917-21 PMID: 8943206
  47. Tumor necrosis factor-alpha potentiates glutamate neurotoxicity in human fetal brain cell cultures.
    Dev Neurosci. 1994;16(3-4):172-9 PMID: 7705222
  48. Inducible nitric oxide synthase following hypoxia in rat cultured glial cells.
    Brain Res. 1996 Nov 4;738(2):319-22 PMID: 8955528
  49. A protein kinase involved in the regulation of inflammatory cytokine biosynthesis.
    Nature. 1994 Dec 22-29;372(6508):739-46 PMID: 7997261
  50. Insulin-dependent stimulation of protein synthesis by phosphorylation of a regulator of 5'-cap function.
    Nature. 1994 Oct 27;371(6500):762-7 PMID: 7935836
  51. In vivo expression of inducible nitric oxide synthase in experimentally induced neurologic diseases.
    Proc Natl Acad Sci U S A. 1993 Apr 1;90(7):3024-7 PMID: 7681993
  52. Sounding the alarm: protein kinase cascades activated by stress and inflammation.
    J Biol Chem. 1996 Oct 4;271(40):24313-6 PMID: 8798679
  53. Differential regulation of cytokine-induced major histocompatibility complex class II expression and nitric oxide release in rat microglia and astrocytes by effectors of tyrosine kinase, protein kinase C, and cAMP.
    J Neuroimmunol. 1997 Apr;74(1-2):19-29 PMID: 9119973
  54. Expression of inducible nitric oxide synthase causes delayed neurotoxicity in primary mixed neuronal-glial cortical cultures.
    Neuropharmacology. 1994 Nov;33(11):1425-30 PMID: 7532825
  55. Astrocytes in infectious and immune-mediated diseases of the central nervous system.
    FASEB J. 1993 Oct;7(13):1226-32 PMID: 8405808
  56. Nitric oxide synthase in models of focal ischemia.
    Stroke. 1997 Jun;28(6):1283-8 PMID: 9183363
  57. Multiple sclerosis: TNF revisited, with promise.
    Nat Med. 1995 Mar;1(3):211-4 PMID: 7585035
  58. Cloning and expression of inducible nitric oxide synthase from rat astrocytes.
    J Neurosci Res. 1994 Feb 15;37(3):406-14 PMID: 7513765
  59. Roles of tyrosine kinases in the regulation of nitric oxide synthesis in murine liver cells: modulation of NF-kappa B activity by tyrosine kinases.
    Hepatology. 1997 Apr;25(4):913-9 PMID: 9096597
  60. Neuronal (type I) nitric oxide synthase regulates nuclear factor kappaB activity and immunologic (type II) nitric oxide synthase expression.
    Proc Natl Acad Sci U S A. 1997 Mar 18;94(6):2676-80 PMID: 9122255
  61. Transcriptional regulation of the tumor necrosis factor alpha gene.
    Immunobiology. 1995 Jul;193(2-4):210-6 PMID: 8530145
  62. Bacterial lipopolysaccharide plus interferon-gamma elicit a very fast inhibition of a Ca2+-dependent nitric-oxide synthase activity in human astrocytoma cells.
    J Biol Chem. 1997 Mar 21;272(12):7582-5 PMID: 9065411
  63. Activation of the inducible form of nitric oxide synthase in the brains of patients with multiple sclerosis.
    Proc Natl Acad Sci U S A. 1995 Dec 19;92(26):12041-5 PMID: 8618840
  64. p38 mitogen-activated protein kinase down-regulates nitric oxide and up-regulates prostaglandin E2 biosynthesis stimulated by interleukin-1beta.
    J Biol Chem. 1997 Mar 21;272(12):8083-9 PMID: 9065483
Article Info
Journal
The Journal of neuroscience : the official journal of the Society for Neuroscience
Abbr.
J Neurosci
ISSN
0270-6474
Published
1998-03-01
Pages
1633-41
Language
English
Region
United States
NLM ID
8102140
PMCID
PMC6792636
Subset
IM
Grants
NINDS NIH HHS · NS31767 · United States
External Links
PubMed source PMC Full Text View journal details
Analysis Services
Analysis Services

Contact

No. 2 Wenbo Road, Zhangqiu District, Jinan, Shandong

Qilu Normal University · Genelibs Bioinformatics Lab

750 Shunhua Rd, Jinan

2F, Bldg F, University Science Park

Tel: 0531-88819269

WeChat Official Account

Follow our WeChat subscription account for real-time updates and the latest in medical and biological research.


Business Email

E-mail: [email protected]

Copyright © 2026.Genelibs Jinan Honghui Biotechnology Co., Ltd. 鲁ICP备20007159号-2