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R(+)-Methanandamide Induces Cyclooxygenase-2 Expression in Human Neuroglioma Cells via a Non-cannabinoid Receptor-Mediated Mechanism

https://doi.org/10.1006/bbrc.2001.5518Get rights and content

Abstract

Cannabinoids affect prostaglandin (PG) formation in the central nervous system through as yet unidentified mechanisms. Using H4 human neuroglioma cells, the present study investigates the effect of R(+)-methanandamide (metabolically stable analogue of the endocannabinoid anandamide) on the expression of the cyclooxygenase-2 (COX-2) enzyme. Incubation of cells with R(+)-methanandamide was accompanied by concentration-dependent increases in COX-2 mRNA, COX-2 protein, and COX-2-dependent PGE2 synthesis. Moreover, treatment of cells with R(+)-methanandamide in the presence of interleukin-1β led to an overadditive induction of COX-2 expression. The stimulatory effect of R(+)-methanandamide on COX-2 expression was mimicked by the structurally unrelated cannabinoid Δ9-tetrahydrocannabinol. Stimulation of both COX-2 mRNA expression and subsequent PGE2 synthesis by R(+)-methanandamide was not affected by the selective CB1 receptor antagonist AM-251 or the Gi/o protein inactivator pertussis toxin. Enhancement of COX-2 expression by R(+)-methanandamide was paralleled by time-dependent phosphorylations of p38 mitogen-activated protein kinase (MAPK) and p42/44 MAPK. Consistent with the activation of both kinases, R(+)-methanandamide-induced COX-2 mRNA expression and PGE2 formation were abrogated in the presence of specific inhibitors of p38 MAPK (SB203580) and p42/44 MAPK activation (PD98059). Together, our results demonstrate that R(+)-methanandamide induces COX-2 expression in human neuroglioma cells via a cannabinoid receptor-independent mechanism involving activation of the MAPK pathway. In conclusion, induction of COX-2 expression may represent a novel mechanism by which cannabinoids mediate PG-dependent effects within the central nervous system.

References (56)

  • D.G. Deutsch et al.

    Enzymatic synthesis and degradation of anandamide, a cannabinoid receptor agonist

    Biochem. Pharmacol.

    (1993)
  • N. Futaki et al.

    NS-398, a new anti-inflammatory agent, selectively inhibits prostaglandin G/H synthase/cyclooxygenase (COX-2) activity in vitro

    Prostaglandins

    (1994)
  • S.J. Gatley et al.

    123I-labeled AM251: A radioiodinated ligand which binds in vivo to mouse brain cannabinoid CB1 receptors

    Eur. J. Pharmacol.

    (1996)
  • D. Hwang et al.

    Inhibition of the expression of inducible cyclooxygenase and proinflammatory cytokines by sesquiterpene lactones in macrophages correlates with the inhibition of MAP kinases

    Biochem. Biophys. Res. Commun.

    (1996)
  • J.L. Dean et al.

    p38 mitogen-activated protein kinase regulates cyclooxygenase-2 mRNA stability and transcription in lipopolysaccharide-treated human monocytes

    J. Biol. Chem.

    (1999)
  • Z. Guan et al.

    Interleukin-1β-induced cyclooxygenase-2 expression requires activation of both c-Jun NH2-terminal kinase and p38 MAPK signal pathways in rat renal mesangial cells

    J. Biol. Chem.

    (1998)
  • H. Niiro et al.

    MAP kinase pathways as a route for regulatory mechanisms of IL-10 and IL-4 which inhibit COX-2 expression in human monocytes

    Biochem. Biophys. Res. Commun.

    (1998)
  • A. Cuenda et al.

    SB 203580 is a specific inhibitor of a MAP kinase homologue which is stimulated by cellular stresses and interleukin-1

    FEBS Lett.

    (1995)
  • M. Perez-Reyes et al.

    Antagonism of marihuana effects by indomethacin in humans

    Life Sci.

    (1991)
  • T. Yamaguchi et al.

    Behavioral suppression induced by cannabinoids is due to activation of the arachidonic acid cascade in rats

    Brain Res.

    (2001)
  • D.W. Pate et al.

    Effects of topical anandamides on intraocular pressure in normotensive rabbits

    Life Sci.

    (1996)
  • M. Wartmann et al.

    The MAP kinase signal transduction pathway is activated by the endogenous cannabinoid anandamide

    FEBS Lett.

    (1995)
  • M. Tsujii et al.

    Alterations in cellular adhesion and apoptosis in epithelial cells overexpressing prostaglandin endoperoxide synthase-2

    Cell

    (1995)
  • C. Sanchez et al.

    Δ9-Tetrahydrocannabinol induces apoptosis in C6 glioma cells

    FEBS Lett.

    (1998)
  • M. Yu et al.

    Synthesis of prostaglandin E2 ethanolamide from anandamide by cyclooxygenase-2

    J. Biol. Chem.

    (1997)
  • K.R. Kozak et al.

    Oxygenation of the endocannabinoid, 2-arachidonylglycerol, to glyceryl prostaglandins by cyclooxygenase-2

    J. Biol. Chem.

    (2000)
  • R. Mechoulam et al.

    A total synthesis of d1 delta-1 tetrahydrocannabinol, the active constituent of hashish

    J. Am. Chem. Soc.

    (1965)
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      Further support for this assumption may be derived from our finding that exogenous PGE2 did not reduce COX-2 expression, but rather increased COX-2 protein levels in H4 cells. Interestingly, anandamide as well as its hydrolysis-stable analog, R(+)-methanandamide, both having structural similarities with alkamides, have previously been shown to cause upregulation of COX-2 expression through a mechanism involving increased ceramide synthesis and activation of mitogen-activated protein kinases [10–13]. However, in both cases induction of COX-2 expression was associated with increased COX-2 activity resulting in enhanced levels of PGE2 in cell culture supernatants.

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    Abbreviations used: COX, cyclooxygenase; cAMP, adenosine 3′,5′-cyclic monophosphate; ERK, extracellular signal-regulated kinase; HRP, horseradish peroxidase; IL-1β, interleukin-1β; MAPK, mitogen-activated protein kinase; PG, prostaglandin; Δ9-THC, Δ9-tetrahydrocannabinol.

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