Cannabinoid signalling in TNF-alpha induced IL-8 release.
The molecular events mediating the immunomodulatory properties of cannabinoids have remained largely unresolved. We have therefore investigated the molecular mechanism(s) through which R-(+)-[2,3-dihydro-5-methyl-3-[(morpholinyl)methyl] pyrrolo[1,2,3-de]-1,4-benzoxazinyl]-(1-napthanlenyl) methanone (WIN55212-2) modulate production of interleukin-8 (IL-8) in HT-29 cells. Release of IL-8 induced by tumor necrosis factor-alpha (TNF-alpha) was determined by enzyme-linked immunosorbent assay (ELISA). Changes in expression of inhibitory kappa B (IkappaB) were monitored by Western blotting and activation of nuclear factor-kappa B (NF-kappaB) was determined in electrophoretic mobility shift assay (EMSAs). TNF-alpha induced release of IL-8 was inhibited by WIN55212-2 which also blocked the degradation of IkappaB-alpha and activation of NF-kappaB induced by TNF-alpha. These data provide strong evidence that WIN55212-2 may modulate IL-8 release by negatively regulating the signaling cascade leading to the activation of NF-kappaB. These findings highlight a potential mechanism for the immunomodulatory properties of cannabinoids and contribute towards acquiring a clear understanding of the role of cannabinoids in inflammation.
Mormina
Maria E
ME
School of Life Sciences, University of Hertfordshire, Faculty of Health and Human Sciences, College Lane, Hatfield, Herts AL10 9AB, United Kingdom.
Thakur
Shori
S
Molleman
Areles
A
Whelan
Cliff J
CJ
Baydoun
Anwar R
AR
D048588:Benzoxazines; D015153:Blotting, Western; D063386:Cannabinoid Receptor Agonists; D002186:Cannabinoids / Q000378:metabolism; D003513:Cycloheximide / Q000494:pharmacology; D015853:Cysteine Proteinase Inhibitors / Q000494:pharmacology; D003609:Dactinomycin / Q000494:pharmacology; D004305:Dose-Response Relationship, Drug; D004797:Enzyme-Linked Immunosorbent Assay; D019073:HT29 Cells; D006801:Humans; D020672:I-kappa B Proteins / Q000378:metabolism; D016209:Interleukin-8 / Q000378:metabolism*; D007976:Leupeptins / Q000494:pharmacology; D009025:Morpholines / Q000494:pharmacology; D016328:NF-kappa B / Q000378:metabolism; D009281:Naphthalenes / Q000494:pharmacology; D011500:Protein Synthesis Inhibitors / Q000494:pharmacology; D043882:Receptors, Cannabinoid / Q000502:physiology*; D015398:Signal Transduction / Q000187:drug effects / Q000502:physiology*; D014409:Tumor Necrosis Factor-alpha / Q000494:pharmacology*; D015854:Up-Regulation / Q000187:drug effects
D048588:Benzoxazines; D063386:Cannabinoid Receptor Agonists; D002186:Cannabinoids; D015853:Cysteine Proteinase Inhibitors; D020672:I-kappa B Proteins; D016209:Interleukin-8; D007976:Leupeptins; D009025:Morpholines; D016328:NF-kappa B; D009281:Naphthalenes; D011500:Protein Synthesis Inhibitors; D043882:Receptors, Cannabinoid; D014409:Tumor Necrosis Factor-alpha; D003609:Dactinomycin; C070417:(3R)-((2,3-dihydro-5-methyl-3-((4-morpholinyl)methyl)pyrrolo-(1,2,3-de)-1,4-benzoxazin-6-yl)(1-naphthalenyl))methanone; D003513:Cycloheximide; C072553:benzyloxycarbonylleucyl-leucyl-leucine aldehyde