Morphine inhibits NF-kappaB nuclear binding in human neutrophils and monocytes by a nitric oxide-dependent mechanism

• Published in: Anesthesiology (Philadelphia) 2000-06, Vol.92 (6), p.1677-1684
• Main Authors: Welters, I D, Menzebach, A, Goumon, Y, Cadet, P, Menges, T, Hughes, T K, Hempelmann, G, Stefano, G B
• Format: Article
• Language: English
• Subjects: Acetylcysteine - pharmacology; Analgesics, Opioid - pharmacology; Cell Nucleus - drug effects; Cell Nucleus - metabolism; Enzyme Inhibitors - pharmacology; Flow Cytometry; Humans; In Vitro Techniques; Lipopolysaccharides; Male; Monocytes - drug effects; Monocytes - metabolism; Morphine - pharmacology; Naloxone - pharmacology; Narcotic Antagonists - pharmacology; Neutrophils - drug effects; Neutrophils - metabolism; NF-kappa B - drug effects; NF-kappa B - metabolism; NG-Nitroarginine Methyl Ester - pharmacology; Nitric Oxide - physiology; Nitric Oxide Synthase - antagonists & inhibitors; Protein Binding; Receptors, Opioid, mu - drug effects; Receptors, Opioid, mu - metabolism
• ISSN: 0003-3022

The transcription factor NF-kappaB plays a pivotal role in gene expression of inflammatory mediators such as cytokines or adhesion molecules. NF-kappaB-mediated transcriptional activation of these genes is inhibited by nitric oxide (NO) in a variety of cells, including monocytes. Morphine mediates NO release in a naloxone antagonizable manner in monocytes and neutrophils. The influence of morphine on NF-kappaB activation was investigated in a whole-blood flow cytometric assay. A specific antibody against the p65 subunit of NF-kappaB was used and detected by fluoresceine-isothiocyanate-labeled anti-immunoglobulin G. Nuclei were stained with propidium iodide. Leukocyte subpopulations were evaluated by gating on neutrophils and monocytes. The median fluorescence channel was determined. Different morphine concentrations (50 nm, 50 microm, 1 mm) and incubation intervals (10-150 min) were used. Morphine inhibits lipopolysaccharide-induced NF-kappaB nuclear binding in human blood neutrophils and monocytes in a time-, concentration-, and naloxone-sensitive-dependent manner. Similar effects were achieved with the NO donor S-nitroso-N-acetyl-pencillamine and the antioxidant N-acetyl-cysteine. The NO synthase inhibitors Nomega-nitro-l-arginine-methyl-esther and Nomega-nitro-l-arginine completely abolished the morphine-induced attenuation of NF-kappaB nuclear binding, demonstrating that the inhibitory action is mediated by NO release. Morphine causes immunosuppression, at least in part, via the NO-stimulated depression of NF-kappaB nuclear binding.