title: Neurotoxicity of soluble macrophage products in vitro--influence of dexamethasone.
abstract: When macrophage conditioned medium is added to neurons in vitro, there is a loss of cell membrane integrity, a loss of cell processes, and a large increase in apoptotic neurons. We tested the influence of a potent anti-inflammatory steroid on the interaction between macrophages and neurons. Dexamethasone was applied to macrophages in culture for 24 h while the culture was stimulated with lipopolysaccharide and hypoxia. Conditioned medium was collected after dexamethasone was removed. The dexamethasone pretreated medium was not toxic to hippocampal neurons in contrast to medium from stimulated macrophages not treated with steroid. The dexamethasone effect was concentration dependent. Pretreatment of macrophages with indomethacin and transforming growth factor beta had similar but less impressive effects when compared to dexamethasone. The effect of dexamethasone may have been mediated by inhibiting the synthesis or release of neurotoxic macrophage protein(s), as a combination of medium from steroid pretreated macrophages with medium from nontreated macrophages was not neuroprotective. The toxin(s) did not appear to be tumor necrosis factor alpha or arginase. A role for most neutral proteases was also excluded. We also assessed the consequence of stressing neurons with a mild hypoxic exposure immediately prior to conditioned medium application. Medium from dexamethasone-treated macrophages did not exaggerate hypoxic neuronal injury, unlike medium from non-dexamethasone-treated macrophages. It did not, however, block the exaggerating effect when coapplied in equal volume with medium from nontreated macrophages. Dexamethasone at 100 nM had no impact when applied directly to neurons while they were being exposed to conditioned medium. This in vitro protection by dexamethasone may be relevant to the demonstrated benefit of glucocorticoids in selected brain and spinal cord conditions. Suspicion of a potential link between this in vitro finding and in vivo CNS injury justifies an assessment of more specific agents acting on macrophage protein synthesis or secretion.
authors.affiliation: Department of Pediatrics, Queen's University, Kingston, Ontario, Canada.
authors.forename: M P
authors.initials: MP
authors.lastname: Flavin
authors.forename: L T
authors.initials: LT
authors.lastname: Ho
authors.forename: K
authors.initials: K
authors.lastname: Coughlin
mesh_terms: D000818:Animals; D000894:Anti-Inflammatory Agents, Non-Steroidal / Q000494:pharmacology; D001119:Arginase / Q000494:pharmacology; D015687:Cell Hypoxia / Q000187:drug effects; D002478:Cells, Cultured; D017077:Culture Media, Conditioned / Q000737:chemistry / Q000494:pharmacology; D003907:Dexamethasone / Q000494:pharmacology*; D004305:Dose-Response Relationship, Drug; D004586:Electrophoresis; D005938:Glucocorticoids / Q000494:pharmacology*; D006624:Hippocampus / Q000166:cytology; D007213:Indomethacin / Q000494:pharmacology; D017737:Macrophages, Peritoneal / Q000166:cytology / Q000187:drug effects / Q000378:metabolism*; D051379:Mice; D008815:Mice, Inbred Strains; D009474:Neurons / Q000166:cytology / Q000187:drug effects*; D009498:Neurotoxins / Q000378:metabolism*; D012995:Solubility; D016212:Transforming Growth Factor beta / Q000494:pharmacology; D014409:Tumor Necrosis Factor-alpha / Q000494:pharmacology
chemical_list: D000894:Anti-Inflammatory Agents, Non-Steroidal; D017077:Culture Media, Conditioned; D005938:Glucocorticoids; D009498:Neurotoxins; D016212:Transforming Growth Factor beta; D014409:Tumor Necrosis Factor-alpha; D003907:Dexamethasone; D001119:Arginase; D007213:Indomethacin