title: Endotoxin tolerance is associated with altered GTP-binding protein function.
abstract: Previous studies have suggested that guanine nucleotide regulatory (G) proteins modulate endotoxin-stimulated peritoneal macrophage arachidonic acid (AA) metabolism. Endotoxin-stimulated metabolism of AA by peritoneal macrophages is decreased in endotoxin tolerance (Rogers et al. Prostaglandins 31: 639-650, 1986). These observations led to a study of G protein function and AA metabolism by peritoneal macrophages in endotoxin tolerance. Endotoxin tolerance was induced by the administration of sublethal doses of endotoxin. AA metabolism was assessed by measurement of thromboxane B2 (TxB2), a cyclooxygenase metabolite. NaF (5 mM), an activator of G proteins, significantly stimulated TxB2 synthesis in control macrophages from 7.7 +/- 0.2 to 19.1 +/- 0.6 (SE) ng/ml (P less than 0.05) at 2 h and was partially inhibited by pertussis toxin, suggesting a G protein-dependent mechanism. Salmonella enteritidis endotoxin (50 micrograms/ml) stimulated a similar increase in TxB2 levels (23 +/- 0.4 ng/ml, P less than 0.05). In contrast to control macrophages, macrophages from endotoxin-tolerant rats stimulated with either NaF or S. enteritidis endotoxin had TxB2 levels that were only 30 and 2% of the respective stimulated control cells. Basal guanosine-triphosphatase (GTPase) activity (33 +/- 6 pmol.mg-1.min-1) in endotoxin-tolerant macrophage membranes was significantly lower (P less than 0.05) than control basal activity (158 +/- 5 pmol.mg-1.min-1). This suppression of macrophage GTPase activity was apparent 48 h after the first in vivo sublethal endotoxin injection (100 micrograms/kg ip). The reduced GTPase activity paralleled in vitro cellular hyporesponsiveness to endotoxin-stimulated TxB2 production.(ABSTRACT TRUNCATED AT 250 WORDS)
authors.affiliation: Department of Physiology, Medical University of South Carolina, Charleston 29425.
authors.forename: K A
authors.initials: KA
authors.lastname: Coffee
authors.forename: P V
authors.initials: PV
authors.lastname: Halushka
authors.forename: S H
authors.initials: SH
authors.lastname: Ashton
authors.forename: G E
authors.initials: GE
authors.lastname: Tempel
authors.forename: W C
authors.initials: WC
authors.lastname: Wise
authors.forename: J A
authors.initials: JA
authors.lastname: Cook
mesh_terms: D000818:Animals; D016718:Arachidonic Acid / Q000378:metabolism; D002470:Cell Survival / Q000187:drug effects; D004361:Drug Tolerance; D004731:Endotoxins / Q000633:toxicity*; D005260:Female; D020558:GTP Phosphohydrolases / Q000378:metabolism; D019204:GTP-Binding Proteins / Q000378:metabolism*; D066298:In Vitro Techniques; D008050:Lipid A / Q000494:pharmacology; D008264:Macrophages / Q000187:drug effects* / Q000378:metabolism; D051381:Rats; D012969:Sodium Fluoride / Q000494:pharmacology; D013929:Thromboxane B2 / Q000096:biosynthesis
chemical_list: D004731:Endotoxins; D008050:Lipid A; D016718:Arachidonic Acid; D013929:Thromboxane B2; D012969:Sodium Fluoride; D020558:GTP Phosphohydrolases; D019204:GTP-Binding Proteins