Gadolinium chloride inhibits lipopolysaccharide-induced mortality and in vivo prostaglandin E2 release By splenic macrophages.
The monocytic phagocytic system, consisting primarily of tissue macrophages of the liver and spleen, produces prostaglandin E(2) (PGE(2)), a modulator of the septic response. Macrophages are known to internalize gadolinium chloride (GD), a lanthanide metal, which inhibits phagocytic function. Thus we studied the effect of in vivo GD on lipopolysacchride (LPS)-induced mortality and on LPS-stimulated PGE(2) release by cultured splenic macrophages. GD (7 mg/kg intravenously) given on the two days prior to LPS challenge (30 mg/kg intravenously) completely prevented the uniform mortality in rats. This protective effect was transient since rechallenge with LPS 10 days later was uniformly lethal. Previous work in this laboratory has established a critical role of arginine concentration on macrophage behavior in vitro. Therefore, to establish culture conditions reflective of the milieu within the portal venous system, alanine and arginine levels were measured in the portal and hepatic veins of normal and endotoxemic (LPS, 10 mg/kg intraperitoneally) rats. In contrast to alanine levels, which were not altered by endotoxemia, there was a reduction of arginine concentrations from a range of 50 to 250 micromol/L in normal rats to a range of 10 to 50 micromol/L after LPS challenge. Consequently subsequent in vitro assays of splenic macrophage secretory behavior were performed in concentrations of 1200 micromol/L arginine (in standard RPMI-1640), as well as in concentrations reflective of physiologic arginine levels (10 and 100 micromol/L in modified RPMI-1640). Rat splenic macrophages harvested after two consecutive days of either in vivo saline or GD injection (7 mg/kg intravenously) were stimulated with LPS (0.025 to 2.5 microg/ml). At 72 hours of culture, the release of PGE(2) by splenic macrophages from GD-treated rats was significantly (P <0.0001) reduced at all LPS concentrations. Increased PGE(2) production was not present when the splenic macrophages were cultured in the supraphysiologic arginine (1200 micromol/L) concentration. The results demonstrate the relevance of physiologic arginine concentrations in cell culture studies and suggest that the protection conferred by GD against septic mortality may be related to downregulation of the release of immunosuppressive PGE(2) by the monocytic phagocytic system.
Roland
C R
CR
Department of Surgery, Washington University School of Medicine, St. Louis, MO 63110, USA.
Nakafusa
Y
Y
Flye
M W
MW
D000409:Alanine / Q000097:blood; D000818:Animals; D000893:Anti-Inflammatory Agents / Q000008:administration & dosage / Q000627:therapeutic use*; D001120:Arginine / Q000097:blood / Q000494:pharmacology; D002470:Cell Survival / Q000187:drug effects; D002478:Cells, Cultured; D019610:Cytoprotection / Q000187:drug effects; D015232:Dinoprostone / Q000037:antagonists & inhibitors / Q000378:metabolism*; D015536:Down-Regulation; D019446:Endotoxemia / Q000097:blood; D005682:Gadolinium / Q000008:administration & dosage / Q000627:therapeutic use*; D006503:Hepatic Veins; D007275:Injections, Intravenous; D008070:Lipopolysaccharides / Q000009:adverse effects* / Q000037:antagonists & inhibitors; D008264:Macrophages / Q000187:drug effects* / Q000378:metabolism; D008297:Male; D010587:Phagocytosis / Q000187:drug effects; D011169:Portal Vein; D051381:Rats; D017207:Rats, Sprague-Dawley; D018805:Sepsis / Q000378:metabolism; D013154:Spleen / Q000166:cytology / Q000187:drug effects*
D000893:Anti-Inflammatory Agents; D008070:Lipopolysaccharides; D001120:Arginine; D005682:Gadolinium; D015232:Dinoprostone; D000409:Alanine; C038958:gadolinium chloride