Role of nitric oxide and prostaglandins in the potentiating effects of calcitonin gene-related peptide on lipopolysaccharide-induced interleukin-6 release from mouse peritoneal macrophages.
Previous data from our laboratory have shown that calcitonin gene-related peptide (CGRP) has a potentiating effect on lipopolysaccharide-(LPS) induced interleukin-6 (IL-6) release from mouse macrophages. However, the mechanism of this effect was not clear. Since the nitric oxide (NO) and prostaglandins (PGs) induced by LPS might modulate IL-6 release, we examined whether NO and PGs were also involved in the potentiating effect of rat CGRP (rCGRP) on LPS-induced IL-6 release from mouse macrophages. The IL-6 level in the medium was measured by enzyme-linked immunosorbent assay. Accumulation of NO was assessed by measuring the presence of nitrite by the Greiss reaction. PGI2 was assessed by measuring the formation of 6-keto-prostaglandin F1alpha (6-keto-PGF1alpha) by radioimmunoassay. The results showed that the potentiating effect of rCGRP (0.1 nm) on LPS-induced IL-6 release was significantly inhibited by either 100 micrometers NG-monomethyl-L-arginine acetate (L-NMMA; an inhibitor of NO synthase) or 10 micrometers indomethacin (an inhibitor of cyclo-oxygenase). The LPS-induced NO and PGI2 production from these cells was increased significantly by rCGRP at 0.01-10 nm in a concentration-dependent manner, which was blocked by L-NMMA and indomethacin. These results suggest that rCGRP enhances the NO production elicited by LPS and subsequently increases the PGs production which is involved in the potentiating effect of rCGRP on LPS-induced IL-6 release from the peritoneal macrophages in the mouse.
Tang
Y
Y
Institute of Vascular Medicine, Third Hospital, Beijing Medical University, Beijing, China.
Han
C
C
Wang
X
X
D000704:Analysis of Variance; D000818:Animals; D015740:Calcitonin Gene-Related Peptide / Q000494:pharmacology*; D002478:Cells, Cultured; D016861:Cyclooxygenase Inhibitors / Q000494:pharmacology; D004305:Dose-Response Relationship, Drug; D011464:Epoprostenol / Q000378:metabolism*; D007213:Indomethacin / Q000494:pharmacology; D015850:Interleukin-6 / Q000378:metabolism*; D008070:Lipopolysaccharides / Q000494:pharmacology; D017737:Macrophages, Peritoneal / Q000187:drug effects* / Q000276:immunology / Q000378:metabolism; D008297:Male; D051379:Mice; D008807:Mice, Inbred BALB C; D009569:Nitric Oxide / Q000378:metabolism*; D019001:Nitric Oxide Synthase / Q000037:antagonists & inhibitors; D051381:Rats; D011994:Recombinant Proteins / Q000494:pharmacology; D019323:omega-N-Methylarginine / Q000494:pharmacology
D016861:Cyclooxygenase Inhibitors; D015850:Interleukin-6; D008070:Lipopolysaccharides; D011994:Recombinant Proteins; D019323:omega-N-Methylarginine; D009569:Nitric Oxide; D011464:Epoprostenol; D019001:Nitric Oxide Synthase; D015740:Calcitonin Gene-Related Peptide; D007213:Indomethacin