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Canadian Journal of Anesthesia, Vol 45, 246-252, Copyright © 1998 by Canadian Anesthesiologists' Society

ARTICLES

Dantrolene inhibits nitric oxide synthase in rat alveolar macrophages treated with lipopolysaccharide and interferon-gamma

CY Li, TC Chou, CC Wu, CS Wong, ST Ho, MH Yen and YA Ding
Department of Anesthesiology, Graduate Institute of Medical Sciences, Tri-Service General Hospital, National Defense Medical Center, Taipei, Taiwan, Republic of China. cyli@ndmc1.ndmctsgh.edu.tw

PURPOSE: To examine the effects of dantrolene on nitric oxide (NO) production and on the activity and protein expression of inducible nitric oxide synthase (iNOS) induced by lipopolysaccharide (LPS) plus interferon-gamma (IFN gamma) in rat alveolar macrophages. METHODS: Pulmonary alveolar macrophages isolated from Sprague-Dawley rats were used. After incubation of macrophages with dantrolene (1 to 100 microM) and LPS (1 microgram.ml-1) and IFN-gamma (100 u.ml-1) for 24 hr, the cell-free medium was removed for measuring the nitrite and tumour necrosis factor-alpha (TNF-alpha) levels by Griess reaction and ELISA kit, respectively. The harvested macrophages were also used to determine the activity of iNOS by using the conversion of [3H]-L-arginine to [3H]-L-citrulline method. Protein expression of iNOS was detected by Western blot analysis. RESULTS: In rats alveolar macrophages, (i) dantrolene (1 to 100 microM) caused a dose-dependent suppression of the production of nitrite and TNF-alpha induced by LPS (1 microgram.ml-1) plus IFN-gamma (100 u.ml-1) and (ii) dantrolene (100 microM) inhibited the activity (by 37 +/- 5%, P < 0.01) and protein expression (by 39 +/- 12%, P < 0.01) of iNOS in response to LPS plus IFN-gamma. CONCLUSION: Dantrolene inhibits NO production as well as the activity and expression of iNOS in alveolar macrophages treated with LPS plus IFN-gamma, which may be associated with the reduction of TNF-alpha production.




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Copyright © 1998 by the Canadian Anesthesiologists' Society.