Evaluation of iNOS‐dependent and independent mechanisms of the microvascular permeability change induced by lipopolysaccharide

Subcutaneous injection of lipopolysaccharide (LPS) increases plasma leakage in mouse skin. Pretreatment with LPS conditions mice tolerant to the LPS‐induced plasma leakage. Nitric oxide (NO) has been suggested to be involved in these LPS effects. A specific role of inducible NO synthase (iNOS) was investigated in the LPS‐induced plasma leakage using iNOS deficient mice. Plasma leakage in mouse skin was measured by the local accumulation of Pontamine sky blue at the site of subcutaneous injection of LPS (Sal. typhimurium). LPS (100–400 μg site −1 ) produced a dose‐related increase in dye leakage in both iNOS deficient and wild‐type mice with about 40% less dye leakage in iNOS deficient mice. Indomethacin (5 mg kg −1 ), N‐[‐2‐cyclohexyloxy]‐4‐nitrophenyl methanesulphonamide (NS‐398) (1 mg kg −1 ), diphenhydramine (10 mg kg −1 ) and anti‐TNF‐α antibody (dilution 1 : 400, 10 ml kg −1 ) inhibited the LPS‐induced dye leakage in both iNOS deficient and wild‐type mice, whereas N G ‐nitro‐ L ‐arginine methyl ester ( L ‐NAME) (10 mg kg −1 ) or aminoguanidine (10 mg kg −1 ) inhibited that in wild‐type but not in iNOS deficient mice. Pretreatment with LPS (0.15 mg kg −1 i.p.) 4 h before decreased the LPS‐induced dye leakage in wild‐type but not in iNOS deficient mice. LPS pretreatment increased serum corticosterone levels in both mice, while it increased the serum nitrate/nitrite levels in wild‐type but not in iNOS deficient mice. These studies indicate that an increase in vascular permeability induced by LPS is mediated by NO produced by iNOS, eicosanoids, histamine and TNF‐α. The tolerance against LPS‐induced vascular permeability change may be mediated by iNOS induction but not by an increased release of endogenous corticosteroids.British Journal of Pharmacology (2000) 130 , 90–94; doi: 10.1038/sj.bjp.0703277