Andrographolide suppresses the expression of inducible nitric oxide synthase in macrophage and restores the vasoconstriction in rat aorta treated with lipopolysaccharide

1 We investigated whether andrographolide, a diterpenoid lactone found at Andrographis paniculata, influences the induction of the inducible nitric oxide synthase (iNOS) in RAW264.7 cells activated by bacterial endotoxin (LPS), as well as in the rats with endotoxic shock and in aortic rings treated with LPS. 2 Incubation of RAW264.7 cells with andrographolide (1 to 50 mu M) inhibited the LPS (1 mu g ml(-1))-induced nitrite accumulation in concentration- and time-dependent manners. Maximum inhibition was observed when andrographolide was added together with LPS and decreased progressively as the interval between andrographolide and LPS was increased to 20 h. 3 Western blot analysis demonstrated that iNOS expression was markedly attenuated in the presence of andrographolide for 6-24 h, suggesting that andrographolide inhibited iNOS protein induction. 4 Thoracic aorta incubation with LPS (300 ng ml(-1)) for 5 h in vitro exhibited a significant decrease in the maximal contractile response to phenylephrine (10(-9)-10(-5) M). Andrographolide (30 mu M) restored the contractile response to control level. 5 In anaesthetized rats, LPS (10 mg kg(-1), i.v.) caused a fall in mean arterial blood pressure (MAP) from 116 +/- 4 to 77 +/- 5 mmHg. The presser effect of phenylephrine (10 mu g ml(-1), i.v.) was also significantly reduced at 30, 60, 120 and 180 min after LPS injection. In contrast, animals pretreated with andrographolide (1 mg kg(-1), i.v., 20 min prior to LPS) maintained a significantly higher MAP when compared to LPS-rats given with vehicle. Administration of andrographolide 60 min after LPS caused a increase in MAP and significantly reversed the reduction of the presser response to phenylephrine. 6 Our results indicated that andrographolide inhibits nitrite synthesis by suppressing expression of iNOS protein in vitro. And, this inhibition of iNOS synthesis may contribute to the beneficial haemodynamic effects of andrographolide in endotoxic shock.