RABBIT BRAIN CONTAINS AN ENDOGENOUS INHIBITOR OF ENDOTHELIUM-DEPENDENT RELAXATION

1. Supernatants prepared from the rabbit brain, lung and liver caused an endothelium-dependent and volume-related contraction of the phenylephrine-pretreated rabbit aorta and inhibited relaxation due to acetylcholine (ACh). 2. Perfusion in situ of the rabbit lung or liver with Krebs solution substantially reduced or removed the endothelium-dependent inhibitor. Spectrophotometric analysis revealed the presence of substantial amounts of haemoglobin (1.8-2.1 μM) in these organ supernatants. 3. Supernatants prepared from the Krebs-perfused rabbit brain retained the ability to contract the phenylephrine-pretreated rabbit aorta and to inhibit relaxation due to ACh and substance P (SP). Rabbit brain supernatant did not reduce the vasodilator effect of sodium nitroprusside (NP) or nitric oxide (NO). 4. Rabbit brain supernatant contained low (<0.35 μM) concentrations of haemoglobin. 5. The inhibitory effect of rabbit brain supernatant was reversed by L-arginine (500 μM) but not D-arginine (500 μM). 6. The inhibitor of endothelium-dependent vasodilatation present in rabbit brain was not removed by dialysis (24 h, 4°C) but was partially precipitated by ammonium sulphate (30% w/v). 7. Rabbit brain contains an endogenous inhibitor of vascular NO biosynthesis. The identity of this inhibitor is not known although it seems likely to be a large peptide or protein.