cDNA cloning of canine adrenomedullin and its gene expression in the heart and blood vessels in endotoxin shock

Abstract

Adrenomedullin (AM) is a potent vasorelaxing peptide isolated from human pheochromocytoma. It is proposed that AM dilates the blood vessels by autocrine or paracrine mechanisms. Our investigation concerned whether the vasodilating function of AM accounts for the hypotension occurring in endotoxin shock. The effect of endotoxin shock on AM gene expression in the cardiovascular system was probed using a canine model. The cDNA encoding the AM precursor in dogs was isolated from cDNA libraries of the adrenal gland and was sequenced. The canine AM precursor is constituted of 188 amino acids and its AM consists of 52 amino acids, similar to human and porcine AM. The dogs were injected intravenously with 2 mg/kg of lipopolysaccharide (LPS) after being anesthetized and intubated. After 4 h, we collected whole blood, the large and small blood vessels, and the heart from our canine model of endotoxin shock, and extracted RNA from those tissues for Northern blot analysis. In the endotoxin shock model, the mRNA levels of the AM had increased in almost all of the blood vessels. These results suggest that AM may dilate the blood vessels systemically, even if AM acts as a local modulator of vascular tone. In addition, the plasma concentrations of the AM were high enough to allow for AM to act as a vasodilating hormone. Consequently, AM may be one of the factors facilitating severe hypotension complicated with endotoxin shock.