Effects of adrenomedullin on aldosterone-induced cell proliferation in rat cardiac fibroblasts

Aldosterone induces cardiac remodeling in cardiovascular diseases by stimulating the proliferation, production and secretion of collagen in fibroblasts. It also stimulates vascular smooth muscle cells to produce and secrete adrenomedullin (ADM), which has a cytoprotective effect against cardiovascular damage. We examined the effect of aldosterone on ADM production and secretion in rat cardiac fibroblasts, and the effect of ADM on aldosterone-stimulated fibroblast proliferation to observe the interaction between endogenous ADM and aldosterone. We detected ADM produced and secreted from cultured cardiac fibroblasts and the intracellular cAMP level by radioimmunoassay; evaluated cell proliferation by the level of [H-3]-thymine incorporation; measured preproADM gene expression by reverse transcriptase polymerase chain reaction (RT-PCR); and monitored extracellular signal related kinase (ERK) activity by the phosphorylation of myelin basic protein in the presence of [gamma-P-32] ATP. Our results showed that aldosterone-stimulated secretion of ADM and its mRNA expression were concentration-dependent, which could be inhibited by the specific antagonist of mineralocorticoid receptor, spironolactone. In contrast, ADM inhibited aldosterone-induced fibroblast proliferation and ERK activity. Treatment with ADM(24-50) (a new antagonist of specific ADM receptors) and calcitonin gene-related peptide (CGRP)(8-37) (the antagonist of CGRP receptor type 1), to attenuate the action of endogenous ADM, reinforced the aldosterone-induced proliferation and inhibited the intracellular cAMP production stimulated by aldosterone. Thiorphan, an inhibitor of ADM degradation, inhibited the [H-3]-thymine incorporation and reinforced the intracellular cAMP level induced by aldosterone. We reach the conclusion that aldosterone stimulates rat cardiac fibroblasts to produce and secrete ADM, which in turn regulates the proliferation-induced effects of aldosterone in these cells. (C) 2004 Elsevier B.V. All rights reserved.