Abstract 078: Functional Significance of the Chymase/Ang-(1-12) Pathway on Cardiomyocytes

Abstract

Angiotensin-(1-12) [Ang-(1-12)] functions in rodents and humans as a tissue substrate for the direct generation of Ang II via chymase. Since its direct cardiac effect have not been studied, the importance of this renin-independent mechanism for Ang II paracrine/intracrine actions in modulating cardiac contractility were determined in freshly isolated myocytes from 11 normal SD rats. Systolic amplitude (SA), peak velocity of shortening (dL/dtmax), the peak velocity of relengthening (dR/dtmax), and changes in the peak calcium transient ([Ca2+]i) were evaluated before and following exposure to Ang II (10-6 M), Ang-(1-12) delivered alone (range: 2x10-6 to 4x10-6 M) or after 1 h incubation with human recombinant chymase (10 μg protein/mL at 37°C). Both Ang II and the mixture of Ang-(1-12) with chymase elicited positive inotropic responses in freshly isolated cardiac myocytes associated with significant increases in peak systolic [Ca2+]i (Figure) while superfusion of Ang-(1-12) alone elicited an increase in dL/dtmax without significant changes in [Ca2+]i. The increases in contractility elicited by Ang II or the Ang-(1-12)/chymase mixture were abolished by prior exposure of the myocytes to losartan (10-5 M) or the chymase inhibitor chymostatin (8x10-5 M). We conclude that in single adult rat myocytes Ang-(1-12) stimulates contractile function through a chymase mediated action and by mechanisms that implicate a paracrine/intracrine activation of intracellular calcium.