Adenylyl cyclase activation and overexpression inhibited TGF-beta- and angiotensin II-stimulated alpha-SMA expression and collagen synthesis in adult rat cardiac fibroblasts.
Does adenylyl cyclase activation and overexpression inhibit the transformation of adult rat cardiac fibroblasts to myofibroblasts and reduce collagen synthesis?
Increasing adenylyl cyclase expression and cAMP generation in cardiac fibroblasts inhibits their transformation to myofibroblasts and reduces collagen synthesis, highlighting a potential strategy to attenuate cardiac fibrosis.
Transformation of fibroblasts to myofibroblasts, characterized by expression of alpha-smooth muscle actin (alpha-SMA) and production of extracellular matrix (ECM) components, is a key event in connective tissue remodeling. Approaches to inhibit this transformation are needed in tissues, such as the heart, where excessive ECM production by cardiac fibroblasts (CFs) causes fibrosis, myocardial stiffening, and cardiac dysfunction. We tested whether adenylyl cyclase (AC) activation (increased cAMP levels) modulates the transformation of adult rat CF to myofibroblasts, as assessed by immunofluorescent microscopy, immunoblotting, and collagen synthesis. A 24-h incubation of CF with TGF-beta or angiotensin II increased alpha-SMA expression, which was inhibited by the AC agonist forskolin and a cAMP analog that activates protein kinase A. Treatment with forskolin blunted serum-, TGF-beta-, and angiotensin II-stimulated collagen synthesis. CFs engineered to overexpress type 6 AC had enhanced forskolin-promoted cAMP formation, greater inhibition by forskolin of TGF-beta-stimulated alpha-SMA expression, and a decrease in the EC(50) of forskolin to reduce serum-stimulated collagen synthesis. The AC stimulatory agonist adrenomedullin inhibited collagen synthesis in CF that overexpressed AC6 but not in controls. Thus, AC stimulation blunts collagen synthesis and, in parallel, the transformation of adult rat CF to myofibroblasts. AC overexpression enhances these effects, "uncovering" an inhibition by adrenomedullin. These findings implicate cAMP as an inhibitor of ECM formation by means of blockade of the transformation of CF to myofibroblasts and suggest that increasing AC expression, thereby enhancing cAMP generation through stimulation of receptors expressed on CF, could provide a means to attenuate and prevent cardiac fibrosis and its sequelae.
Swaney et al. (Wed,) conducted a other in Cardiac fibrosis (in vitro model). Adenylyl cyclase activation and overexpression vs. Control (untreated or non-overexpressing fibroblasts) was evaluated on Transformation of fibroblasts to myofibroblasts (alpha-SMA expression) and collagen synthesis. Adenylyl cyclase activation and overexpression inhibited TGF-beta- and angiotensin II-stimulated alpha-SMA expression and collagen synthesis in adult rat cardiac fibroblasts.