RESUMEN

AIMS: Pathological tissue remodelling by myofibroblast contraction is a hallmark of cardiac fibrosis. Myofibroblasts differentiate from cardiac fibroblasts under the action of transforming growth factor-ß1 (TGF-ß1), which is secreted into the extracellular matrix as a large latent complex. Integrin-mediated traction forces activate TGF-ß1 by inducing a conformational change in the latent complex. The mesenchymal integrins αvß5 and αvß3 are expressed in the heart, but their role in the activation of TGF-ß1 remains elusive. Here, we test whether targeting αvß5 and αvß3 integrins reduces latent TGF-ß1 activation by cardiac fibroblasts with the goal to prevent the formation of α-smooth muscle actin (α-SMA)-expressing cardiac myofibroblasts and their contribution to fibrosis. METHODS AND RESULTS: Using a porcine model of induced right ventricular fibrosis and pro-fibrotic culture conditions, we show that integrins αvß5 and αvß3 are up-regulated in myofibroblast-enriched fibrotic lesions and differentiated cultured human cardiac myofibroblasts. Both integrins autonomously contribute to latent TGF-ß1 activation and myofibroblast differentiation, as demonstrated by function-blocking peptides and antibodies. Acute blocking of both integrins leads to significantly reduced TGF-ß1 activation by cardiac fibroblast contraction and loss of α-SMA expression, which is restored by adding active TGF-ß1. Manipulating integrin protein levels in overexpression and shRNA experiments reveals that both integrins can compensate for each other with respect to TGF-ß1 activation and induction of α-SMA expression. CONCLUSIONS: Integrins αvß5 and αvß3 both control myofibroblast differentiation by activating latent TGF-ß1. Pharmacological targeting of mesenchymal integrins is a possible strategy to selectively block TGF-ß1 activation by cardiac myofibroblasts and progression of fibrosis in the heart.

Asunto(s)

Integrina alfaVbeta3/fisiología , Miofibroblastos/patología , Receptores de Vitronectina/fisiología , Factor de Crecimiento Transformador beta1/fisiología , Actinas/análisis , Adulto , Animales , Diferenciación Celular , Células Cultivadas , Fibrosis , Humanos , Integrina alfaVbeta3/antagonistas & inhibidores , Masculino , Miocardio/patología , Receptores de Vitronectina/antagonistas & inhibidores , Porcinos

RESUMEN

Cooling and heating curves of water confined in partially filled Vycor porous glass were measured for both adsorption and desorption processes. One endothermic and two exothermic peaks were observed for almost all cases. The peak temperature and the enthalpy of the exothermic peak located below 232 K increased initially and then decreased with further increases in the filling factor. These abnormal changes were analyzed based on the liquid-solid transition of nanoconfined water using a core/shell model, and the initial adsorption process of water in this typical mesoporous material with disordered pores is discussed. In addition, an interesting observation is that different peak temperatures for the endothermic peak and an almost constant peak temperature for the exothermic peak were observed at the same filling factor obtained under different sample preparation conditions, that is, adsorption and desorption processes. To compare with the liquid-solid transition temperatures of confined water in fully filled silica-based mesopores of different pore radius, a parameter of the ratio of pore inner surface area to confined liquid volume is proposed in this paper. Referring to this parameter, the core part of confined water in silica-based nanopores has the same liquid-solid transition temperatures. This suggestion is valid for the freezing process of water confined in either fully filled ordered or fully or partially filled disordered pores. For the melting process, different linear changes of melting temperature with the ratio of pore inner surface area to liquid volume were observed for water in disordered and ordered pores.