Ana Luísa Pires

Intermedin elicits a negative inotropic effect in rat papillary muscles mediated by endothelial-derived nitric oxide

Intermedin (IMD) is a novel vasoactive peptide from the calcitonin gene-related peptide (CGRP) implicated in cardiac regulation, yet the contractile effects of IMD remain controversial, since previous studies in vivo and isolated cardiomyocytes documented contradictory results. We hypothesized cardiac endothelial cells involvement in IMD modulation of cardiac function as an explanation for these opposing observations. With this in mind, we investigated the direct action of increasing concentrations of IMD (10(-8) to 10(-6)M) on myocardial performance parameters in rat left ventricular (LV) papillary muscles with and without endocardial endothelium (EE) and in presence of receptor antagonists and intracellular pathways inhibitors. In LV papillary muscles with intact EE, IMD induced a concentration-dependent negative inotropic action (%decrease relative to baseline, at IMD concentration of 10(-6)M, active tension of 14 ± 4%, and maximum velocity of tension rise of 10 ± 4%). These effects were blunted by EE removal, AM receptor antagonist (AM(22-52)), and CGRP receptor antagonist (CGRP(8-37)). Additionally, nitric oxide (NO) synthase inhibition with N(G)-nitro-l-arginine (l-NAME) in muscles with and without EE and guanylyl cyclase inhibition with {1H-[1,2,4]oxadiazole-[4,4-a]-quinoxalin-1-one} not only blunted the negative inotropic action of IMD but also unmasked IMD-positive inotropic effect dependent on CGRP receptor PKA activation. Western blot quantification of phosphorylated cardiac troponin I (P-cTnI) in IMD-treated papillary muscles revealed a significant increase in P-cTnI when compared with untreated muscles, while in l-NAME-pretreated papillary muscles IMD failed to increase P-cTnI. Finally, we found that stimulation of both EE and microvascular endothelial cells with IMD significantly increased NO production by 40 ± 3 and 38 ± 3%, respectively, suggesting the role of cardiac endothelial cells in NO production upon IMD stimulation. Our findings establish IMD negative inotropic effect in isolated myocardium due to NO/cGMP pathway activation with concomitant thin myofilament desensitization by increase in cTnI phosphorylation and provide a coherent explanation for the previously reported contradictory results.

Effects of adrenomedullin on systolic and diastolic myocardial function

Adrenomedullin (AM) effects were studied in rabbit papillary muscles by adding increasing concentrations (10(-10) to 10(-6)M) either alone or after pre-treatment with l-NNA, indomethacin, AM22-52 (AM receptor antagonist), CGRP(8-37) (CGRP receptors antagonist), KT5720 (PKA inhibitor), as well as after endocardial endothelium (EE) removal. Passive length-tension relations were constructed before and after a single concentration of AM (10(-6)M). AM concentration-dependently induced negative inotropic and lusitropic effects, and increased resting muscle length (RL). At 10(-6)M, AT, dT/dt(max) and dT/dt(min) decreased 20.9+/-4.9%, 18.3+/-7.3% and 16.7+/-7.8%, respectively, and RL increased to 1.010+/-0.004L/L(max). Correcting RL to its initial value resulted in a 26.6+/-6.4% decrease of resting tension, indicating decreased muscle stiffness, also patent in the down and rightward shift of the passive length-tension relation. The negative inotropic effect of AM was dependent on its receptor, CGRP receptor, PKA, the EE and NO, while the effects of AM on myocardial stiffness were abolished by EE damage and NO inhibition. This latter effect represents a novel mechanism of acute neurohumoral modulation of diastolic function, suggesting that AM is an important regulator of cardiac filling.

Reverse myocardial effects of intermedin in pressure-overloaded hearts: role of endothelial nitric oxide synthase activity

Intermedin (IMD) is a cardiac endogenous peptide upregulated in several models of heart disease. We show a depressant effect of IMD1−47 on contractility of the normal myocardium, mediated by increased nitric oxide (NO) production due to endothelial nitric oxide synthase (eNOS) phosphorylation. In rat models of cardiac hypertrophy or NO deficiency, IMD1−47 enhances contractility and hastens relaxation associated with phospholamban phosphorylation. This opposing response in normal and diseased myocardium is due to impairment of IMD1−47‐induced eNOS phosphorylation. The results reveal distinct myocardial effects and subcellular mechanisms for IMD1−47 in the hypertrophic heart, suggesting a key role of cardiac endothelial dysfunction with potential pathophysiological relevance.

Highly Active Ruthenium Supported on Magnetically-Recyclable Chitosan-Based Nanocatalyst for Nitroarenes Reduction

A Ru supported on a magnetically separable chitosan‐based nanomaterial (Mn@CS@Ru) was prepared by wet impregnation based on ionic gelation using sodium tripolyphosphate as a cross‐linking agent. The ionic gelation of chitosan leads to a supporting matrix to promote the embedding of manganese(II) ferrite and Ru nanoparticles (NPs) by electrostatic interactions. The effects of the formulation and method parameters on the fabrication process were investigated, and the resulting as‐prepared Mn@CS@Ru nanocatalyst was characterized. The catalytic activity of the Mn@CS@Ru nanomaterial was evaluated in the reduction of 4‐nitrophenol (4‐NP) and 4‐nitroaniline (4‐NA) in the presence of sodium borohydride as a reducing agent at room temperature. The turnover frequency values in the reduction of 4‐NP and 4‐NA were 273.9 and 336.5 min−1, respectively, which were attributed to the very small size of the hybrid nanomaterial (32.0±2.8 nm with 3.9±0.1 nm Ru NPs) that provided a large surface‐area‐to‐volume ratio for the chemical reaction. Furthermore, the hybrid nanocatalyst was recovered easily by magnetic separation after the catalytic reaction and could be reused in at least 10 cycles without a loss of catalytic activity, which confirms its high stability. The present route is a new approach to synthesize highly active magnetic heterogeneous catalysts for the reduction of nitroarenes based on metallic NPs with easy accessibility, excellent activity, and convenient recovery.