Avaliação dos efeitos induzidos pelo 2-Nitrato-1, 3-Dibutoxipropano (NDBP) sobre o sistema cardiovascular de ratos normotensos - abordagens en vivo e in vitro

Abstract

Organic nitrates are nitric oxide (NO) donors used in the treatment of cardiovascular diseases mimicking the role of endogenous NO. This study evaluated organic nitrates newly synthesized from glycerin, which cardiovascular actions had not yet been investigated. Therefore, the cardiovascular effects produced by organic nitrates derived from glycerin: 2-nitrate-1,3-dimethoxypropan (NDMP), 2-nitrate-1,3-diethoxypropan (NDEP), 2-nitrate-1 ,3-dipropoxypropan (NDPP) and 2-nitrate-1,3-dibutoxypropan (NDBP) in rats were investigated using in vitro and in vivo approaches. For in vitro studies, animals were euthanized and the superior mesenteric artery was isolated. Artery rings were kept in tanks with Tyrode at 37 ° C aerated with carbogen, then were attached to a force transducer (Fort 10, WPI, Sarasota, USA) coupled to a data acquisition system data (Miobath-4, WPI, Sarasota, USA) under a tension of 0.75 g for 1 hour. After this period, preparations were pre-contracted with phenylephrine (FEN) 10 μM or KCl 80 mM and then increasing concentrations of organic nitrates were cumulatively added. The nitrate with the most promising effects was selected for further studies and concentration-response curves of the compound selected in the presence of HDX, a hijacker of NO; ODQ, inhibitor of soluble guanylyl cyclase, KCl 20 mM, a modulator of potassium efflux, and blockers for calcium-sensitive potassium channel (TEA, 1 mM), blockers for ATP-sensitive potassium channel (GLIB, 1 M) and blockers for voltage-operated potassium channel (4-AP, 1 mM) were obtained. All compounds showed vasorelaxant activity endothelium-independent in superior mesenteric artery rings, being the NDBP was the most potent agent with Emax = 105.4 ± 2.7% in rings pre-contracted with FEN and Emax = 82, 7 ± 7.9% in KCl 80 mM induced contraction. The vasorelaxation was significantly attenuated in the presence of HDX and ODQ with Emax = 62.8 ± 14.9% and Emax = 15.2 ± 9.2%, respectively. In the presence of KCl 20 mM the vasorelaxat response was also reduced [Emax = 70.6 ± 15.02%] as well as in the presence of TEA [Emax = 87.97 ± 5.78%]; GLIB [Emax = 78, 2 ± 6.5%] and 4-AP, a lesser extent [Emax = 94.65 ± 6.6%]. For in vivo studies, we investigated the changes in blood pressure (BP) and heart rate (HR) in conscious rats treated acutely with NDBP. Intravenous administration of NDBP (2, 5, 10, 15 and 20 mg/kg, randomly) produced hypotension (-6 ± 1.7, -22 ± 6.8, -58 ± 3.7, -70 ± 5.5, -77 ± 5.4 mmHg) and bradycardia (-12 ± 5, -40 ± 19.7, -133 ± 18.6, -179 ± 23.5, and -266 ± 12.4 bpm) in a dose-dependent manner. Thus, the vasorrelaxant response produced by NDBP possibly involves the NO release and subsequent activation of the CGs/GMPc/PKG pathway and BKCa, KATP and KV channels. These mechanism of action may be contributing to hypotension and bradycardia showed in non-anesthetized normotensive rats.