Fenofibrate and Pioglitazone Do Not Ameliorate the Altered Vascular Reactivity in Aorta of Isoproterenol-treated Rats

Chronic stimulation of beta-adrenoceptors with isoproterenol induces alteration of vascular reactivity and increases local proinflammatory cytokines. We investigated whether fenofibrate and pioglitazone, PPAR-alpha and -gamma agonists, respectively, improve the changes in vascular reactivity induced by isoproterenol. Wistar rats received isoproterenol (0.3 mg.kg(-1).day(-1), SC) or vehicle (CT) plus fenofibrate (alpha, 100 mg.kg(-1).day(-1), PO), pioglitazone (gamma, 2.5 mg.kg(-1).day(-1), PO), or water for 7 days. In aortas, isoproterenol treatment enhanced the maximal response (Rmax) to phenylephrine (10(-10) to 10(-4) M) compared to CT as previously demonstrated. The effects of endothelium removal (E-) or L-NAME incubation (100 mu M) on the phenylephrine response were smaller in isoproterenol-treated animals compared to CT while superoxide dismutase (SOD, 150 U/mL) significantly reduced the Rmax to phenylephrine to CT levels. Neither fenofibrate nor pioglitazone changed the effects induced by isoproterenol in aorta. E-, L-NAME, or SOD effects were similar between CT alpha and CT. However, pioglitazone per se increased Rmax to phenylephrine (CT: 59 +/- 4 versus CT gamma: 72 +/- 5 % of contraction to KCl). E- or L-NAME effects were reduced in CT gamma compared to CT, and SOD normalized the altered reactivity to phenylephrine in the CT gamma group. In conclusion, neither fenofibrate nor pioglitazone ameliorates the altered vascular reactivity present in aorta from isoproterenol-treated rats. Moreover, pioglitazone per se induced endothelial dysfunction and increased phenylephrine-induced contraction in aorta.

Time-dependent increases in ouabain-sensitive Na(+), K(+)-ATPase activity in aortas from diabetic rats: The role of prostanoids and protein kinase C

Aims: Na(+), K(+)-ATPase activity contributes to the regulation of vascular contractility and it has been suggested that vascular Na(+), K(+)-ATPase activity may be altered during the progression of diabetes; however the mechanisms involved in the altered Na(+), K(+)-ATPase activity changes remain unclear. Thus, the aim of the present study was to evaluate ouabain-sensitive Na(+), K(+)-ATPase activity and the mechanism(s) responsible for any alterations on this activity in aortas from 1- and 4-week streptozotocin-pretreated (50 mg kg(-1), i.v.) rats. Main methods: Aortic rings were used to evaluate the relaxation induced by KCl (1-10 mM) in the presence and absence of ouabain (0.1 mmol/L) as an index of ouabain-sensitive Na(+), K(+)-ATPase activity. Protein expression of COX-2 and p-PKC-beta II in aortas were also investigated. Key findings: Ouabain-sensitive Na(+), K(+)-ATPase activity was unaltered following 1-week of streptozotocin administration, but was increased in the 4-week diabetic aorta (27%). Endothelium removal or nitric oxide synthase inhibition with L-NAME decreased ouabain-sensitive Na(+), K(+)-ATPase activity only in control aortas. In denuded aortic rings, indomethacin. NS-398, ridogrel or Go-6976 normalized ouabain-sensitive Na(+), K(+)-ATPase activity in 4-week diabetic rats. In addition, COX-2 (51%) and p-PKC-beta II (59%) protein expression were increased in 4-week diabetic aortas compared to controls. Significance: In conclusion, diabetes led to a time-dependent increase in ouabain-sensitive Na(+), K(+)-ATPase activity. The main mechanism involved in this activation is the release of TxA(2)/PGH(2) by COX-2 in smooth muscle cells, linked to activation of the PKC pathway. (C) 2010 Elsevier Inc. All rights reserved.

Clopidogrel, independent of the vascular P2Y(12) receptor, improves arterial function in small mesenteric arteries from Angll-hypertensive rats

The P2Y(12) receptor antagonist clopidogrel blocks platelet aggregation, improves systemic endothelial nitric oxide bioavailability and has anti-inflammatory effects. Since P2Y(12) receptors have been identified in the vasculature, we hypothesized that clopidogrel ameliorates Angll (angiotensin II)-induced vascular functional changes by blockade of P2Y(12) receptors in the vasculature. Male Sprague Dawley rats were infused with Angll (60 ng/min) or vehicle for 14 days. The animals were treated with clopidogrel (10 mg . kg(-1) of body weight . day(-1)) or vehicle. Vascular reactivity was evaluated in second-order mesenteric arteries. Clopidogrel treatment did not change systolic blood pressure [(mmHg) control-vehicle, 117 +/- 7.1 versus control-clopidogrel, 125 +/- 4.2; Angll vehicle, 197 +/- 10.7 versus Angll clopidogrel, 198 +/- 5.2], but it normalized increased phenylephrine-induced vascular contractions [(%KCI) vehicle-treated, 182.2 +/- 18% versus clopidogrel, 133 +/- 14%), as well as impaired vasodilation to acetylcholine [(%) vehicle-treated, 71.7 +/- 2.2 versus clopidogrel, 85.3 +/- 2.8) in Angll-treated animals. Vascular expression of P2Y(12) receptor was determined by Western blot. Pharmacological characterization of vascular P2Y(12) was performed with the P2Y(12) agonist 2-MeS-ADP [2-(methylthio) adenosine 5`-trihydrogen diphosphate trisodium]. Although 2-MeS-ADP induced endothelium-dependent relaxation [(Emax %) = 71 +/- 12%) as well as contractile vascular responses (Emax % = 83 +/- 12%), these actions are not mediated by P2Y(12) receptor activation. 2-MeS-ADP produced similar vascular responses in control and Angll rats. These results indicate potential effects of clopidogrel, such as improvement of hypertension-related vascular functional changes that are not associated with direct actions of clopidogrel in the vasculature, supporting the concept that activated platelets contribute to endothelial dysfunction, possibly via impaired nitric oxide bioavailability.

Functionalization of PAMAM dendrimers with [Ru-III(edta)(H2O)](-)

The anchoring of K[Ru-III(edta)(Cl)] on poly(amidoamine) dendrimers (PAMAM of three generations G(x)/Ru (x = 0, 2 and 3)) through a peptide type bond yielded the aquo species, [Ru-III(edta)(H2O)] on dendrimer surface, and upon NO exposure, yielded their nitrosyl analogues, Gx/RuNO. Characterization of these compounds by elemental analysis, and a UV-vis, IR and C-13 NMR spectroscopies indicated the immobilization of 4,12 and 29 molecules of [Ru-III(edta)(H2O)](-) or of the nitrosyl complex [Ru(II)edta)NO] on the dendrimer surface for G(X) = 0, 2 and 3, respectively. For each complex the electrochemical spectrum presented only one redox process with redox potential values of -0.20 and -0.32 V(vs SCE) attributed to the Ru/Run and NO+/NO0 couples in G(x)/Ru and G./RuNO, respectively. The one-electron reduction of Gx/RuNO` generates Gx/RuNOo, which undergoes aquation with a k(-NO) of 2.1 +/- 0.7 x 10(-3) s(-1) (pH 1.0, mu = 0.2 mol/L, CF3COOH/NaCF3COO, 25 degrees C). The Gx/RuNO species induced a relaxing effect in aortic rings denuded of endothelium and exhibited in vitro assay trypanocidal activity. (c) 2008 Elsevier Inc. All rights reserved.

Níveis de óxido nítrico e fibrinogênico em pacientes com síndrome dos ovários policísticos : assocação com medidas antropométricas, perfil hormonal e metabólico

Introdução: O óxido nítrico (NO) e o fibrinogênio são importantes marcadores de disfunção endotelial e de disfibrinólise, respectivamente, sendo essas alterações relacionadas à resistência insulínica. A Síndrome dos Ovários Policísticos (PCOS), caracterizada por irregularidade menstrual e anovulação crônica, está associada com resistência à insulina e hiperinsulinemia em porcentagem considerável dos casos. A disfunção endotelial, decorrente da resistência insulínica e de alterações nos níveis de NO e de fibrinogênio, poderia conferir a essas pacientes um maior risco de doença macrovascular. Objetivo: Avaliar níveis séricos de óxido nítrico e de fibrinogênio em pacientes com PCOS e compará-los aos de pacientes com Hirsutismo Idiopático (HI). Materiais e métodos: Estudo transversal. Foram avaliados níveis de óxido nítrico e de fibrinogênio e suas associações com variáveis antropométricas, metabólicas e hormonais em 26 pacientes hirsutas com PCOS e 20 pacientes do grupo controle com HI (ciclos regulares e ovulatórios, níveis normais de androgênios e hirsutismo isolado). Resultados: Os grupos foram semelhantes quanto à história familiar de diabete e gravidade do hirsutismo pelo escore de Ferriman-Gallway. Não houve diferenças significativas nos níveis de NO e de fibrinogênio entre os grupos PCOS e HI. Entretanto, nas pacientes com PCOS, insulina e HOMA (Homeostatic Model Assessment) foram negativamente correlacionadas com níveis de NO (r=-0.42 p<0.03).Considerando-se todas as pacientes, idade, índice de massa corporal (IMC) e circunferência da cintura tiveram correlação positiva com níveis de fibrinogênio. Conclusão: Os resultados mostram associação negativa e não dependente do IMC entre NO e resistência insulínica, mas não com níveis de androgênios, apenas nas pacientes com PCOS. Assim, as estratégias clínicas que objetivam a redução da resistência insulínica podem trazer benefícios às pacientes com PCOS não somente para a prevenção de diabetes e de dislipidemia, mas também para a redução do risco de disfunção endotelial nessas pacientes.

Oxidative stress and inflammatory mediators contribute to endothelial dysfunction in high-fat diet-induced obesity in mice

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Differential vascular adaptive response to stress exposure in male and female rats: Role of gonadal hormones and endothelial cells

Although there are reports concerning a vascular adaptive response to stress in males, this is not yet defined in females. The aim of this study was to delineate functional gender differences in the rat vascular adaptive response to stress and to determine the ability of sex hormones to modulate the stress-induced vascular adaptive response. Responses to noradrenaline were evaluated in aortas, with and without endothelium, from intact, gonadectomized and gonadectomized-hormone-replaced males and females submitted or not to stress (2-h immobilization). Reactivity of the aorta of stressed and non-stressed intact males and females (n = 6-14 per group) was also examined in the presence of L-NAME or indomethacin. Stress decreased and gonadectomy increased maximal responses to noradrenaline in aortas with intact endothelium from both genders. Stress also reduced noradrenaline potency in males. In females, but not males, stress decreased the gonadectomy-induced noradrenaline hyper-reactivity to near that of intact non-stressed rats. Hormone replacement restored the gonadectomy-induced impaired vascular adaptive response to stress. L-NAME, but not indomethacin, abolished the stress-induced decrease in aorta reactivity of males and females. None of the procedures altered reactivity of aortas denuded of endothelium. Conclusion: Stress-induced vascular adaptive responses show gender differences. The magnitude of the adaptive response is dependent on testicular hormones and involves endothelial nitric oxide-system hyperactivity.

Alterações vasculares em ratos obesos por dieta rica em gordura: papel da Via L-arginina/NO Endotelial

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Reactivity of mesenteric and aortic rings from trained rats fed with high caloric diet

A strong association between the benefits of physical exercise on the cardiovascular disease with an improvement of the endothelium-derived relaxing factor production has been consistently shown. The purpose of this study was to evaluate the effect of exercise training associated with high caloric diet in the reactivity of rat mesenteric and aortic rings. Experimental protocol consisted of 4 weeks of high caloric diet consumption previous to 4 weeks of run training (1.2 km/h, 0% grade, in sessions of 60 min, 5 days/week). Concentrations of triglycerides, glucose, insulin and nitrite/nitrate levels were measured and atherogenic index was calculated. Concentration-response curves to acetylcholine (10 nM-100 mu M), sodium nitroprusside (100 pM-100 nM) and phenylephrine (1 nM-3 mu M) were obtained. Exercise training reduced body mass (6%) and triglyceride levels (about 54%), without changes in glucose and insulin concentrations. An improvement of endothelium-dependent relaxation responses to acetylcholine in mesenteric and aortic rings was observed in trained group. No changes were seen for sodium nitroprusside and phenylephrine. In conclusion, our study is the first to show clearly that run training promotes an improvement of the endothelium-dependent relaxing response in aorta and mesenteric rings from rats fed with high caloric diet and that is associated with increase of NO production. (c) 2006 Elsevier B.V. All rights reserved.

High-fat diet associated with obesity induces impairment of mouse corpus cavernosum responses

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Influence of acute pancreatitis on the in vitro responsiveness of rat mesenteric and pulmonary arteries

Background: Acute pancreatitis is an inflammatory disease characterized by local tissue injury and systemic inflammatory response leading to massive nitric oxide (NO) production and haemodynamic disturbances. Therefore, the aim of this work was to evaluate the vascular reactivity of pulmonary and mesenteric artery rings from rats submitted to experimental pancreatitis.Male Wistar rats were divided into three groups: saline (SAL); tauracholate (TAU) and phospholipase A(2) (PLA(2)). Pancreatitis was induced by administration of TAU or PLA(2) from Naja mocambique mocambique into the common bile duct of rats, and after 4 h of duct injection the animals were sacrificed. Concentration-response curves to acetylcholine (ACh), sodium nitroprusside (SNP) and phenylephrine (PHE) in isolated mesenteric and pulmonary arteries were obtained. Potency (pEC(50)) and maximal responses (E(MAX)) were determined. Blood samples were collected for biochemical analysis.Results: In mesenteric rings, the potency for ACh was significantly decreased from animals treated with TAU (about 4.2-fold) or PLA(2) (about 6.9-fold) compared to saline group without changes in the maximal responses. Neither pEC(50) nor E(MAX) values for Ach were altered in pulmonary rings in any group. Similarly, the pEC(50) and the E(MAX) values for SNP were not changed in both preparations in any group. The potency for PHE was significantly decreased in rat mesenteric and pulmonary rings from TAU group compared to SAL group (about 2.2- and 2.69-fold, for mesenteric and pulmonary rings, respectively). No changes were seen in the E(MAX) for PHE. The nitrite/nitrate (NO(x)(-)) levels were markedly increased in animals submitted to acute pancreatitis as compared to SAL group, approximately 76 and 68% in TAU and PLA(2) protocol, respectively.Conclusion: Acute pancreatitis provoked deleterious effects in endothelium-dependent relaxing response for ACh in mesenteric rings that were strongly associated with high plasma NO(x)(-) levels as consequence of intense inflammatory responses. Furthermore, the subsensitivity of contractile response to PHE in both mesenteric and pulmonary rings might be due to the complications of this pathological condition in the early stage of pancreatitis.

Interaction between Advanced Glycation End Products Formation and Vascular Responses in Femoral and Coronary Arteries from Exercised Diabetic Rats

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Improvement in relaxation response in corpus cavernosum from trained rats

Objectives. To evaluate the contractile and relaxing responses in rat corpus cavernosum (RCC) from rats after 8 weeks of run training, because erectile function is highly dependent on nitric oxide (NO) from nitrergic fibers or endothelium. Physical activity enhances NO production and improves endothelial function, with beneficial effects on cardiovascular disease.Methods. The training program consisted of 8 weeks of run training, 5 days/wk, and each session lasted 60 minutes. The RCC was isolated, and concentration-response curves to NO, acetylcholine, sodium nitroprusside, phenylephrine, and endothelin were obtained. The excitatory and inhibitory effects of electrical field stimulation (2 to 32 Hz) were also evaluated.Results. NO (0.1 to 100 muM) and sodium nitroprusside (0.01 to 1000 muM) produced a relaxing effect in RCC in a dose-dependent manner, with the maximal responses to NO (control 62% +/- 4%, trained 88% +/- 3%) and sodium nitroprusside (control 83% +/- 3%, trained 95% +/- 2%) significantly enhanced after 8 weeks of run training. However, acetylcholine-induced relaxations were not affected by exercise. Similarly, electrical field stimulation-induced relaxations were significantly increased in RCC from trained rats at 2 Hz (control 2.4% +/- 0.3%, trained 4.2% +/- 0.5%) and 4 Hz (control 5.3% +/- 1.2%, trained 12.5% +/- 1.7%). The contractile sensitivity of RCC to phenylephrine (0.01 to 100 AM) and endothelin (0.01 to 100 nM) was not modified by training exercise.Conclusions. Our findings suggest that run training enhances functional responses in rat RCC that involves increases in the NO-cyclic guanosine monophosphate signaling pathway by endothelium-independent mechanisms that is not accompanied by changes in contractile sensitivity. (C) 2004 Elsevier B.V.

Atypical beta-adrenoceptor subtypes mediate relaxations of rabbit corpus cavernosum

This study was performed to characterize the beta-adrenoceptor population in rabbit isolated corpus cavernosum (RbCC) by using nonselective and selective beta-adrenoceptor agonists and antagonists in functional assays. Metaproterenol, ritodrine, fenoterol, and 8-hydroxy-5-[(1R)-1-hydroxy-2-[N-[(1R)-2-(rho-methoxy-phenyl)1-methylethyl] amino] ethyl] carbostyril (TA 2005) (3-100 nmol each) dose dependently relaxed the RbCC preparations. These relaxations were markedly reduced by N-omega-nitro-L-arginine methyl ester (L-NAME; 10 muM) and 1H-[1,2,4]-oxadiazolo-[4,3,-a]quinoxalin-1-one (ODQ) (10 muM), whereas the adenylyl cyclase inhibitor SQ 22,536 [9-(2-tetrahydrofuryl)adenine] (10 muM) had no effect. In contrast, neither L-NAME nor ODQ affected the isoproterenol-induced RbCC relaxations, but SQ 22,536 abolished this response. Sildenafil (1 muM) significantly potentiated the relaxations induced by beta(2)-agonists without affecting the isoproterenol-evoked relaxations. Rolipram (10 muM) enhanced the relaxations elicited by isoproterenol but had no effect on those induced by the selective beta(2) agonists. Propranolol and (+/-)-1-[2,3-(dihydro-7-methyl-1H-inden-4-yl)oxy]-3-[(1-methylethyl)amino]-2-butanolhydrochloride (ICI 118,551) determined a rightward shift in the concentration-response curves to isoproterenol in a noncompetitive manner with a reduction of maximum response at the highest antagonist concentration, with the slope values significantly different from unity. Propranolol and ICI 118,551 had no effect on the relaxations elicited by fenoterol, TA 2005, metaproterenol, and ritodrine. Atenolol and 1-[2-((3-carbamoyl-4-hydroxy)phenoxy) ethylamino]-3-[4-(1-methyl-4-trifluoromethyl-2-imidazolyl)-phenoxy]2-propanol methanesulfonate (CGP 20712A) (0.1-10 muM) failed to affect the relaxations induced by all tested beta-adrenoceptor agonists. Our study revealed the existence of two atypical beta-adrenoceptors in the rabbit erectile tissue. Isoproterenol relaxes the rabbit cavernosal tissue by activating atypical beta-adrenoceptors coupled to adenylyl cyclase pathway, whereas the selective beta2-adrenoceptor agonists relax the RbCC tissue through another atypical beta-adrenoceptor subtype coupled to nitric oxide release from the sinusoidal endothelium.

Papel da regiao AV3V no choque septico experimental em ratos tratados com NaCl 7,5%

The effect of intravenous infusion of hypertonic saline (HS) on the recovery of mean arterial pressure (MAP) during septic shock was studied in sham-operated rats and in rats with electrolytic lesion in the anteroventral third ventricle (AV3V) region. Our results show that intravenous HS infusion in rats treated with endotoxin (Etx) partially restores MAP, but when we have a severe shock produced by Etx, HS was not able to reverse the hypotension. We also show that the integrity of the AV3V region is essential for the protective action of HS in endotoxin shock. It is possible that NO production contributes to the deleterious effect of endotoxin. So, the unraveling of the release of NO by the vascular endothelium and their role as regulators of vascular tone is increasing our understanding of the physiology and pathophysiology of the cardiovascular system and will therefore enhance the possibilities of preventing and treating endotoxin shock.