Long-term nitric oxide inhibition and chronotropic responses in rat isolated right atria

The long-term administration of nitric oxide synthesis inhibitors induces arterial hypertension accompanied by left ventricular hypertrophy and myocardial ischemic lesions. Because the enhancement of sympathetic drive has been implicated in these phenomena, the current study was performed to determine the potency of β-adrenoceptor agonists and muscarinic agonists on the spontaneous rate of isolated right atria from rats given long-term treatment with the nitric oxide inhibitor N(ω)-nitro-L-arginine methyl ester (L-NAME). Atrial lesions induced by long-term treatment with L-NAME were also evaluated. Long-term L-NAME treatment caused a time-dependent, significant (P<0.05) increase in tail-cuff pressure compared with control animals. Our results showed that the potency of isoproterenol, norepinephrine, carbachol, and pilocarpine in isolated right atria from rats given long-term treatment with L-NAME for 7, 15, 30, and 60 days was not affected as compared with control animals. Addition of L-NAME in vitro (100 μmol/L) affected neither basal rate nor chronotropic response for isoproterenol and norepinephrine in rat heart. Stereological analysis of the right atria at 15 and 30 days revealed a significant increase on amount of fibrous tissues in L-NAME- treated groups (27±2.3% and 28±1.3% for 15 and 30 days, respectively; P<0.05) as compared with the control group (22±1.1%). Our results indicate that nitric oxide does not to interfere with β-adrenoceptor-mediated and muscarinic receptor-mediated chronotropic responses.

Influence of arginine vasopressin receptor and nitric oxide on the water, sodium intake and arterial blood pressure induce by angiotensin injected into third ventricle of the brain

As several structures of the central nervous system are involved in the control of hydromineral and cardiovascular balance we investigated whether the natriorhexigenic and pressor response induced by the injection of ANG II into the 3rd V could be mediated by vasopressinergic and nitrergic system. Male Holtzman rats weighing 200-250 g with cannulae implanted into the 3rd V were used. The drugs were injected in 0.5 μL over 30-60 sec. Controls were injected with a similar volume of 0.15 M NaCl. ANGII increased the water intake vs control. AVPA injected into 3rd V prior to ANGII decreased the dipsogenic effect of ANGII. L-arginine also decreased the water intake induced by ANGII. AVPA plus L-arginine inhibit the water intake induced by ANGII. 7NIT injected prior to ANGII potentiated the dipsogenic effect of ANGII. Pre-treatment with ANGII increased the sodium ingestion vs control. AVPA decreased the ANGII effect in sodium intake. L-arginine also decreased the natriorhexigenic effect of ANGII. The combination of L-arginine and AVPA inhibit the sodium intake induced by ANGII. 7NIT injected prior to ANGII potentiated the sodium intake induced by ANGII. ANGII induced an increase in Mean Arterial Pressure (MAP) vs control. AVPA and L-arginine induced a decreased in the pressor effect of ANGII. The combination of L-arginine and AVPA inhibit the pressor effect of ANGII. 7NIT injected prior to ANGII into 3rd V potentiated the pressor effect of ANGII. These data suggest that arginine vasopressin V 1 receptors and Nitric Oxide (NO) within the circumventricular structures may be involved in sodium intake and pressor response induced by the activation of ANGII receptors within the circumventricular neurons. These studies revealed the involvement of sodium appetite by utilizing the angiotensinergic, vasopressinergic and nitrergic system in the central regulation of blood pressure. © 2006 Asian Network for Scientific Information.

Nitrergic pathways and L-type calcium channel of MnPO influencing cardiovascular homeostasis

The median preoptic nucleus (MnPO) is one of most important site of the lamina terminalis implicated in the regulation of hydro electrolytic and cardiovascular balance. The purpose of this study was to determine the effect of L-Type calcium channel antagonist, nifedipine, on the increase of median arterial blood pressure (MAP) induce by angiotensin II (ANG II) injected into the MnPO. The influence of nitric oxide (NO) on nifedipine antipressor action has also been studied by utilizing N W-nitro-L-arginine methyl ester (L-NAME) (40 μg 0.2 μL -1) a NO synthase inhibitor (NOSI), 7-nitroindazole (7-NIT) (40 μg 0.2 μL -1), a specific neuronal NO synthase inhibitor (nNOSI) and sodium nitroprusside (SNP) (20 μg 0.2 μL -1) a NO donor agent. We have also investigated the central role of losartan and PD123349 (20 nmol 0.2 μL -1), AT 1 and AT 2, respectively (selective non peptide ANG II receptor antagonists), in the pressor effect of ANG II (25 pmol 0.2 μL -1) injected into the MnPO. Male Wistar rats weighting 200-250 g, with cannulae implanted into the MnPO were utilized. Losartan injected into the MnPO, prior to ANG II, blocked the pressor effect of ANGII. PD 123319 only decreased the pressor effect of ANG II. Rats pre-treated with either 50 μg 0.2 μL -1 or 100 μg 0.2 μL -1 of nifedipine, followed by 25 pmol 0.2 μL -1 of ANG II, decreased ANG II-pressor effect. L-NAME potentiated the pressor effect of ANG II. 7-NIT injected prior to ANG II into the MnPO also potentiated the pressor effect of ANGII but with less intensity than that of L-NAME. SNP injected prior to ANG II blocked the pressor effect of ANG II. The potentiation action of L-NAME and 7-NIT on ANG II-pressor effect was blocked by prior injection of nifedipine. The results described in this study provide evidence that calcium channels play important roles in central ANG II-induced pressor effect. The structures containing NO in the brain, such as MnPO, include both endothelial and neuronal cells, which might be responsible for the influence of nifedipine on the pressor effect of ANG II. These data have shown the functional relationship between L-Type calcium channel and a free radical gas NO in the MnPO, on the control of ANG II-induced pressor effect acting in AT 1 and AT 2 receptors.

Effects of nitric oxide and arginine vasopressin on sodium intake induced by central angiotensin II. Part 2

We study the effects of angiotensin receptors antagonists, arginine vasopressin receptor antagonist, L-arginine and L-NAME, injected into supraoptic nucleus of the hypothalamus (SON) on sodium intake induced by the injection of angiotensin II (ANGII). Holtzman rats weighing 200-250 g with canulae implanted into the SON were used. The drugs were injected in 0.5 μL over 30-60 sec. Sodium intake after injection of saline SAL+SAL 0.15 M NaCl was 0.10±00.1 mL 2 h -1; SAL+ANGII injected into SON increased sodium intake. Losartan injected prior to ANGII into SON decreased sodium intake induced by ANGII. PD123319 injected prior to ANGII produced no changes in sodium intake induced by ANGII. AVPA receptor V 1 antagonist injected prior to ANGII reduced sodium intake with a less intensity than losartan. L-arginine injected prior to ANGII decreases sodium intake at a same intensity than losartan. L-NAME injected prior to ANGII potentiated sodium intake induced by ANGII. Losartan injected simultaneously with L-arginine prior to ANGII blocked the natriorexigenic effect of ANGII. These results confirm the importance of SON in the control of sodium intake. Also suggest that both AT 1 and arginine vasopressin V 1 receptors interact with nitrergic pathways within the SON influencing the sodium metabolism by changing sodium appetite induced by ANGII. © 2007 Asian Network for Scientific Information.

Effects of nitric oxide and arginine vasopressin on water intake induced by central angiotensin II. Part 1

We determined the effects of AT 1 and AT 2 (selective no peptides antagonists angiotensin receptors), arginine vasopressin V 1 receptor antagonist as well as L-arginine, a nitric oxide donor and N W-nitro-L-arginine methyl ester (L-NAME), a nitric oxide synthase inhibitor, injected into supraoptic nucleus (SON) on water and sodium intake induced by the injection of angiotensin II (ANGII). Male Holtzman rats weighing 200-250 g with canulae implanted into the SON were used. The drugs were injected in 0.5 μL over 30-60 sec. The water intake after injection of saline SAL+SAL 0.15 M NaCl was 0.40±0.1 mL 2 h -1; SAL+ANGII increase water intake. Losartan decreased the water intake induced by ANGII. PD123319 injected prior to produce no change in water intake induced by ANGII. AVPA prior to ANGII reduced the water intake with a less intensity than losartan. L-arginine prior to ANGII decreases the water intake at a same intensity than losartan. L-NAME prior to ANGII potentiated the dipsogenic effect of ANGII. Losartan injected simultaneously with L-arginine prior to ANGII blocked the dipsogenic effect of ANGII. These results confirm the importance of SON in the control of water intake and strongly suggest that AT 1, V 1 receptors interact with nitrergic pathways within the SON influencing the dipsogenic effect of ANGII.

Efeitos terapêuticos da suplementação de L-arginina nas doenças cardiovasculares e endócrino-metabólicas

Nitric oxide is synthesized from L-arginine and plays an important role in blood pressure regulation, platelets aggregation and atherosclerosis development. Most of the studies have shown that the beneficial effects of the L-arginine supplementation are related to an increasing of nitric oxide bioavailability to the cells, improving the endothelial dysfunction, decreasing oxidative stress, ameliorating lipid profile and insulin resistance. However some studies show conflicting results. Considering the role of the endothelium on the pathogenesis of the cardiovascular diseases as well as on the endocrine-metabolic diseases, this review will update studies involving the role of nitric oxide and its signaling pathways in the regulation of vascular function. Furthermore, this review will focus on the main results of the clinical trials using oral L-arginine supplementation, with or without physical exercise, in an attempt to obtain beneficial effects on the cardiovascular and endocrine-metabolic systems in patients and healthy subjects.

Efeito da suplementação com L-Arginina sobre a produção de mediadores inflamatórios e fatores regenerativos pelo tecido muscular esquelético decorrente de uma sessão de exercício resistido de alta intensidade

Pós-graduação em Ciências Fisiológicas - FOA

Efeitos do treinamento físico associado à suplementação de L-arginina na resposta relaxante de artéria femoral de ratos

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Uso da L-arginina nos processos de capacitação espermática e fecundação in vitro de oócitos bovinos

O objetivo deste trabalho foi avaliar o uso da L-arginina nos processos de capacitação espermática e fecundação in vitro (FIV), analisando sua influência no desenvolvimento embrionário, utilizando o sêmen de dois touros (Bos taurus e Bos indicus). No experimento 1, os espermatozóides foram incubados, sem a presença de oócitos, durante 0, 1, 2 e 3 h em meio de FIV adicionado de 0, 1, 10 e 50 mM de L-arginina, sendo analisada a taxa de reação acrossômica. No experimento 2, espermatozóides e oócitos foram incubados em meio de FIV acrescido com as concentrações de L-arginina citadas anteriormente, durante aproximadamente 30 h. Os oócitos bovinos foram maturados in vitro (MIV) e o subsequente cultivo embrionário (CIV) foi realizado sobre monocamada de células da granulosa, em meio SOF, sendo avaliadas as taxas de fecundação (18 hpi), clivagem e blastocisto (2º e 7º dia de cultivo, respectivamente). A dosagem de NO₃ ^-/NO₂ ^- produzido durante a FIV foi realizada através do método colorimétrico de Griess. Para análise estatística dos dados, foi utilizado a ANOVA, com nível de significância de 5%. A Larginina (1 mM), quando adicionada ao meio de capacitação espermática, durante duas horas, aumentou a taxa de reação acrossômica em relação ao controle (31,1±2,78 vs 23,4±2,65) em Bos taurus. A adição de L-arginina (50 mM) ao meio de FIV (experimento 2), tanto em Bos taurus quanto em Bos indicus, diminuiu as taxas de clivagem (78,7±2,17 vs 65,7±9,32; 72,7±3,36 vs 45±7,12; respectivamente) e blastocisto (39,4±3,78 vs 15,2±6,12; 39,4±4,39 vs 16±8,54; respectivamente) em relação ao controle. Sendo assim, observou-se que a L-arginina aumentou a taxa de reação acrossômica em Bos taurus, porém reduziu as taxas de clivagem e blastocistos em ambos os touros, sem influenciar na qualidade do embrião.

A intensificação do comportamento tipo ansiedade induzido por cafeína em Daniorerio(zebrafish) é prevenida pelo tratamento com α-Tocoferol e L-NAME

O crescente consumo de bebidas com elevado teor de cafeína pode resultar no aparecimento de sintomas provenientes do transtorno de ansiedade induzida por essa droga. Atualmente, tem-se utilizado a cafeína como um indutor farmacológico do comportamento tipo ansiedade e essa indução pode facilitar a melhor compreensão da relação entre alterações comportamentais e os mecanismos de ação envolvidos nesse efeito, portanto o presente trabalho propôs que a via nitrérgica poderia ser um mecanismo chave para explicar os efeitos comportamentais produzidos pela cafeína e que esses efeitos poderiam ser revertidos por um antioxidante, logo, no presente trabalho nós tivemos como objetivo avaliar o possível efeito do L-NAME e do α-tocoferol no comportamento tipo ansiedade ampliado pela cafeína nos testes de preferência claro/escuro (PCE) e distribuição vertical eliciada pela novidade (DVN) em Daniorerio. Foram utilizados peixes da espécie Daniorerio(n=178) subdivididos nos seguintes grupos experimentais: SAL – salina 0,9%; CAF – cafeína 100 mg/kg; DMSO – dimetilsulfóxido 0,1%; L-NAME - (N -Nitro-L-arginina-metil éster hidrocloreto) 10 mg/kg; TF – α-tocoferol 1 mg/kg (receberam apenas uma injeção por i.p); SAL + SAL; DMSO + SAL; SAL + CAF; L-NAME + SAL; L-NAME +CAF; TF + CAF (receberam duas injeções seguidas, uma injeção de cada substância na forma de cotratamento, por i.p). Os animais foram submetidos ao teste de preferência claro/escuro e de distribuição vertical eliciada pela novidade. Todos os testes foram filmados e os vídeos foram avaliados utilizando o X-PLO-RAT. Os dados foram expressos em média ± erro padrão. Foi aplicado o teste de normalidade utilizando o teste Shapiro-Wilk e o teste paramétrico ANOVA de uma via com pós-teste Tukey, considerando significativos valores com p<0,05. Nós demonstramos que o α-tocoferol na dose de 1 mg/kg reverteu todos os parâmetros do comportamento tipo ansiedade ampliado pela cafeína nos testes de PCE e do DVN e esse efeito foi semelhante ao observado quando administrado um inibidor da enzima óxido nítrico sintase (NOS), L-NAME. Portanto, o presente trabalho demonstrou pela primeira vez que o efeito comportamental ampliado pela cafeína no teste escotáxico e no DVN pode ser modulado pelo sistema nitrérgico e que o α-tocoferol reverte esse efeito comportamental induzido pela cafeína de forma total.

Biogênese mitocondrial e marcadores de estresse oxidativo em ratos treinados e suplementados com L-Arginina

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

Comparação da perfusão vascular folicular, luteal, uterina e perfil hormonal plasmático entre éguas jovens e idosas suplementadas ou não com L-Arginina

Pós-graduação em Biotecnologia Animal - FMVZ

Análises morfológica e histoquímicas do ventrículo e do cérebro de operárias recém emergidas de Apis Mellifera L. (Hymenoptera, Apidae) tratadas com Thiamethoxam

Apis mellifera honeybees are social insects of economic importance, by providing honeybee products, and by the pollination of natural areas of vegetation or agricultural areas. The constant use of pesticides, including the thiamethoxam, which is an insecticide belonging to the class of neonicotinoids with neurotoxic action, is subjecting pollinators to situations of severe stress, which has been evidenced by the decrease in the density of honeybees in many parts of the world. By these considerations, the present study aimed to assess the acute toxicity of thiamethoxam for newly emerged workers of A. mellifera, and to investigate the effect of sublethal doses of this insecticide on the survival time and its cytotoxicity to the brain and midgut to the honeybees. It was established the value of the lethal concentration 50 (LC50) equal to 4.28 ng thiamethoxam/μL of food and from it, the sublethal concentrations of 0.428 ng thiamethoxam/μL (CL50/10) and 0.0428 ng thiamethoxam/μL (CL50/100), which were used in bioassays of intoxication of the honeybees. After the bioassays the bees were dissected and the brain and midguts were collected to analyze possible morphological (staining with Hematoxylin-Eosin) and histochemical alterations (Xylidine Ponceau technique, and Feulgen and PAS reactions) caused by exposure to thiamethoxam, and to calculate the lethal time (LT50) for the workers. The duration of the bioassays was 8 days after beginning of feeding. The results obtained showed that the thiamethoxam is toxic to newly-emerged workers of A. mellifera, causing changes in survival time of individuals. This study also shown that the thiamethoxam causes morphological and histochemical alterations on the midgut and brain of workers. These alterations may be reflected in physiological and behavioral changes that can modify the operation of the colony

Níveis plasmáticos de nitrito/nitrato, GMPc e cortisol em mulheres normotensas e hipertensas após a menopausa: efeito crônico do exercício físico e exposição aguda à L-arginina

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

Aorta from angiotensin II hypertensive mice exhibit preserved nitroxyl anion mediated relaxation responses

Hypertension is a disorder affecting millions worldwide, and is a leading cause of death and debilitation in the United States. It is widely accepted that during hypertension and other cardiovascular diseases the vasculature exhibits endothelial dysfunction; a deficit in the relaxatory ability of the vessel, attributed to a lack of nitric oxide (NO) bioavailability. Recently, the one electron redox variant of NO, nitroxyl anion (NO-) has emerged as an endothelium-derived relaxing factor (EDRF) and a candidate for endothelium-derived hyperpolarizing factor (EDRF). NO- is thought to exist protonated (HNO) in vivo, which would make this species more resistant to scavenging. However, no studies have investigated the role of this redox species during hypertension, and whether the vasculature loses the ability to relax to HNO. Thus, we hypothesize that aorta from angiotensin II (AngII)-hypertensive mice will exhibit a preserved relaxation response to Angeli's Salt, an HNO donor. Male C57B16 mice, aged 12-14 weeks were implanted with mini-osmotic pumps containing AngII (90 ng/min, 14 days plus high salt chow) or sham surgery. Aorta were excised, cleaned and used to perform functional studies in a myograph. We found that aorta from AngII-hypertensive mice exhibited a significant endothelial dysfunction as demonstrated by a decrease in acetylcholine (ACh)-mediated relaxation. However, vessels from hypertensive mice exhibited a preserved response to Angeli's Salt (AS), the HNO donor. To confirm that relaxation responses to HNO were maintained, concentration response curves (CRCs) to ACh were performed in the presence of scavengers to both NO and HNO (carboxy-PTIO and L-cys, resp.). We found that ACh-mediated relaxation responses were significantly decreased in aorta from sham and almost completely abolished in aorta from AngII-treated mice. Vessels incubated with L-cys exhibited a modest decrease in ACh-mediated relaxations responses. These data demonstrate that aorta from AngII-treated hypertensive mice exhibit a preserved relaxation response to AS, an HNO donor, regardless of a significant endothelial dysfunction. (C) 2011 Elsevier Ltd. All rights reserved,