Reduced collagen accumulation and augmented MMP-2 activity in left ventricle of old rats submitted to high-intensity resistance training.

Progressive fibrosis is a hallmark of the aging heart. Age-related fibrosis is modulated by endurance exercise training; however, little is known concerning the influence of resistance training (RT). Therefore we investigated the chronic effects of high-intensity RT on age-associated alterations of left ventricle (LV) structure, collagen content, matrix metalloproteinase-2 (MMP-2), and extracellular matrix-related gene expression, including transforming growth factor-ß (TGF-ß). Young adult (3 mo) and aged (21 mo) male Wistar rats were submitted to a RT protocol (ladder climbing with 65, 85, 95, and 100% load), three times a week for 12 wk. Forty-eight hours posttraining, arterial systolic and diastolic pressure, LV end-diastolic pressure (LVEDP) and dP/dt were recorded. LV morphology, collagen deposition, and gene expression of type I (COL-I) and type III (COL-III) collagen, MMP-2, tissue inhibitor of metalloproteinases-1 (TIMP-1), and TGF-ß1 were analyzed by quantitative reverse transcriptase-PCR. MMP-2 content was assessed by zymography. Increased collagen deposition was observed in LV from aged rats. These parameters were modulated by RT and were associated with increased MMP-2 activity and decreased COL-I, TGF-ß1, and TIMP-1 mRNA content. Despite the effect of RT on collagen accumulation, there was no improvement on LVEDP and maximal negative LV dP/dt of aged rats. Cardiomyocyte diameter was preserved in all experimental conditions. In conclusion, RT attenuated age-associated collagen accumulation, concomitant to the increase in MMP-2 activity and decreased expression of COL-I, TGF-ß1, and TIMP-1 in LV, illustrating a cardioprotective effect of RT on ventricular structure and function.NEW & NOTEWORTHY We demonstrated the beneficial resistance-training effect against age-related left ventricle collagen accumulation in the left ventricle, which was associated with decreased type I collagen (COL-I), transforming growth factor-ß1 (TGF-ß1), and tissue inhibitor of metalloproteinases-1 (TIMP-1) gene expression and matrix metalloproteinase-2 (MMP-2) activity. Our findings suggest for the first time the potential effects of resistance training in modulating collagen accumulation and possibly fibrosis in the aging heart.

Nitric Oxide Modulates HCN Channels in Magnocellular Neurons of the Supraoptic Nucleus of Rats by an S-Nitrosylation-Dependent Mechanism.

The control of the excitability in magnocellular neurosecretory cells (MNCs) of the supraoptic nucleus has been attributed mainly to synaptic inputs from circunventricular organs. However, nitric oxide (NO), a gaseous messenger produced in this nucleus during isotonic and short-term hypertonic conditions, is an example of a modulator that can act directly on MNCs to modulate their firing rate. NO inhibits the electrical excitability of MNCs, leading to a decrease in the release of vasopressin and oxytocin. Although the effects of NO on MNCs are well established, the mechanism by which this gas produces its effect is, so far, unknown. Because NO acts independently of synaptic inputs, we hypothesized that ion channels present in MNCs are the targets of NO. To investigate this hypothesis, we used the patch-clamp technique in vitro and in situ to measure currents carried by hyperpolarization-activated and nucleotide-gated cation (HCN) channels and establish their role in determining the electrical excitability of MNCs in rats. Our results show that blockade of HCN channels by ZD7288 decreases MNC firing rate with significant consequences on the release of OT and VP, measured by radioimmunoassay. NO induced a significant reduction in HCN currents by binding to cysteine residues and forming S-nitrosothiol complexes. These findings shed new light on the mechanisms that control the electrical excitability of MNCs via the nitrergic system and strengthen the importance of HCN channels in the control of hydroelectrolyte homeostasis. SIGNIFICANCE STATEMENT: Cells in our organism live in a liquid environment whose composition and osmolality are maintained within tight limits. Magnocellular neurons (MNCs) of the supra optic nucleus can sense osmolality and control the synthesis and secretion of vasopressin (VP) and oxytocin (OT) by the neurohypophysis. OT and VP act on the kidneys controlling the excretion of water and sodium to maintain homeostasis. Here we combined electrophysiology, molecular biology, and radioimmunoassay to show that the electrical activity of MNCs can be controlled by nitric oxide (NO), a gaseous messenger. NO reacts with cysteine residues (S-nitrosylation) on hyperpolarization-activated and nucleotide-gated cation channels decreasing the firing rate of MNCs and the consequent secretion of VP and OT.

Transcription factor CREB3L1 mediates cAMP and glucocorticoid regulation of arginine vasopressin gene transcription in the rat hypothalamus.

BACKGROUND: Arginine vasopressin (AVP), a neuropeptide hormone that functions in the regulation of water homeostasis by controlling water re-absorption at kidneys, is synthesised in supraoptic nucleus and paraventricular nucleus of the hypothalamus. An increase in plasma osmolality stimulates secretion of AVP to blood circulation and induces AVP synthesis in these nuclei. Although studies on mechanism of AVP transcriptional regulation in hypothalamus proposed that cAMP and glucocorticoids positively and negatively regulate Avp expression, respectively, the molecular mechanisms have remained elusive. Recently, we identified CREB3L1 (cAMP-responsive element binding protein 3 like 1) as a putative transcription factor of Avp transcription in the rat hypothalamus. However the mechanism of how CREB3L1 is regulated in response of hyperosmotic stress in the neurons of hypothalamus has never been reported. This study aims to investigate effect of previously reported regulators (cAMP and glucocorticoid) of Avp transcription on transcription factor CREB3L1 in order to establish a molecular explanation for cAMP and glucocorticoids effect on AVP expression. RESULTS: The effect of cAMP and glucocorticoid treatment on Creb3l1 was investigated in both AtT20 cells and hypothalamic organotypic cultures. The expression of Creb3l1 was increased in both mRNA and protein level by treatment with forskolin, which raises intracellular cAMP levels. Activation of cAMP by forskolin also increased Avp promoter activity in AtT20 cells and this effect was blunted by shRNA mediated silencing of Creb3l1. The forskolin induced increase in Creb3l1 expression was diminished by combined treatment with dexamethasone, and, in vivo, intraperitoneal dexamethasone injection blunted the increase in Creb3l1 and Avp expression induced by hyperosmotic stress. CONCLUSION: Here we shows that cAMP and glucocorticoid positively and negatively regulate Creb3l1 expression in the rat hypothalamus, respectively, and regulation of cAMP on AVP expression is mediated through CREB3L1. This data provides the connection between CREB3L1, a newly identified transcription factor of AVP expression, with the previously proposed mechanism of Avp transcription which extends our understanding in transcription regulation of Avp in the hypothalamus.

Neurohypophyseal response to fluid resuscitation with hypertonic saline during septic shock in rats.

Septic shock is a serious condition with a consequent drop in blood pressure and inadequate tissue perfusion. Small-volume resuscitation with hypertonic saline (HS) has been proposed to restore physiological haemodynamics during haemorrhagic and endotoxic shock. In the present study, we sought to determine the effects produced by an HS infusion in rats subjected to caecal ligation and perforation (CLP). Male Wistar rats were randomly grouped and submitted to either CLP or sham surgery. Either HS (7.5% NaCl, 4 ml kg(-1) i.v.) or isotonic saline (IS; 0.9% NaCl, 4 ml kg(-1) i.v.) was administered 6 h after CLP. Recordings of mean arterial pressure and heart rate were made during this protocol. Moreover, measurements of electrolyte, vasopressin and oxytocin secretion were analysed after either the HS or the IS treatment. Six hours after CLP, we observed a characteristic decrease in mean arterial pressure that occurs after CLP. The HS infusion in these rats produced a transient elevation of the plasma sodium concentration and osmolality and increased plasma vasopressin and oxytocin levels. Moreover, the HS infusion could restore the mean arterial pressure after CLP, which was completely blunted by the previous injection of the vasopressin but not the oxytocin antagonist. The present study demonstrated that rats subjected to CLP and an infusion of hypertonic saline respond with secretion of neurohypophyseal hormones and a transient increase in blood pressure mediated by the V(1) receptor.

Oxido nítrico e peptídeo atrial natriurético na predição de complicações da gestação / Nitric oxide and atrial natriuretic peptide in the prediction of pregnancy complications

OBJETIVO: verificar a acurácia das dosagens séricas maternas do peptídeo atrial natriurético (ANP) e óxido nítrico (NO) para predição de complicações da gravidez. MÉTODOS: a casuística compreendeu 49 mulheres primigestas. As gestantes foram incluídas no estudo na 18ªsemana, momento em que foi coletada a amostra sangüínea para a realização das dosagens séricas. O ANP foi dosado pelo método de radioimunoensaio, utilizando kits Euro-dianostica (2000), considerando anormais valores superiores a 237,4 pg/mL (percentil 95). A dosagem do NO foi realizada pelo método de quimiluminescência, sendo considerados como anormais valores superiores a 17,8 µmol/L (percentil 95). Para análise estatística, utilizaram-se o teste t não pareado para a análise das variáveis quantitativas contínuas de distribuição normal; o teste de Mann-Whitney para amostras quantitativas não-paramétricas; o teste exato de Fisher na avaliação dos parâmentros qualitativos; e o teste de Pearson na avaliação das correlações. RESULTADOS: os dados não mostraram diferença significativa na concentração sérica do ANP, considerando o grupo que apresentou complicações gestacionais e/ou perinatais (média de 139,3±77,1 pg/mL) e o grupo controle (média de 119,6±47,0 pg/mlL), e nem na concentração sérica do NO, entre o grupo com complicações gestacionais e/ou perinatais (média de 11,1±4,6 æmol/L) e o grupo controle (média de 10,0±3,4 µmol/L). CONCLUSÕES: os resultados mostram que o ANP e o NO não foram bons indicadores de complicações da gestação.

PURPOSE: to verify the effectiveness of the maternal blood serum assays of the atrial natriuretic peptide (ANP) and nitric oxide (NO) to predict pregnancy complications. METHODS: the sample was made of 49 primigravidae women. They were included in the study at the 18th week of gestation, when blood sample was collected in order to analyze the serum assays. ANP was assayed by radioimmunoassay, using Euro-dianostica kits (2000), considering abnormal values over 237.4 pg/ml (95 percentil). NO level was evaluated by the chemiluminescence method, considering abnormal values over 17.8 mmol/l (percentil 95). For the statistical analysis of continuous quantitative variables with normal distribution, the unpaired t test was used; Mann-WhitneyÆs test was used for non parametrical quantitative samples; FisherÆs exact test, for the qualitative parameter assessment; and PearsonÆs test for the assessment of correlations. RESULTS: there was no significant difference in the blood serum concentration of ANP between the group that presented complications during pregnancy and/or peridelivery (139.3±77.1 pg/ml) and the control group (119.6±47.0 pg/ml), nor in the serum concentration of NO, either, among the ones with complications in the pregnancy and/or in the peridelivery (11.1±4,6 mmol/l) and the control group (10.0±3.4 mmol/l). CONCLUSIONS: the results show that ANP and NO serum levels are not good predictors of pregnancy complications.

Estradiol reduces cumulative mercury and associated disturbances in the hypothalamus-pituitary axis of ovariectomized rats.

The aim of this research was to verify the incidence of endocrine dysfunction associated with mercury intoxication in the hypothalamus-pituitary reproductive system of normally cycling or castrated female rats and the possible protective action of estrogen replacement therapy. We found no differences in the frequency of estrus cycle stages (diestrus I, diestrus II, proestrus, and estrus) in normally cycling female rats during 54 days of daily oral administration of 0.004, 0.02, and 1 mg/kg MeHgCl. Conversely, the higher dose (1 mg/kg) induced a significant decrease in content of luteinizing hormone releasing hormone (LHRH) into the medial hypothalamus when administered daily during 3 days in ovariectomized rats. This effect was associated with increased levels of mercury found in the anterior pituitary gland and medial hypothalamus, rather than the anterior and posterior hypothalamus, striatum or cerebellum. A decrease in plasma levels of luteinizing hormone (LH) was also detected after administration of 7.5 mg/kg MeHgCl. These disturbances in LHRH and LH secretion induced by mercury were abolished or superimposed (respectively) by estrogenic replacement therapy (0.025 mg/kg 17beta estradiol cypionate, intramuscular). These effects were associated with a significant reduction in mercury content of the anterior pituitary gland and medial hypothalamus, suggesting a protective estrogenic effect.

Interaction between supraoptic nucleus and septal area in the control of water, sodium intake and arterial blood pressure induced by injection of angiotensin II.

We investigated the effects of injection into the supraoptic nucleus (SON) of losartanand PD 123319 (nonpeptide AT(1) and AT(2)-angiotensin II [ANG II] receptor antagonists, respectively); d(CH(2))(5)-Tyr(Me)-AVP (AVPA; an arginine-vasopressin [AVP] V(1) receptor antagonist), FK 409 (a nitric oxide [NO] donor), and N(W)-nitro-l-arginine methyl ester (l-NAME; an NO synthase inhibitor) on water intake, sodium chloride 3% (NaCl) intake and arterial blood pressure induced by injection of ANG II into the lateral septal area (LSA). Male Holtzman rats (250-300 g) were implanted with cannulae into SON and LSA unilaterally. The drugs were injected in 0.5 microl over 30-60 s. Controls were injected with a similar volume of 0.15 M NaCl. ANG II was injected at a dose of 10 pmol. ANG II antagonists and AVPA were injected at doses of 80 nmol. FK 409 and l-NAME were injected at doses of 20 and 40 microg, respectively. Water and NaCl intake was measured over a 2-h period. Prior administration of losartan into the SON decreased water and NaCl intake induced by injection of ANG II. While there was a decrease in water intake, ANG II-induced NaCl intake was significantly increased following injection of AVPA. FK 409 injection decreased water intake and sodium intake induced by ANG II. l-NAME alone increased water and sodium intake and induced a pressor effect. l-NAME-potentiated water and sodium intake induced by ANG II. PD 123319 produced no changes in water or sodium intake induced by ANG II. The prior administration of losartan or AVPA decreased mean arterial pressure (MAP) induced by ANG II. PD 123319 decreased the pressor effect of ANG II to a lesser degree than losartan. FK 409 decreased the pressor effect of ANG II while l-NAME potentiated it. These results suggest that both ANG II AT(1) and AVP V(1) receptors and NO within the SON may be involved in water intake, NaCl intake and the pressor response were induced by activation of ANG II receptors within the LSA. These results do not support the involvement of LSA AT(2) receptors in the mediation of water and NaCl intake responses induced by ANG II, but influence the pressor response.

On the purinergic regulation of hormonal secretion from the anterior pituitary gland

Purinergic regulation of hormonal secretion from the anterior pituitary may be characterized by effects with biphasic secretory response. This response may be started by activation of different subtypes of membrane prurinergic receptor (A1 and/or A2). A putative autocrine mechanism has been proposed to explain the action of adenosine on pituitary hormonal secretion. This mechanism may be dependent on adenosine degradation by the enzyme adenosine deaminase into the extracellular space. The regulation of AMPc and calcium levels in cytoplasm may be part of putative intracellular mechanisms involved in purinergic action. Additionally, hypophysiotrophic effects induced by hypothalamic substances may be modulated by adenosine. The mechanisms involved in this modulatory effects, however, remain elusive.

Faculdade de Medicina de Ribeiräo Preto / Faculdade de Medicina de Ribeiräo Preto

Apresenta a história da Faculdade de Medicina de Ribeiräo Preto. Afirma que a fundaçäo da Faculdade em 1952 foi resultado do crescimento demográfico do Estado de Säo Paulo e representou o primeiro passo da Universidade de Säo Paulo em direçäo ao interior do Estado. Em sua organizaçäo, a Faculdade foi pioneira em adotar normas näo comuns aos estabelecimentos universitários no Brasil. (JGC)

Participation of the ß-adrenoceptors of the medial septal area on urine flow rate, sodium and potassium excretion

Investigou-se a participaçäo dos ß-adrenoceptores da área septal (AS) na excreçäo urinária de sódio, potássio e fluxo urinário. As alteraçöes na pressäo arterial e algumas funçöes renais foram também investigadas. A injeçäo de 2.10-8 à 16.10-8 M de isoproterenol, através de cânulas de demora implantadas permanentemente na AS, produziu uma diminuiçäo significante dose-dependente na excreçäo urinária de sódio, potássio e fluxo urinário. Pré-tratamento com 16.10-8 M de butoxamina antagonizou o efeito de 4.10-8 M de isoproterenol mas o pré-tratamento com 16.10-8 M de practolol näo aboliu o efeito do isoproterenol. Os ß2 agonistas, terbutalina e salbutamol (4.10-8 M) quando injetados intraseptalmente produziram também um decréscimo no fluxo urinário e na excreçäo de sódio e potássio. Após injeçäo de isoproterenol ou salbutamol (4.10-8 M) na AS, a pressäo arterial, taxa de filtraçäo glomerular (TFG) e a filtraçäo de sódio foram reduzidas, enquanto que a fraçäo de reabsorçäo de sódio foi aumentada. Os resultados indicam que os ß2 adrenoceptores da AS tem influência na diminuiçäo de sódio, potássio e fluxo urinário e este efeito pode ser devido a queda na TFG e no sódio filtrado e pelo aumento na reabsorçäo tubular de sódio

Effect of brain natriuretic peptide on dehydration- or angiotensin II- induced water intake in rats

The present study was performed to evaluate the effect of 3 rdV injection on water intake of brain natriuretic peptide (BNP), which is structurally different from the atrial natriuretic peptide. VNP was recently isolated from porcine brain and appears to have a different precursor than the family of atrial natriuretic peptides. Central administration of BNP 3rdV decreased water intake. At a dose of 2.0 mmol/rat, BNP partially inhibited dehyfration-induced water intake and completely blocked the stimulatory effect of 478 pmol/rat angiotension II in rats

Effect of adenosine on gonadotropin and prolactin secretion by hemipituitaries in vitro

Incubation of hemipituitaries from male rats (200-220 g) with 10 nM to 1 micronM adenosine induced a dose-dependent decrease of the release of luteinizing hormone (LH) and follicle stimulating hormone (FSH) into the medium, and increased prolactin (PRL) secretion. The effects of 10 nM adenosine were blocked by 100 nM ceffeine, wereas 100 nM ceffeine alone had also inhibited 10 nM luteinizing hormone-releasing hormone (LHRH)-induced LH and FSH release by > 90%. These data indicate a regulatory role for adenosine in pituitary LH, FSH and PRL release, and also a possible modulatory effect of adenosine on the LHRH-LH and FSH system

Role of adrenals in the increase of sodium chloride intake following lesions in the septal area of the rat

Foi investigado o papel das adrenais na ingestäo da soluçäo de cloreto de sódio aumentada, por lesäo bilateral da área septal. Observou-se que quando a lesäo provocou um aumento grande da ingestäo de cloreto de sódio, a adrenalectomia determinou uma reduçäo nesta ingestäo. Por outro lado, quando o aumento foi pequeno, a adrenalectomia provocou um aumento maior. Em ratos adrenalectomizados e com ingestäo mantida por terapêutica hormonal substitutiva, a lesäo da área septal provocou um aumento no consumo de cloreto de sódio. Este resultado descarta a possibilidade de se atribuir a uma variaçäo de secreçäo hormonal da adrenal, por lesäo da área septal, desde que os níveis de corticosteróides foram mantidos constantes pela hormônio terapia substitutiva. Quando se efetuou a lesäo septal e a adrenalectomia concomitantemente, a ingestäo foi aumentada, e näo foi reduzida pela terapêutica hormonal substitutiva, enquanto que nos animais apenas adrenalectomizados esta ingestäo foi diminuída. Estes resultados mostraram que o aumento da ingestäo de cloreto de sódio provocado por lesäo septal, näo depende primariamente das adrenais

Substance P injected into the medial preoptic area inhibits sodium, potassium and water urinary excretion: putative modulation of the cholinergic system

Substance P (SP, 1.5 nmol) injected into the medial preoptic area (MPOA) of conscious, unrestrained, water-loaded male rats induced a significant decrease in urinary sodium, potassium and water excretion. In contrast, a significant natriuretic effect was observed after injection of 0.3 nmol of a specific competitive SP antagonist ([D-Pr**2, D-Trp**7,**9]-substance P). SP partially blocked the carbachol-induced natriuresis in a time-dependent manner. These data indicate a tonic inhibitory action of SP on sodium excretion and suggest a putative modulatory action of SP on the cholinergic system

O efeito de hormônios neurohipofisários sobre a excreçäo de eletrólitos em ratos / The effect of neurohypophyseal hormones on the excretion of electrolytes in rats

Os autores estudaram o efeito da infusäo endovenosa de uma soluçäo aquosa de HAD e ocitocina, em diferentes períodos de tempo. Pela análise dos resultados verificaram que ambos os hormônios alteram significantemente a excreçäo urinária de água e sódio. Contudo, a excreçäo renal de potássio nem sempre acompanhou o pico de excreçäo de sódio. Os dados sugerem uma inibiçäo da reabsorçäo de sódio ao nível do túbulo proximal para distal