Evidences of antihypertensive potential of extract from Solanum capsicoides All. in spontaneously hypertensive rats.

BACKGROUND: Solanum capsicoides All. is morphologically similar to Solanum sisymbriifolium Lam. which is used in folk medicine in South America for antihypertensive and diuretics purposes. This similarity has led to species identification errors, which therefore may result in errors by patients. PURPOSE: To evaluate the antihypertensive and diuretics potential of the methanol extract from Solanum capsicoides All. (MeOH-Sc), in vitro and in vivo, in spontaneously hypertensive rats (SHR). METHODS: Initial experiments were performed in rat mesenteric artery to evaluate the in vitro vascular effect of MeOH-Sc and its fractions, in addition to the mechanisms involved during the observed effect. Mean arterial pressure (MAP) and heart rate (HR) were recorded in non-anesthetised hypertensive and normotensive rats. In another set of experiments, MeOH-Sc was administered for 21 consecutive days. Daily body weight measurements were conducted and MAP, HR and urinary volume were measured every 5 days. The mesenteric artery from treated animals was tested for phenylephrine and sodium nitroprussiate (SNP) sensitivity. RESULTS: Initially, MeOH-Sc and fractions relaxed phenylephrine-induced contractions in mesenteric artery rings. The vasorelaxant effect was not changed in the presence of a blocker of eNOS (L-NAME) in rings with an intact endothelium. In denuded-endothelium rings, the vasorelaxant response was significantly reduced in the presence of a cAMP inhibitor (SQ 22536 10 µM) in SHR but not in Wistar Kyoto rats (WKY). However, in the presence of a cGMP inhibitor (ODQ 10 µM), a curve shift to the right was observed in WKY animals, but not in SHR. Intravenous bolus injections of MeOH-Sc into non-anesthetised SHR and WKY, induced hypotension that was associated with an increase in HR. A significant antihypertensive effect was observed in animals that received MeOH-Sc orally for 21 days, which also prevented the development of cardiac hypertrophy. Urine volume from animals treated with MeOH-Sc significantly increased. Finally, MeOH-Sc induced beneficial changes in vascular responsiveness. CONCLUSION: MeOH-Sc has a potential antihypertensive effect in SHR.

Blockade of central delta-opioid receptors inhibits salt appetite in sodium-depleted rats.

Various studies have investigated the role of central opioid peptides in feeding behavior; however, only a few have addressed the participation of opioids in the control of salt appetite. The present study investigated the effect of intracerebroventricular injections of the δ-opioid antagonist, naltrindole (5, 10 and 20 nmol/rat) and the agonist, deltorphin II (2.5, 5, 10 and 20 nmol/rat) on salt intake. Two protocols for inducing salt intake were used: sodium-depletion and the central injection of angiotensin II. In addition, the effect of a central δ-opioid receptor blockade on locomotor activity, on palatable solution intake (0.1% saccharin) and on blood pressure was also studied. The blockade of central δ-opioid receptors inhibits salt intake in sodium-depleted rats, while the pharmacological stimulation of these receptors increases salt intake in sodium-replete animals. Furthermore, the blockade of central δ-opioid receptors inhibits salt intake induced by central angiotensinergic stimulation. These data suggest that during sodium-depletion activation of the δ-opioid receptors regulates salt appetite to correct the sodium imbalance and it is possible that an interaction between opioidergic and angiotensinergic brain system participates in this control. Under normonatremic conditions, δ-opioid receptors may be necessary to modulate sodium intake, a response that could be mediated by angiotensin II. The decrease in salt intake following central δ-opioid receptors blockade does not appear to be due to a general inhibition of locomotor activity, changes in palatability or in blood pressure.

Central kappa opioid receptors modulate salt appetite in rats.

The role of the central opioid system in the control of water and salt intake is complex, with both stimulatory and inhibitory effects having been observed. The aim of the present study was to investigate the participation of the central κ-opioid receptors in the control of salt appetite. Male Wistar rats were submitted to two different experimental protocols: sodium deficit produced by the diuretic, furosemide, and brain angiotensinergic stimulation in rats under normal sodium balance. Lateral ventricle (LV) injections of Nor-binaltorphimine (Nor-BNI) at different doses (5, 10 and 20 nmol) inhibited hypertonic saline solution (1.5%) intake in sodium-depleted rats. The salt appetite induced by an LV injection of angiotensin II (Ang II) (10 ng) was also blocked by Nor-BNI injections into the LV, while no significant change was observed in water intake. Furthermore, the decrease in salt intake seems not to have been due to a general inhibition of locomotor activity or to any change in palatability, since central administration of Nor-BNI failed to modify the intake of a 0.1% saccharin solution when the animals were submitted to a "dessert test" or to induce any significant locomotor deficit in the open-field test. Also the central administration of Nor-BNI was unable to modify blood pressure in sodium-depleted animals. The present results suggest that activation of endogenous κ-opioid receptors modulates salt appetite induced by sodium depletion and by central angiotensinergic stimulation in rats.

Multiple opioid receptors mediate the hypotensive response induced by central 5-HT(3) receptor stimulation.

The aim of the present work was to investigate the role of brain µ, κ and δ opioid receptors in the central serotonergic mechanisms regulating blood pressure in rats. The data obtained show that: (1) pharmacological activation of central 5-HT(3) receptors yields a significant decrease in blood pressure; (2) the blockade of those receptors by a selective antagonist induces an acute hypertensive response; (3) the pharmacological blockade of central opioid receptors by three different opioid antagonists exhibiting variable degrees of selectivity to µ, κ and δ opioid receptors always suppressed the hypotensive response induced by central 5-HT(3) receptor stimulation; (4) the blockade of opioid receptors by the same opioid antagonists that impaired the hypotensive effect of central 5-HT(3) receptor stimulation failed to modify blood pressure in animals not submitted to pharmacological manipulations of central 5-HT(3) receptor function. It is shown that a 5-HT(3) receptor-dependent mechanism seems to be part of the brain serotonergic system that contributes to cardiovascular regulation since the hypertensive response observed after ondansetron administration indicates that central 5-HT(3) receptors exert a tonic inhibitory drive on blood pressure. Furthermore, the data obtained here clearly indicate that the hypotensive response observed after pharmacological stimulation of central 5-HT(3) receptors depends on the functional integrity of brain µ, κ and δ opioid receptors, suggesting that a functional interaction between serotonergic and opiatergic pathways in the brain is part of the complex, multifactorial system that regulates blood pressure in the central nervous system.

Blockade of central kappa-opioid receptors inhibits the antidipsogenic effect of interleukin-1beta.

The objective of the present study was to investigate the role of brain kappa-opioid receptors (KOR) in the antidipsogenic effect promoted by third ventricle injections of interleukin-1beta (IL-1beta). Wistar male rats were submitted to three different, thirst-inducing, physiological conditions: dehydration induced by water deprivation, hyperosmolarity induced by salt-load and hypovolemia induced by polyethylene glycol subcutaneous injection. Third ventricle injections of IL-1beta significantly inhibited the increase in water intake observed in those situations. The pharmacological blockade of central KOR by the selective KOR antagonist nor-binaltorphimine (BNI) at different doses significantly inhibited the antidipsogenic effect induced by the central administration of IL-1beta in all conditions tested: dehydration, hypovolemia and hyperosmolarity. The central administration of IL-1beta failed to induce any locomotor deficit, as verified in an open field test. Stimulation of the central interleukinergic component did not result in any general suppression of ingestive behavior since no change in saccharin intake was recorded during a dessert test in animals receiving central injections of IL-1beta. Furthermore, the inhibitory effects of IL-1beta on water intake cannot be attributed to sickness-like effects induced by these compounds, since an aversion test excluded this possibility. In summary, the data shown in the present study clearly show that the antidipsogenic effect observed in rats following third ventricle injections of IL-1beta depend on the functional integrity of a brain kappa-opioid-dependent component.

The central amygdala regulates sodium intake in sodium-depleted rats: role of 5-HT3 and 5-HT2C receptors.

In the present paper, we have evaluated the participation of 5-HT(3) and 5-HT(2C) receptors in the central amygdala (CeA) in the regulation of water and salt intake in sodium-depleted rats. m-CPBG-induced pharmacological activation of 5-HT(3) receptors located in the CeA resulted in a significant reduction in salt intake in sodium-depleted rats. This antinatriorexic effect of m-CPBG was reverted by pretreatment with the selective 5-HT(3) receptor antagonist ondansetron. The injection of ondansetron alone into the CeA had no effect on sodium-depleted and normonatremic rats. Conversely, pharmacological stimulation of 5-HT(2C) receptors located in the central amygdala by the selective 5-HT(2C) receptor agonist m-CPP failed to modify salt intake in sodium-depleted rats. Additionally, the administration of a selective 5-HT(2C) receptor blocker, SDZ SER 082, failed to modify salt intake in rats submitted to sodium depletion. These results lead to the conclusion that the pharmacological activation of 5-HT(3) receptors located within the CeA inhibits salt intake in sodium-depleted rats and that 5-HT(2C) receptors located within the CeA appear to be dissociated from the salt intake control mechanisms operating in the central amygdala.

Role of the central opioid system in the inhibition of water and salt intake induced by central administration of IL-1beta in rats.

In the present study we investigated, the effect of third ventricle injections of IL-1beta on water and salt intake in fluid-deprived and sodium-depleted rats. Central administration of IL-1beta significantly reduced water and salt intake in fluid-deprived animals and decreased salt intake in sodium-depleted rats. The antidipsogenic and antinatriorexic effects elicited by the central administration of IL-1beta were suppressed by pretreatment with central injections of the non-selective opioid antagonist naloxone (10 mug) in the two different experimental protocols used here (water deprivation and sodium depletion). In addition, central administration of IL-1beta failed to modify the intake of a 0.1% saccharin solution when the animals were submitted to a "dessert test" or to induce any significant locomotor deficit in the open-field test. The present results suggest that the activation of the central interleukinergic component by IL-1beta impairs the increase in water and salt intake induced by water deprivation and the enhancement in sodium appetite that follows sodium depletion. The data also support the conclusion that the antidipsogenic and antinatriorexic effects resulting from the activation of the central interleukinergic component rely on an opioid-dependent, naloxone-blockable system.

Inhibition of water intake by the central administration of IL-1beta in rats: role of the central opioid system.

In the present study we investigated, the effect of third ventricle injections of IL-1beta on water intake, in rats, induced by three different physiological stimuli: dehydration induced by water deprivation, hypernatremia associated with hyperosmolarity induced by intragastric salt load, and hypovolemia produced by subcutaneous polytethyleneglycol administration. Central administration of IL-1beta at the doses of 4 and 8 ng reduced water intake in all three conditions studied. Third ventricle injections of IL-1beta (8 ng) were unable to diminish water intake in the groups of rats pretreated with central injections of the non-selective opioid antagonist naloxone (10 microg) in the three different conditions studied. Furthermore, the central administration of IL-1beta was neither able to modify the intake of a 0.1% saccharin solution when the animals were submitted to a "dessert test" nor to induce any significant locomotor deficit in the open-field test. These results suggest that the central activation of interleukin-1 receptors by IL-1beta is able to impair the thirst-inducing mechanisms triggered by the physiological stimuli represented by dehydration, hyperosmolarity and hypovolemia. These results lead us to conclude that the antidipsogenic effects observed following central administration of IL-1beta require the functional integrity of the brain opiatergic system.

Hyperglycemia induced by pharmacological activation of central serotonergic pathways depends on the functional integrity of brain CRH system and 5-HT3 receptors.

In the present study, we investigated the effect of central serotonergic pathway activation achieved through third ventricle injections of quipazine, a serotonergic agonist, on plasma glucose levels of fasted and fed adult Wistar male rats, whose third ventricles were canulated 7 days before the experiments. Central quipazine administration induced a significant increase in plasma glucose levels in fasted animals, but was unable to modify plasma glucose concentrations in fed rats. Pretreatment with alpha-helical CRH, a CRH antagonist, significantly attenuated quipazine-induced hyperglycemia. Pretreatment with two different 5-HT3 receptor antagonists, LY-278,584 and ondansetron, was also able to produce a significant reduction in the hyperglycemic response evoked by central administration of quipazine. None of the antagonists used was capable of modifying plasma glucose concentrations when injected alone into the third ventricle. Quipazine-treated, hyperglycemic animals did not show any increase in plasma insulin levels. We conclude that acute pharmacological serotonergic stimulation by quipazine produces hyperglycemia by mechanisms that require the functional integrity of both CRH and 5-HT3 receptors, and that impairment in insulin secretion and/or activity may explain hyperglycemia induced by third ventricle injections of quipazine.

Acute fluoxetine administration differentially affects brain C-Fos expression in fasted and refed rats.

In the present study we investigated the effect of acute fluoxetine administration on the expression of c-Fos in the rat brain under two different metabolic conditions: fed and fasting states. Wistar male rats, weighing 220+/-30g, received i.p. injections of saline solution or fluoxetine (10mg/kg), and were killed 2 h later. The brains were removed after transcardiac perfusion with phosphate-buffered saline followed by paraformaldehyde, and were then processed for immunohistochemistry. Fos-like immunoreactivity was quantified by a computerized system. Fasted animals faced an 18-h suppression of food intake, while fed groups were submitted to an initial 14-h period of fast followed by a 4-h period in which food was freely available. Both in fasting and fed states, fluoxetine-treated animals presented a significant increase in c-Fos expression in hypothalamic areas, limbic structures, circumventricular areas, and in mesencephalic and rhomboencephalic regions, as compared with saline-treated controls. The quantitative comparison of data obtained from fasted and fed animals showed that fasted rats treated with fluoxetine presented a higher c-Fos expression in the ventromedial hypothalamus and the paraventricular nuclei compared with the fed group, while in fluoxetine-treated fed rats c-Fos expression was higher in the arcuate nuclei, medial amygdala, locus coeruleus and dorsal raphe nuclei, as compared with fasted, fluoxetine-treated animals. These data indicate that the metabolic condition of the animals significantly modifies fluoxetine-induced brain c-Fos expression, suggesting that visceral and behavioral fluoxetine effects may be influenced by the metabolic state of the individual.

Hyperglycemia induced by acute central fluoxetine administration: role of the central CRH system and 5-HT3 receptors.

Brain serotonin and CRH systems participate in the control of blood glucose levels. We have previously demonstrated that the pharmacological stimulation of central 5-HT3 receptors, the target for several therapeutic agents used as antiemetics in the course of chemotherapy, induces hyperglycemia. The aim of the present study was to investigate the participation of the brain CRH component and 5-HT3 receptors in basal blood glucose levels as well as in the hyperglycemia induced by third ventricle injections of fluoxetine, a serotonin reuptake inhibitor with a broad range of clinical use. In this study, we used fasted adult Wistar male rats (220 +/- 20 g) whose third ventricles were cannulated 7 days prior to the experiments. Acute third ventricle injections of fluoxetine caused a significant increase in plasma glucose levels throughout the experiment. Pretreatment with alpha-helical CRH, a selective CRH antagonist, significantly blunted fluoxetine-induced hyperglycemia. Also, pretreatment with two distinct selective 5-HT3 receptor antagonists (LY-278,584 and ondansetron) significantly impaired the rise in plasma glucose levels observed in fluoxetine-treated animals pretreated with isotonic saline solution. None of these antagonists was able to modify blood glucose levels when injected alone into the third ventricle. Animals receiving third ventricle injections of fluoxetine, in spite of being hyperglycemic, presented plasma insulin levels similar to those displayed by normoglycemic, saline-treated controls. It is suggested that the acute increase in brain serotonergic activity caused by third ventricle injections of fluoxetine induces a hyperglycemic response that requires the functional integrity of the brain CRH system and 5-HT3 receptors. Also, it is proposed that the absence of a compensatory increase in plasma insulin levels may contribute to the generation of a hyperglycemic response after central fluoxetine administration.

Central administration of mCPP, a serotonin 5-HT(2B/2C) agonist, decreases water intake in rats.

In the present study, we investigated in rats the effect of third ventricle injections of 1-(3-chlorophenyl)piperazine (mCPP), a 5-HT(2) receptor agonist, on water intake induced by three different physiological stimuli: fluid deprivation, acute salt load and hypovolemia. Injections of mCPP in the doses of 80 and 160 nmol/rat were able to decrease water intake in all three conditions studied. Third ventricle injections of mCPP (160 nmol/rat) were no longer able to diminish water intake in the groups of rats pretreated with central injections of an equimolar amount of (+)-cis-4,5,7a,8,9,10,11,11a-octahydro-7H-10-methylindolo[1,7-bc][2,6]-naphthyridine (SDZ SER 082), a selective 5-HT(2B/2C) antagonist. The central administration of mCPP (160 nmol/rat) was not able to modify the intake of a 0.1% saccharin solution. It is suggested that the central activation of a 5-HT(2B/2C) component is able to impair the drive for water intake induced by the physiological stimuli represented by fluid deprivation, acute salt load and hypovolemia. This effect seems not to be consequent on a general nonspecific central nervous system depression or on a locomotor deficit, because saccharin intake is not affected by third ventricle injections of mCPP.

Central 5-HT3 receptor stimulation by m-CPBG increases blood glucose in rats.

The aim of the present study was to investigate the role of central 5-HT3 receptors on the control of blood glucose in stressed and non-stressed rats in both fasted and fed states. Adult Wistar male rats had each their third ventricle cannulated 7 days before the experiments. Injections of m-CPBG, a selective 5-HT3 receptor agonist, induced a significant increase in blood glucose in non-stressed rats in both fasted and in fed states. The same procedure was unable to modify stress-induced hyperglycemia. The hyperglycemic effect of m-CPBG central administration was blocked by pretreatment with ondansetron, a specific 5-HT3 receptor antagonist, indicating that the effects here obtained with m-CPBG were a result of its interaction with 5-HT3 receptors. Third ventricle injections of ondansetron alone were not able to modify blood glucose in non-stressed animals and did not change the hyperglycemic responses observed after immobilization stress. We conclude that pharmacological activation of the central 5-HT3 receptor induces a hyperglycemic effect in non-stressed animals.

Effect of the intracerebroventricular administration of GR 113808, a selective 5-HT4 antagonist, on water intake during hyperosmolarity and hypovolemia

We demonstrate here that acute third ventricle injections of GR 113808, a highly selective 5-HT4 antagonist, decrease water intake induced by a previous salt load while potentiating drinking elicited by hypovolemia induced by previous subcutaneous administration of polyethylene glycol in male Wistar rats (200 + or - 20 g). At the dose of 160 nmol/rat, third ventricle injections of GR 113808 induced a significant reduction of water intake in salt-loaded animals after 120 min as compared to salt-loaded animals receiving third ventricle injections of saline (salt load + GR = 3.44 + or - 0.41 ml, N = 12; salt load + saline = 5.74 + or - 0.40 ml, N = 9). At the dose of 80 nmol/rat, GR 113808 significantly enhanced water intake in hypovolemic animals after 120 min as compared to hypovolemic animals receiving third ventricle injections of saline (hypovol + GR = 4.01 + or - 0.27 ml, N = 8; hypovol + saline = 2.41 + or - 0.23 ml, N = 12). We suggest that central 5-HT4 receptors may exert a positive drive on water intake due to hyperosmolarity and a negative input on drinking provoked by hypovolemia.

Effect of the intracerebroventricular administration of GR 113808, a selective 5-HT4 antagonist, on water intake during hyperosmolarity and hypovolemia.

We demonstrate here that acute third ventricle injections of GR 113808, a highly selective 5-HT4 antagonist, decrease water intake induced by a previous salt load while potentiating drinking elicited by hypovolemia induced by previous subcutaneous administration of polyethylene glycol in male Wistar rats (200 +/- 20 g). At the dose of 160 nmol/rat, third ventricle injections of GR 113808 induced a significant reduction of water intake in salt-loaded animals after 120 min as compared to salt-loaded animals receiving third ventricle injections of saline (salt load + GR = 3.44 +/- 0.41 ml, N = 12; salt load + saline = 5.74 +/- 0.40 ml, N = 9). At the dose of 80 nmol/rat, GR 113808 significantly enhanced water intake in hypovolemic animals after 120 min as compared to hypovolemic animals receiving third ventricle injections of saline (hypovol + GR = 4.01 +/- 0.27 ml, N = 8; hypovol + saline = 2.41 +/- 0.23 ml, N = 12). We suggest that central 5-HT4 receptors may exert a positive drive on water intake due to hyperosmolarity and a negative input on drinking provoked by hypovolemia.

Central 5-HT(4) receptors and drinking behavior.

The aim of the present study was to investigate the effect of acute third ventricle injections of two different 5-HT(4) receptor antagonists, GR 113808 and SB 204070, on water intake in different situations. Injections of 80 nmol/rat of both GR 113808 and SB 204070 were unable to modify water intake in normohydrated rats. Pretreatment with GR 113808 (40 and 80 nmol/rat) and SB 204070 (80 and 160 nmol/rat) blunted water intake after third ventricle injections of angiotensin II (9.6 pmol/rat) compared to saline-pretreated controls. Pretreatment with 80 nmol/rat of both antagonists potentiated drinking induced by third ventricle injections of carbachol (11.0 nmol/rat) compared to saline-pretreated control. In all doses employed, none of the compounds was able to modify water intake in dehydrated rats. A separate control test using one-bottle taste aversion paradigm indicated that the reduction in water intake observed in some of the present experiments could not be attributed to a drug-induced malaise. It is suggested that central 5-HT(4) receptors exert a dualistic role on the control of water intake potentiating angiotensin II-induced drinking and inhibiting thirst induced by central cholinergic activation

Zinc and water intake in rats: investigation of adrenergic and opiatergic central mechanisms.

We have demonstrated that central administration of zinc in minute amounts induces a significant antidipsogenic action in dehydrated rats as well as in rats under central cholinergic and angiotensinergic stimulation. Here we show that acute third ventricle injections of zinc also block water intake induced by central ss-adrenergic stimulation in Wistar rats (190-250 g). Central inhibition of opioid pathways by naloxone reverses the zinc-induced antidipsogenic effect in dehydrated rats. After 120 min, rats receiving third ventricle injections of isoproterenol (160 nmol/rat) exhibited a significant increase in water intake (5.78 +/- 0.54 ml/100 g body weight) compared to saline-treated controls (0.15 +/- 0.07 ml/100 g body weight). Pretreatment with zinc (3.0, 30.0 and 300.0 pmol/rat, 45 min before isoproterenol injection) blocked water intake in a dose-dependent way. At the highest dose employed a complete blockade was demonstrable (0.54 +/- 0.2 ml/100 g body weight). After 120 min, control (NaAc-treated) dehydrated rats, as expected, exhibited a high water intake (7.36 +/- 0.39 ml/100 g body weight). Central administration of zinc blocked this response (2.5 +/- 0.77 ml/100 g body weight). Naloxone pretreatment (82.5 nmol/rat, 30 min before zinc administration) reverted the water intake to the high levels observed in zinc-free dehydrated animals (7.04 +/- 0.56 ml/100 g body weight). These data indicate that zinc is able to block water intake induced by central ss-adrenergic stimulation and that zinc-induced blockade of water intake in dehydrated rats may be, at least in part, due to stimulation of central opioid peptides.

Central lead administration inhibits water intake and sodium appetite in rats.

We have demonstrated that acute third ventricle injections of lead acetate (PbAc) exert a powerful antidipsogenic effect and induce a significant increase in renal sodium excretion. In the present study we confirm the antidipsogenic effect of lead and demonstrate that central administration of this metal, in minute amounts, significantly reduces salt intake both during dehydration and after central angiotensinergic stimulation. Adult male Wistar rats had the third ventricle cannulated seven days before the experiments. During this period they had free access to distilled water and hypertonic saline solution (1.5%). After a 24-h period of fluid deprivation, experimental animals received third ventricle injections of PbAc (0.3, N = 8 and 3.0 nmol/rat, N = 14) while controls received sodium acetate (NaAc; 3.0 nmol/rat, N = 10). Rats treated with PbAc at the highest dose showed a significant reduction (P<0.05) both in water and hypertonic saline intake when compared to controls. When the effect of lead administration on angiotensin II-induced water and salt intake was studied, normohydrated animals received third ventricle injections of angiotensin II (9.6 nmol/rat) after pretreatment with 3.0 nmol/rat of PbAc (experimental group, N = 10) or NaAc (controls, N = 8). The group pretreated with PbAc presented a significant reduction (P<0.05) in both water and salt intake compared to controls. Thus, this study confirms the antidipsogenic effect of central lead injections and demonstrates that the presence of lead in the brain exerts a significant inhibition of sodium appetite.