RESUMO
OBJECTIVE: To investigate the effects of regular chilli ingestion on some indicators of metabolic and vascular function. DESIGN: A randomized cross-over dietary intervention study. SETTING: Launceston, Australia. SUBJECTS: Healthy free-living individuals. INTERVENTION: Thirty-six participants (22 women and 14 men), aged 46+/-12 (mean+/-s.d.) years; BMI 26.4+/-4.8 kg/m(2), consumed 30 g/day of a chilli blend (55% cayenne chilli) with their normal diet (chilli diet), and a bland diet (chilli-free) for 4 weeks each. Metabolic and vascular parameters, including plasma glucose, serum lipids and lipoproteins, insulin, basal metabolic rate, blood pressure, heart rate, augmentation index (AIx; an indicator of arterial stiffness), and subendocardial-viability ratio (SEVR; a measure of myocardial perfusion), were measured at the end of each diet. In a sub-study, during week 3 of each dietary period, the vascular responses of 15 subjects to glyceryl-trinitrate (GTN) and salbutamol were also studied. RESULTS: For the whole group, there were no significant differences between any of the measured parameters when compared at the end of the two dietary periods. When analysed separately, men had a lower resting heart rate (P=0.02) and higher SEVR (P=0.05) at the end of the chilli diet than the bland diet. In the sub-study, baseline AIx on the chilli diet was lower (P<0.001) than on the bland diet, but there was no difference in the effects of GTN and salbutamol between the two diets. CONCLUSION: Four weeks of regular chilli consumption has no obvious beneficial or harmful effects on metabolic parameters but may reduce resting heart rate and increase effective myocardial perfusion pressure time in men.
Assuntos
Artérias/efeitos dos fármacos , Artérias/fisiologia , Capsicum , Dieta , Fluxo Pulsátil/fisiologia , Metabolismo Basal/efeitos dos fármacos , Velocidade do Fluxo Sanguíneo/efeitos dos fármacos , Velocidade do Fluxo Sanguíneo/fisiologia , Glicemia/efeitos dos fármacos , Glicemia/metabolismo , Pressão Sanguínea/efeitos dos fármacos , Capsicum/química , Complacência (Medida de Distensibilidade)/efeitos dos fármacos , Estudos Cross-Over , Suplementos Nutricionais , Feminino , Frequência Cardíaca/efeitos dos fármacos , Humanos , Insulina/sangue , Metabolismo dos Lipídeos/efeitos dos fármacos , Lipídeos/sangue , Masculino , Pessoa de Meia-Idade , Oxirredução , Fatores SexuaisRESUMO
Platypus venom contains an isomerase that reversibly interconverts the second amino-acid residue in some peptides between the L-form and the D-form. The enzyme acts on the natriuretic peptides OvCNPa and OvCNPb, and on the defensin-like peptides DLP-2 and DLP-4, but it does not act on DLP-1. While the isomerization of DLP-2 to DLP-4 is inhibited by the amino-peptidase inhibitor amastatin, it is not affected by the leucine amino-peptidase inhibitor bestatin. The enzyme, that is only present in minute quantities in an extract of the venom gland, is thermally stable up to 55 degrees C, and it was found by anion-exchange chromatography to be acidic. Isolation of the isomerase was carried out by combined ion-exchange chromatography and reverse-phase high performance liquid chromatography (HPLC).
Assuntos
Isomerases de Aminoácido/química , Peptídeo Natriurético Tipo C/química , Ornitorrinco , Peçonhas/enzimologia , Isomerases de Aminoácido/antagonistas & inibidores , Isomerases de Aminoácido/isolamento & purificação , Animais , Isomerismo , Peptídeos/química , Ornitorrinco/metabolismo , Inibidores de Proteases/químicaRESUMO
BACKGROUND: Cyclosporin A (CsA)-treated renal transplant recipients (RTR) exhibit relative hyperhomocystinemia and vascular dysfunction. Folate supplementation lowers homocysteine and has been shown to improve vascular function in healthy subjects and patients with coronary artery disease. The aim of this study was to assess the effects of 3 months of folate supplementation (5 mg/day) on vascular function and structure in RTR. METHODS: A double-blind, placebo-controlled crossover study was conducted in 10 CsA-treated RTR. Vascular structure was measured as carotid artery intima media thickness (IMT) and function was assessed as changes in brachial artery diameter during reactive hyperemia (RH) and in response to glyceryl trinitrate (GTN). Function data were analyzed as absolute and percent change from baseline and area under the diameter/time curve. Blood samples were collected before and after supplementation and analyzed for total plasma homocysteine, folate, vitamin B12 and asymmetric dimethyl arginine (ADMA) in addition to regular measures of hemoglobin, hematocrit, mean corpuscular volume (MCV) and serum creatinine. RESULTS: Folate supplementation significantly increased plasma folate by 687% (p < 0.005) and decreased homocysteine by 37% (p < 0.05) with no changes (p > 0.05) in vitamin B12 or ADMA. There were no significant (p > 0.05) changes in vascular structure or function during the placebo or the folate supplementation phases; IMT; placebo pre mean +/- SD, 0.52 +/- 0.12, post 0.50 +/- 0.11; folate pre 0.55 +/- 0.17, post 0.49 +/- 0.20 mm, 5% change in brachial artery diameter (RH, placebo pre 10 +/- 8, post 6 +/- 5; folate pre 9 +/- 7, post 7 +/- 5; GTN, placebo pre 18 +/- 10, post 17 +/- 9, folate pre 16 +/- 9, post-supplementation 18 +/- 8). CONCLUSION: Three months of folate supplementation decreases plasma homocysteine but has no effect on endothelial function or carotid artery IMT in RTR.
Assuntos
Artéria Braquial/efeitos dos fármacos , Artérias Carótidas/efeitos dos fármacos , Ciclosporina/uso terapêutico , Ácido Fólico/uso terapêutico , Imunossupressores/uso terapêutico , Transplante de Rim , Túnica Íntima/efeitos dos fármacos , Túnica Média/efeitos dos fármacos , Artéria Braquial/patologia , Artérias Carótidas/patologia , Estudos Cross-Over , Ciclosporina/sangue , Método Duplo-Cego , Feminino , Ácido Fólico/sangue , Homocisteína/sangue , Homocisteína/efeitos dos fármacos , Humanos , Imunossupressores/sangue , Masculino , Pessoa de Meia-Idade , Resultado do Tratamento , Túnica Íntima/patologia , Túnica Média/patologiaRESUMO
OBJECTIVE: Previous studies investigating associations between serum lipids and renal disease have generally not taken into account dietary intake or physical activity both known to influence circulating lipids. Furthermore, inclusion of patients on HMG-CoA reductase inhibitors may also have influenced findings due to the pleiotropic effect of this medication. Therefore, the aim of this study is to determine the relationships between serum lipids and renal function in a group of patients not taking lipid-lowering medication and taking into account dietary intake and physical activity. METHODS: Data from 100 patients enrolled in the Lipid Lowering and Onset of Renal Disease (LORD) trial were used in this study. Patients were included with serum creatinine > 120 micromol/l, and excluded if they were taking lipid-lowering medication. Unadjusted and adjusted relationships were determined between fasting serum lipid concentrations (total cholesterol, low-density lipoprotein (LDL) cholesterol, high-density lipoprotein (HDL) cholesterol, triglycerides and total cholesterol/HDL ratio) and measures of renal function (estimated glomerular filtration rate (eGFR), creatinine clearance and serum creatinine) and urinary protein excretion. RESULTS: Significant (p < 0.05) negative unadjusted relationships were found between lipids (total cholesterol, LDL and HDL cholesterol) and serum creatinine. In support of these findings, logarithmically-transformed lipids (total cholesterol, LDL and HDL cholesterol) were significantly associated with eGFR and creatinine clearance although the effects were of a smaller magnitude. Adjustment for dietary saturated fat intake and physical activity did not substantially change these effects. CONCLUSION: These data do not support the premise that lipids are associated with renal dysfunction in patients with normocholesterolemia.
Assuntos
Nefropatias/fisiopatologia , Lipídeos/sangue , Pressão Sanguínea , Creatinina/sangue , Dieta , Método Duplo-Cego , Feminino , Taxa de Filtração Glomerular , Humanos , Nefropatias/sangue , Nefropatias/urina , Masculino , Pessoa de Meia-Idade , Atividade Motora , ProteinúriaRESUMO
In order to determine if elevated temperature during vitellogenesis had a detrimental effect on hepatic estrogen receptors of Atlantic salmon (Salmo salar), 3H-estradiol saturation binding analysis, using one- and two-site binding models, was carried out on extracts of hepatic cytosols from fish held at 14, 18 or 22 degrees C over the austral period of peak vitellogenesis (February to April). With one-site binding analysis, no temperature related difference in either receptor affinity (Kd) or number (Bmax) was found at each sampling point, but there was an apparent decrease in both affinity and number at each temperature over the period of the study. However, some analyses, notably at 22 degrees C during February, were best described using a two-site binding model. At this temperature and time, there was a clear separation of binding affinity into high and low components (Kd = 0.67+/-S.E. 0.05 and 20+/-S.E. 5.6 nM, respectively) (n = 4), which suggests that February was a critical time of temperature related hepatic sensitivity to estrogen. These results support those of other studies where we found that February was also a sensitive time with respect to temperature impairment of in vitro follicular estrogen synthesis, and the greatest period of in vivo temperature sensitivity.
Assuntos
Envelhecimento/metabolismo , Fígado/metabolismo , Receptores de Estrogênio/metabolismo , Salmo salar/fisiologia , Temperatura , Vitelogênese/fisiologia , Animais , Sítios de Ligação , Ligação Competitiva , Núcleo Celular/metabolismo , Citosol/metabolismo , Feminino , Modelos Biológicos , Salmo salar/crescimento & desenvolvimento , Salmo salar/metabolismoRESUMO
The binding characteristics of sex steroid binding protein (SBP) were investigated in vitellogenic and nonreproductive female rainbow trout (Oncorhynchus mykiss), black bream (Acanthopagrus butcheri), and greenback flounder (Rhombosolea tapirina). The binding capacity of rainbow trout and black bream SBP was significantly greater in vitellogenic than in nonreproductive-stage fish. A decrease in binding affinity was observed in male trout injected with estradiol (E(2)) compared to control fish. This difference was not observed after partial purification of the SBP by gel filtration and may have resulted from competitive inhibition of E(2) binding by vitellogenin. No differences in flounder SBP binding characteristics were observed.
Assuntos
Linguado/metabolismo , Oncorhynchus mykiss/metabolismo , Perciformes/metabolismo , Globulina de Ligação a Hormônio Sexual/metabolismo , Vitelogênese , Animais , Ligação Competitiva , Estradiol/metabolismo , Estradiol/farmacologia , Feminino , Hidrocortisona/metabolismo , Hidroxiprogesteronas/metabolismo , Testosterona/metabolismo , Vitelogeninas/metabolismoRESUMO
1. In the present study, the effects of the novel vanilloid agonist, 12-phenylacetate 13-acetate 20-homovanillate (PPAHV), on oxygen consumption (VO(2)) and vascular resistance (perfusion pressure, PP) were investigated in the constant flow, perfused rat hindlimb. The acute desensitizing properties of this novel synthetic agent were also examined. 2. Maximum stimulation of VO(2) was produced by 0.2 microM PPAHV (delta VO(2), 0.83+/-0.06 micromol g(-1) h(-1)) and was accompanied by mild vasoconstriction (increase in PP; 8.0+/-1.1 mmHg). The highest concentration of PPAHV tested (2 microM) caused inhibition of VO(2) (delta VO(2), -2.73+/-0.51 micromol g(-1) h(-1)) and strong vasoconstriction (delta PP, 42.0+/-1.2 mmHg). 3. Capsazepine (10 microM) caused a parallel shift to the right of both VO(2) and PP concentration-response curves for PPAHV (pK(b)=5.00), indicative of competitive binding to vanilloid receptors. 4. The stimulation of VO(2) produced by 0.2 microM PPAHV decreased, but was not completely abolished, after repeated infusion of PPAHV (change in VO(2), first infusion, 0.66+/-0.18 micromol g(-1) h(-1); sixth infusion, 0.29+/-0. 08 micromol g(-1) h(-1), P<0.05), an acute tachyphylactic response not previously seen with the repeated infusion of other vanilloid analogues. Conversely, the PP response to repeated PPAHV infusion increased (delta PP, first infusion, 5.8+/-0.7 mmHg; sixth infusion, 9.0+/-0.6 mmHg, P<0.05). 5. In conclusion, PPAHV produces vasoconstriction and a biphasic effect on VO(2) in the perfused rat hindlimb very similar to that induced by naturally occurring vanilloids. Both effects are blocked by the competitive antagonist capsazepine. Since, the metabolic response to low concentrations of PPAHV (stimulation of VO(2)) undergoes tachyphylaxis, the present data suggest that PPAHV desensitizes putative vanilloid receptors in the hindlimb.
Assuntos
Capsaicina/análogos & derivados , Membro Posterior/efeitos dos fármacos , Ésteres de Forbol/farmacologia , Animais , Capsaicina/farmacologia , Relação Dose-Resposta a Droga , Membro Posterior/fisiologia , Masculino , Consumo de Oxigênio/efeitos dos fármacos , Perfusão , Ratos , Ratos Wistar , Resistência Vascular/efeitos dos fármacos , Vasoconstrição/efeitos dos fármacosRESUMO
1. The characteristics, localization and regulation of tachykinin receptors in the rat nucleus tractus solitarius (NTS) involved in respiratory control were investigated using a combination of in vivo microinjection and in vitro autoradiographic techniques. 2. Microinjection of receptor-selective tachykinin agonists and antagonists into the NTS of urethane-anaesthetized rats suggests that stimulation of NK1 and NK3 receptors increases tidal volume, whereas NK2 and NK3 receptor activation produces a bradypnoea. 3. Depletion of NK1 receptors in the NTS due to either ageing or acute hypoxia correlates with a markedly reduced respiratory response to substance P. In contrast, chemical ablation of sensory neurons by neonatal capsaicin administration dramatically increases the respiratory response to a variety of NK1, NK2 and NK3 agonists. 4. These studies suggest that all three tachykinin receptors are present in the rat NTS and that these receptors are subject to both acute and chronic regulation.
Assuntos
Receptores de Taquicininas/metabolismo , Núcleo Solitário/metabolismo , Animais , Ratos , Receptores de Taquicininas/efeitos dos fármacos , Receptores de Taquicininas/fisiologia , Núcleo Solitário/efeitos dos fármacos , Núcleo Solitário/fisiologiaRESUMO
1. The respiratory response to microinjection of tachykinins and analogues into the commissural nucleus of the solitary tract (cNTS) of urethane-anaesthetized rats was investigated in the presence and absence of selective tachykinin NK(1), NK(2) and NK(3) antagonists (RP 67580, SR 48968 and SR 142801, respectively). 2. All tachykinins, except for the selective NK(2) agonist, [Nle(10)]-NKA(4-10), increased tidal volume (VT). The rank potency order of naturally-occurring tachykinins was neurokinin A (NKA)> or =substance P (SP)>>NKB, whereas the rank order for selective analogues was senktide> or = septide>> [Sar(9),Met(O(2))(11)]-SP>>[Nle(10)]-NKA(4-10). Septide (NK(1)-selective) and senktide (NK(3)-selective) were 22 fold more potent (pD(2) approximately 12) at stimulating VT than SP (pD(2) approximately 10.5). 3. Tachykinin agonists produced varying degrees of respiratory slowing, independent of changes in VT. At doses producing maximum stimulation of VT, agonists induced either a mild (<10 breaths min(-1) decrease; SP and septide), moderate (10 - 25 breaths min(-1) decrease; NKA, NKB and [Sar(9),Met(O(2)]-SP) or severe ( approximately 40 breaths min(-1) decrease; senktide) bradypnoea. [Nle(10)]-NKA(4-10) produced a dose-dependent bradypnoea without affecting VT. 4. RP 67580 significantly attenuated the VT response to SP (33 pmol) and NKA (10 pmol) but not NKB (100 pmol). In the presence of RP 67580, the mild bradypnoeic response to NKB was significantly enhanced whereas SP and NKA induced a bradyapnea which was not observed in the absence of RP 67580. SR 48968 had no effect on the VT response to SP or NKB, markedly enhanced the VT response to NKA and completely blocked the bradypnoeic response to [Nle(10)]-NKA(4-10). Only SR142801 attenuated the VT response to NKB. 5. The present data suggest that all three tachykinin receptors (NK(1), NK(2) and NK(3)) are present in the cNTS and are involved in the central control of respiration.
Assuntos
Receptores de Taquicininas/agonistas , Receptores de Taquicininas/antagonistas & inibidores , Mecânica Respiratória/efeitos dos fármacos , Núcleo Solitário/efeitos dos fármacos , Taquicininas/farmacologia , Animais , Benzamidas/farmacologia , Indóis/farmacologia , Isoindóis , Masculino , Microinjeções , Neurocinina A/administração & dosagem , Neurocinina A/farmacologia , Neurocinina B/administração & dosagem , Neurocinina B/farmacologia , Antagonistas dos Receptores de Neurocinina-1 , Piperidinas/farmacologia , Ratos , Ratos Wistar , Receptores da Neurocinina-2/antagonistas & inibidores , Receptores da Neurocinina-3/antagonistas & inibidores , Substância P/administração & dosagem , Substância P/farmacologia , Volume de Ventilação Pulmonar/efeitos dos fármacosRESUMO
1. The respiratory response to microinjection of capsaicin and tachykinin receptor agonists into the commissural nucleus of the solitary tract (cNTS) was investigated in adult, urethane-anaesthetized rats which had been pretreated with capsaicin (50 mg kg(-1) s.c.) or vehicle (10% Tween 80, 10% ethanol in saline) as day 2 neonates. 2. Microinjection of capsaicin (1 nmol) into the cNTS of vehicle-pretreated rats, significantly reduced respiratory frequency (59 breaths min(-1), preinjection control, 106 breaths min(-1)) without affecting tidal volume (VT). In capsaicin-pretreated rats, the capsaicin-induced bradypnoea was markedly attenuated (minimum frequency, 88 breaths min(-1); control, 106 breaths min(-1)). 3. In vehicle-pretreated rats, microinjection of substance P (SP, 33 pmol), neurokinin A (NKA, 33 pmol) and NKB (330 pmol), and the selective NK(1) tachykinin receptor agonists, [Sar(9), Met(O(2))(11)]-SP (33 pmol) and septide (10 pmol), increased VT (maxima, 3.60 - 3.93 ml kg(-1)) compared with preinjection control (2.82 ml kg(-1)), without affecting frequency. The selective NK(3) agonist senktide (10 pmol) also increased VT (3.93 ml kg(-1)) which was accompanied by a bradypnoea (-25 breaths min(-1)). The selective NK(2) agonist, [Nle(10)]-NKA(4-10) (330 pmol) increased VT slightly but significantly decreased frequency (-12 breaths min(-1)). In capsaicin-pretreated rats, VT responses to SP and [Sar(9), Met(O(2))(11)]-SP were increased whereas the response to septide was abolished. Both the VT and bradypnoeic responses to senktide and [Nle(10)]-NKA(4-10) were significantly enhanced. 4. These results show that neonatal capsaicin administration markedly reduces the respiratory response to microinjection of capsaicin into the cNTS. The destruction of capsaicin-sensitive afferents appears to sensitize the NTS to SP, NKB, [Sar(9),Met(O(2))(11)]-SP, senktide and [Nle(10)]-NKA(4-10). Moreover, the loss of septide responsiveness in capsaicin-pretreated rats, suggests that 'septide-sensitive' NK(1) receptors may be located on the central terminals of afferent neurons.
Assuntos
Animais Recém-Nascidos/fisiologia , Capsaicina/farmacologia , Mecânica Respiratória/efeitos dos fármacos , Núcleo Solitário/efeitos dos fármacos , Taquicininas/farmacologia , Animais , Masculino , Microinjeções , Neurocinina A/administração & dosagem , Neurocinina A/farmacologia , Neurocinina B/administração & dosagem , Neurocinina B/farmacologia , Fragmentos de Peptídeos/farmacologia , Ratos , Ratos Wistar , Substância P/análogos & derivados , Substância P/farmacologiaRESUMO
1. The respiratory response to microinjection of capsaicin into the commissural nucleus of the solitary tract (cNTS) of urethane-anaesthetized rats was investigated in the absence and presence of the competitive vanilloid (capsaicin) antagonist, capsazepine, and selective tachykinin NK1, NK2 and NK3 antagonists (RP 67580, SR 48968 and SR 142801, respectively). 2. Microinjection of capsaicin reduced respiratory frequency but not tidal volume (VT), leading to an overall reduction in minute ventilation (VE). The effect was dose-dependent between 0.5 and 2 nmol capsaicin. Doses greater than 2 nmol produced apnoea. Tachyphylaxis was observed following repeated injection of capsaicin (1 nmol, 30 min apart). 3. Capsazepine (1 nmol) had no effect on frequency or VT when injected alone but completely blocked the respiratory response to capsaicin (1 nmol). 4. RP 67580 (1 but not 5 nmol) alone depressed frequency and VT slightly. Moreover, RP 67580 appeared to potentiate the bradypnoeic effect of capsaicin. In contrast, SR 48968 and SR 142801 (1 and 5 nmol) alone had no significant effect on respiration. However, both agents significantly attenuated the reduction in frequency produced by capsaicin. 5. In conclusion, microinjection of capsaicin into the cNTS decreases overall ventilation, primarily by reducing frequency. The action of capsaicin appears from the data to be mediated by vanilloid receptors since it is blocked by the competitive vanilloid antagonist capsazepine and is subject to tachyphylaxis. However, since NK2 (SR 48968) and NK3 (SR 142801) receptor antagonists block the actions of capsaicin, we propose that capsaicin acts also by releasing tachykinins from central afferent terminals in the cNTS.
Assuntos
Capsaicina/farmacologia , Receptores de Droga/efeitos dos fármacos , Receptores de Taquicininas/efeitos dos fármacos , Mecânica Respiratória/efeitos dos fármacos , Núcleo Solitário/citologia , Anestesia , Animais , Benzamidas/farmacologia , Capsaicina/administração & dosagem , Capsaicina/análogos & derivados , Capsaicina/antagonistas & inibidores , Relação Dose-Resposta a Droga , Indóis/farmacologia , Isoindóis , Masculino , Microinjeções , Antagonistas dos Receptores de Neurocinina-1 , Piperidinas/farmacologia , Ratos , Ratos Wistar , Receptores de Droga/antagonistas & inibidores , Receptores da Neurocinina-2/antagonistas & inibidores , Receptores da Neurocinina-3/antagonistas & inibidores , Receptores de Taquicininas/antagonistas & inibidores , Núcleo Solitário/efeitos dos fármacos , Volume de Ventilação Pulmonar/efeitos dos fármacosRESUMO
The respiratory response to microinjection of substance P (SP) into the commissural nucleus of the solitary tract (cNTS) and binding of [125I]-Bolton-Hunter SP ([125I]-BHSP) to brain stem NK1 receptors were compared in young and aged rats. Injection of SP (750 pmol) into the cNTS of young rats (2 months) increased tidal volume (VT) but had no effect on respiratory rate (f). In aged rats (19-21 months), injection of SP had no significant effect on f or VT. The NTS of aged rats displayed significantly lower specific [125I]-BHSP binding than young rats, indicating a reduction in the number in NK1 receptors. These findings show that the respiratory response to microinjection of SP into the cNTS of aged rats is severely blunted and that this phenomenon may be due to a decrease in the number of NK1 receptors in the NTS.
Assuntos
Envelhecimento/fisiologia , Receptores da Neurocinina-1/metabolismo , Respiração , Núcleo Solitário/metabolismo , Substância P/farmacologia , Succinimidas/farmacologia , Animais , Autorradiografia , Radioisótopos do Iodo , Masculino , Ensaio Radioligante , Ratos , Ratos Wistar , Núcleo Solitário/químicaRESUMO
Prolonged or repetitive bouts of hypoxia may desensitize the brain stem respiratory centres leading to reduced stimulation of ventilation. We investigated the possible involvement of changes in the sensitivity of the commissural nucleus of the solitary tract (cNTS) to the tachykinin peptide, substance P (SP). Urethane-anaesthetised rats were allowed to breath room air (normoxic) or subjected to four, 30 s bouts of hypoxia (10% O2/90% N2) prior to the injection of SP (750 pmol) into the cNTS. In normoxic rats (n = 5), SP produced a fall in frequency (f, 88+/-4% control) after 4 min and a maximum rise in tidal volume (VT) after 6 min (138+/-10% control) leading to an overall increase in minute ventilation (VE, maximum, 127+/-12% control after 2 min). In rats (n = 5) exposed to four bouts of hypoxia and allowed to recover for 10 min, injection of SP produced a similar fall in f but a delayed and significantly (P < 0.001) reduced VT (maximum after 10 min, 110+/-1% control) and hence, VE response (104+/-3% control). Sixty min after hypoxia, the f, VT and VE responses to SP were identical to those of normoxic rats. These data suggest that hypoxia desensitizes SP receptors in the cNTS and this may partly explain why the respiratory response to hypoxia declines over time.
Assuntos
Hipóxia/fisiopatologia , Respiração/efeitos dos fármacos , Núcleo Solitário/fisiologia , Substância P/farmacologia , Animais , Injeções , Injeções Intraventriculares , Masculino , Ratos , Ratos Wistar , Valores de Referência , Volume de Ventilação Pulmonar/efeitos dos fármacosRESUMO
In perfused rat skeletal muscle (hindlimb), capsaicin either stimulates (submicromolar concentrations) or inhibits (micromolar concentrations) oxygen consumption (VO2). Both VO2 effects are associated with vasoconstriction, evident as an increase in perfusion pressure (PP), under constant flow. We have proposed that these effects are mediated by two vanilloid receptor subtypes: VN1 (stimulation of VO2) and VN2 (inhibition of VO2) (; ). In the present study, the role of capsaicin-sensitive neurons and sensory neuropeptides in the VN1/VN2 receptor actions of capsaicin was investigated. The observed maximum stimulation of VO2 by capsaicin (0.4 microM; DeltaVO2, 1.35 +/- 0.14 micromol g-1 h-1) was accompanied by mild vasoconstriction (DeltaPP, 5.8 +/- 0.6 mm Hg). In contrast, 2 microM capsaicin produced strong inhibition of VO2 (DeltaVO2, -2.25 +/- 0.23 micromol g-1 h-1) with pronounced vasoconstriction (DeltaPP, 28.0 +/- 1.3 mm Hg). VO2 stimulation was significantly inhibited (P <.05) by the selective NK1 receptor antagonist CP-99994 (1 microM) and the NK2 receptor antagonist SR 48968 (1 microM) (by 42% and 51%, respectively), but PP was not altered. Infused SP and neurokinin A (NKA) stimulated VO2 (observed maximum DeltaVO2, 0.52 +/- 0.06 and 0.53 +/- 0.08 micromol g-1 h-1, respectively; EC50 values, 269 +/- 23 and 21.2 +/- 3.0 nM, respectively) and induced mild vasoconstriction (4.30 +/- 0.33 and 6. 75 +/- 1.18 mm Hg, respectively; EC50 values, 352 +/- 25.7 and 25.5 +/- 2.7 nM, respectively). Neurokinin B (NKB) also stimulated VO2 (maximum not determined) and vasoconstriction (maximum DeltaPP, 3.40 +/- 0.25 mm Hg; EC50, 34.4 +/- 5.2 nM). The rank order of potency for the tachykinins in this preparation was NKA > NKB > SP, which suggests stimulation primarily of NK2 receptors. Although infused calcitonin gene-related peptide (CGRP) did not alter hindlimb VO2 or PP, the selective CGRP antagonist CGRP(8-37) markedly potentiated the inhibition of VO2 produced by 1 microM capsaicin (84%) and the maximum capsaicin-induced vasoconstriction (57%), which indicates that endogenously released CGRP may act as a vasodilator. Hindlimbs perfused 1 day after capsaicin pretreatment showed attenuation of capsaicin-induced (0.4 microM) stimulation of VO2 (92%) (P <.05) and vasoconstriction (64%), but this returned to normal after 7 days. The inhibition of VO2 by 1 microM capsaicin was significantly (P <. 05) enhanced 7 and 14 days after pretreatment (66% and 140%, respectively), as was the maximum vasoconstriction (64% and 68%, respectively). These data suggest that capsaicin-sensitive neurons, presumably via release of SP and NKA, are involved in VN1 responses and that capsaicin pretreatment potentiates VN2 responses, either by depletion of CGRP reserves or by upregulation of putative VN2 receptors.
Assuntos
Peptídeo Relacionado com Gene de Calcitonina/fisiologia , Capsaicina/farmacologia , Músculo Esquelético/efeitos dos fármacos , Neurocinina A/fisiologia , Fragmentos de Peptídeos/fisiologia , Substância P/fisiologia , Animais , Benzamidas/farmacologia , Masculino , Piperidinas/farmacologia , Ratos , Ratos Wistar , Receptores da Neurocinina-2/antagonistas & inibidoresRESUMO
This study utilised autoradiography to examine [125I]-Bolton Hunter substance P (BHSP) binding in postmortem human visual cortex. In the primary visual area, layers I-III, IVC and VI exhibited low levels of BHSP binding, while high levels were observed in layers IVB and V. Because cells in layers IVB and V are known to be involved in processing direction-specific stimuli, it is possible that SP plays a role in modulating this visual process.
Assuntos
Receptores da Neurocinina-1/análise , Córtex Visual/química , Adulto , Idoso , Autorradiografia , Humanos , Masculino , Pessoa de Meia-Idade , Ensaio RadioliganteRESUMO
Binding of [125I]-labeled Bolton-Hunter substance P ([125I]-BHSP) to NK1 receptors was investigated in the spinal cord of young (3-4 month) and aged (14-16 month) rats. In homogenates of whole spinal cord, the affinity (equilibrium dissociation constant, approximately 210 pM) and maximum density of [125I]-BHSP binding sites ( approximately 0.25 fmol/mg wet weight) were similar for young and aged rats. Autoradiographic studies revealed a similar distribution of [125I]-BHSP sites in both young and old rats at all spinal levels. Intense binding was observed in the superficial dorsal horn (laminae I-III), grey commissure (lamina X) and thoracic intermediolateral cell column (IML) with lower levels of binding in the deeper dorsal horn (laminae IV-VI) and ventral horn (laminae VII-IX). However, the density of [125I]-BHSP sites was significantly (P<0.05) lower in lamina X of lumbar sections of aged rats compared with young controls. These studies suggest that ageing is associated with a selective loss of NK1 receptors in lamina X of the lumbar spinal cord, although the affinity of NK1 receptors in aged rats is unchanged.
Assuntos
Envelhecimento/metabolismo , Medula Espinal/metabolismo , Substância P/metabolismo , Succinimidas/metabolismo , Animais , Autorradiografia , Sítios de Ligação/fisiologia , Radioisótopos do Iodo , Região Lombossacral , Masculino , Ratos , Ratos Wistar , Receptores da Neurocinina-1/metabolismo , Medula Espinal/crescimento & desenvolvimentoRESUMO
Substance P (SP) is a key neurotransmitter involved in the brain stem integration of carotid body chemoreceptor reflexes. In this study, the characteristics and location of SP receptors in the rat brain stem and their regulation by hypoxia were investigated using homogenate radioligand binding and quantitative autoradiography. Specific binding of [125I] Bolton-Hunter SP (BHSP) to brain stem homogenates was saturable (approximately 0.3 nM) and to a single class of high-affinity sites (K(d), 0.16 nM; maximum density of binding sites, 0.43 fmol/mg wet weight tissue). The order of potency of agonists for inhibition of BHSP binding was SP > [Sar9Met(O2)11]SP >> neurokinin A > septide > neurokinin B >> [Nle10]-neurokinin A(4-10) = senktide, and for nonpeptide antagonists, RP 67580 > CP-96,345 >> RP 68651 = CP-96,344, consistent with binding to NK1 receptors. The effect of single and multiple, 5-min bouts of hypoxia (8.5% O2/91.5% N2) on BHSP binding was investigated using quantitative autoradiography. Binding sites were localized to the lateral, medial and commissural nucleus of the solitary tract (NTS), the hypoglossal nucleus, central gray and the spinal trigeminal tract and nucleus (Sp5 and nSp5, respectively). Five min after a single bout of hypoxia, the density of BHSP binding sites had decreased significantly (P < .05) in the medial NTS (-33%) and lateral NTS (-24%) when compared to normoxic controls. However, the normal receptor complement was restored within 60 min of the hypoxic challenge. In the Sp5, a significant decrease (P < .05) in binding was observed 5 min after hypoxia which was still apparent after 60 min. In contrast, the density of BHSP binding sites in the hypoglossal nucleus decreased slowly and was significantly lower (P < .05) than normoxic controls 60 min after hypoxia. Five min after repetitive hypoxia (3 x 5 min bouts), BHSP binding in the NTS was reduced by more than 40%. Studies in homogenates showed that the affinity of SP for BHSP binding sites was not affected by repetitive hypoxia (K(d)s, normoxic, 0.27 nM; hypoxic, 0.24 nM). These data suggest that afferent input from carotid body chemoreceptors may dynamically regulate NK1 receptors in several brain stem nuclei that are intimately involved in stimulating ventilation during hypoxia, and that the time-course of receptor turnover may differ from region to region in the brain stem. The temporary loss of NK1 receptors in the NTS may partly explain why adequate ventilation is often not maintained during hypoxia.
Assuntos
Hipóxia/metabolismo , Receptores da Neurocinina-1/análise , Centro Respiratório/química , Animais , Masculino , Ratos , Ratos Wistar , Respiração , Substância P/metabolismo , Succinimidas/metabolismoRESUMO
Previous studies with the vanilloid spice principle capsaicin have demonstrated a biphasic VO2 response, with vasoconstriction, in the perfused rat hindlimb that has led to suggestions of vanilloid receptor subtypes (VN1/VN2) in this preparation (1). In the present study, the known competitive vanilloid antagonist capsazepine inhibited the above capsaicin-mediated effects in a manner that was indicative of binding at specific vanilloid recognition sites. Low concentrations of capsazepine selectively inhibited the increased VO2 produced by the putative VN1 receptor at submicromolar concentrations of capsaicin, while the inhibition of VO2 produced by high concentrations of capsaicin (putative VN2) was enhanced. These observations, showing different susceptibilities to blockade by capsazepine, further support the presence of two vanilloid receptor subtypes in the rat hindlimb. Schild plots of the data yielded variable slopes that approach unity at greater responses to capsaicin (mean KB = 8.44 +/- 2.08 microM and 7.28 +/- 0.78 microM for VO2 and perfusion pressure curves, respectively). Low concentrations of the capsaicin antagonist ruthenium red selectively blocked the putative VN2 receptor-mediated effects produced by high concentrations of capsaicin. The noncompetitive nature of this inhibitor suggests an operation through separate receptor-coupled ion channel complexes at high and low concentrations of the vanilloid. Tetrodotoxin failed to attenuate any changes produced by capsaicin, suggesting that the mechanism of action of capsaicin in the rat hindlimb may differ from other tissues.
Assuntos
Capsaicina/análogos & derivados , Capsaicina/farmacologia , Músculos/efeitos dos fármacos , Oxigênio/metabolismo , Receptores de Droga/efeitos dos fármacos , Rutênio Vermelho/farmacologia , Animais , Relação Dose-Resposta a Droga , Membro Posterior/efeitos dos fármacos , Masculino , Ratos , Ratos WistarRESUMO
Tachykinin receptors in guinea-pig airways were examined using radioligand binding techniques in lung homogenates, and using isolated bronchial segments. Binding of the NK1 selective radioligand 125I-labelled Bolton-Hunter [Sar9,Met(O2)11]substance P ([125I]BHSarSP) was saturable and of high affinity (KD, 0.26 nM). The rank potency order of competitors for [125I]BHSarSP binding was [Pro9]SP > CP 96345 >> septide > [pGlu6]SP(6-11) > RP 67580 > or = [DPro9,t beta Pro10(phi),Trp11]SP > [DPro9,t beta Pro10(CH2 phi),Trp11]physalaemin > or = GR82334 > or = 127I Bolton-Hunter neurokinin A (BHNKA). Septide had higher affinity than expected, and it was the only ligand to bind to two sites. Agonists interacting with NK2 receptors were more potent contractile agents than NK1 receptor agonists. Responses to BHNKA (pD2 8.4) were antagonized by MDL 29913 and MEN 10207, with pKB values 6.42 and 6.79, and also by SR 48968 and GR 94800, although this was not dose dependent. This agonist was also weakly inhibited by CP 96345 and RP 67580. These data demonstrate that BHNKA can interact with both NK1 and NK2 receptors. There was no relationship between the binding affinity of NK1 ligands in lung homogenates, with GR 82334 being notably weak, and their agonist or antagonist potency in bronchial smooth muscle.
Assuntos
Pulmão/química , Receptores da Neurocinina-1/análise , Receptores da Neurocinina-2/análise , Animais , Compostos de Bifenilo/farmacologia , Feminino , Cobaias , Técnicas In Vitro , Masculino , Fisalemina/análogos & derivados , Fisalemina/farmacologia , Ensaio Radioligante , Substância P/análogos & derivados , Substância P/metabolismoRESUMO
Previous work has demonstrated contraceptive steroid-induced hypertension in rats. Here, we examined the relationship between steroid-induced hypertension and components of the renin-angiotensin system. Female Sprague-Dawley rats were injected s.c. with 0.2 micrograms ethynyloestradiol, 2.0 micrograms levonorgestrel, a combination of both or vehicle, six days per week. A second group of rats received 2.0 micrograms enalapril maleate, enalapril plus ethynyloestradiol or levonorgestrel, or vehicle. Systolic blood pressure increased with both ethynyloestradiol. (6 weeks, +17 mmHg; 12 weeks, +32 mmHg) and levonorgestrel (6 weeks, +24 mmHg) treatment. This effect of levonorgestrel was attenuated by co-administration of enalapril, which also reversed the hypertension seen with ethynyloestradiol. Ethynyloestradiol, but not levonorgestrel treatment caused a significant increase in plasma renin concentration, plasma renin activity, and plasma angiotensin II at both 6 and 12 weeks. Plasma renin substrate was increased by ethynyloestradiol at 3, 6 and 12 weeks, prior to the observed increase in systolic blood pressure. Combined steroid treatment had less pronounced effects. Enalapril alone or in combination with ethynyloestradiol decreased plasma renin concentration, activity and angiotensin II, and in combination with levonorgestrel decreased plasma renin concentration, substrate and activity (6 weeks only) but not angiotensin II. The data indicate a positive relationship between hypertension and the renin-angiotensin system with ethynyloestradiol, but not levonorgestrel treatment in rats.