RESUMO
The carbonic anhydrase inhibitor dorzolamide can induce relaxation of retinal arterioles with a consequent increase in blood flow and oxygenation of the retina. It has been shown that the mechanisms underlying this relaxation are independent of extracellular acidosis and CO2. The purpose of the present study was to investigate the possible involvement of nitric oxide (NO) and intracellular acidosis in dorzolamide-induced relaxation of retinal arterioles. Porcine retinal arterioles were mounted in a wire myograph and dorzolamide induced relaxation was studied after 1) the addition of the NO synthase inhibitor l-NAME (3 × 10(-4) M) or the guanylyl cyclase inhibitor ODQ (3 × 10(-6) M), and 2) after loading the smooth muscle cells with the pH sensitive fluorophore SNARF-1-AM and studying changes in vascular tone and intracellular fluorescence after the induction of hypoxia, addition of lactate (10(-2) M), and extracellular acidification (pH = 7.0) alone and in the presence of dorzolamide (10(-3) M). Dorzolamide significantly relaxed retinal arterioles (p < 0.03), and the effect was significantly higher in the presence of perivascular tissue than in isolated vessels at the highest concentration (p < 0.01). In the presence of perivascular tissue dorzolamide-induced relaxation could be reduced by NO inhibition (p < 0.02). Dorzolamide increased intracellular acidification (p < 0.02) during extracellular acidosis, but there was no relation between relaxation and intracellular acidosis. In conclusion, dorzolamide-induced vasorelaxation depends on NO and the perivascular retinal tissue, but is independent of acidification in the extracellular and the intracellular space of retinal vascular smooth muscle cells. Other factors than NO and acidification are involved in dorzolamide-induced relaxation of retinal arterioles.
Assuntos
Acidose/metabolismo , Inibidores da Anidrase Carbônica/farmacologia , Músculo Liso Vascular/efeitos dos fármacos , Óxido Nítrico/metabolismo , Artéria Retiniana/fisiologia , Sulfonamidas/farmacologia , Tiofenos/farmacologia , Vasodilatação/fisiologia , Animais , Arteríolas/efeitos dos fármacos , Benzopiranos/metabolismo , Bradicinina/farmacologia , Endotélio Vascular/fisiologia , Inibidores Enzimáticos/farmacologia , Corantes Fluorescentes/metabolismo , Concentração de Íons de Hidrogênio , Lactatos/farmacologia , Músculo Liso Vascular/metabolismo , Miografia , NG-Nitroarginina Metil Éster/farmacologia , Oxidiazóis/farmacologia , Quinoxalinas/farmacologia , SuínosRESUMO
AIM: Type 2 diabetes is associated with stroke and cardiac dysfunction. We therefore investigated isolated middle cerebral arteries and coronary septal arteries from the diabetic Goto-Kakizaki (GK) rat model of nonobese type 2 diabetes. METHODS: Myogenic tone and agonist-induced responses were investigated under isobaric conditions with simultaneous recording of [Ca2+]i. Rho-kinase and NO pathways were investigated using specific pharmacological tools. RESULTS: Arteries from GK rats developed less tone at pressures from 20 to 100 mm Hg than arteries from control Wistar (CW) rats while [Ca2+]i was similar. Blocking the Rho-kinase pathway decreased the pressure-induced development of tone and after blockade no difference in myogenic tone between arteries from GK and CW rats was seen. Cerebral arteries had similar tone to a maximal concentration of U46619 (GK: 35.5±2% vs. CW: 31.6±5%), while coronary arteries from GK rats developed less tone than arteries from CW rats (12±3 vs. 26.1±3%). Endothelium-dependent vasodilation to A23187 (cerebral) and to acetylcholine (coronary) was not different between arteries from GK and CW rats. CONCLUSION: Our data suggest that in resistance arteries from the brain and the heart of GK rats the myogenic tone is decreased due to impaired calcium sensitivity likely due to a defective Rho-kinase pathway.
Assuntos
Vasos Coronários/fisiopatologia , Diabetes Mellitus Tipo 2/fisiopatologia , Artéria Cerebral Média/fisiopatologia , Ácido 15-Hidroxi-11 alfa,9 alfa-(epoximetano)prosta-5,13-dienoico/farmacologia , Amidas/farmacologia , Animais , Glicemia/análise , Cálcio/metabolismo , Modelos Animais de Doenças , Masculino , Artéria Cerebral Média/patologia , Óxido Nítrico/fisiologia , Proteína Quinase C/fisiologia , Piridinas/farmacologia , Ratos , Ratos Wistar , Quinases Associadas a rho/fisiologiaRESUMO
Altered pH-regulatory ion transport is characteristic of many cancers; however, the mechanisms and consequences are poorly understood. Here, we investigate how a truncated, constitutively active ErbB2 receptor (DeltaNErbB2) common in breast cancer impacts on the Na(+)/H(+)-exchanger NHE1 and the Na(+),HCO(3)(-)-cotransporter NBCn1 in MCF-7 human breast cancer cells and address the roles of these transporters in chemotherapy resistance. Upon DeltaNErbB2 expression, mRNA and protein levels of NBCn1, yet not of NHE1, increased several-fold, and the localization of both transporters was altered paralleling extensive morphological changes. The rate of pH(i) recovery after acid loading increased by 50% upon DeltaNErbB2 expression. Knockdown and pharmacological inhibition confirmed the involvement of both NHE1 and NBCn1 in acid extrusion. NHE1 inhibition or knockdown sensitized DeltaNErbB2-expressing cells to cisplatin-induced programmed cell death (PCD) in a caspase-, cathepsin-, and reactive oxygen species-dependent manner. NHE1 inhibition augmented cisplatin-induced caspase activity and lysosomal membrane permeability followed by cysteine cathepsin release. In contrast, NBCn1 inhibition attenuated cathepsin release and had no net effect on viability. These findings warrant studies of NHE1 as a potential target in breast cancer and demonstrate that in spite of their similar transport functions, NHE1 and NBCn1 serve different functions in MCF-7 cells.
Assuntos
Equilíbrio Ácido-Base/genética , Neoplasias da Mama/genética , Proteínas de Transporte de Cátions/fisiologia , Resistencia a Medicamentos Antineoplásicos/genética , Receptor ErbB-2/genética , Simportadores de Sódio-Bicarbonato/fisiologia , Trocadores de Sódio-Hidrogênio/fisiologia , Antineoplásicos/uso terapêutico , Transporte Biológico/genética , Transporte Biológico/fisiologia , Neoplasias da Mama/tratamento farmacológico , Neoplasias da Mama/metabolismo , Neoplasias da Mama/patologia , Catepsinas/metabolismo , Proteínas de Transporte de Cátions/antagonistas & inibidores , Proteínas de Transporte de Cátions/genética , Proteínas de Transporte de Cátions/metabolismo , Linhagem Celular Tumoral , Feminino , Regulação Neoplásica da Expressão Gênica/efeitos dos fármacos , Humanos , Concentração de Íons de Hidrogênio , Membranas Intracelulares/metabolismo , Proteínas Mutantes/genética , Proteínas Mutantes/metabolismo , Estrutura Terciária de Proteína/genética , RNA Interferente Pequeno/farmacologia , Receptor ErbB-2/química , Receptor ErbB-2/metabolismo , Simportadores de Sódio-Bicarbonato/antagonistas & inibidores , Simportadores de Sódio-Bicarbonato/genética , Simportadores de Sódio-Bicarbonato/metabolismo , Trocador 1 de Sódio-Hidrogênio , Trocadores de Sódio-Hidrogênio/antagonistas & inibidores , Trocadores de Sódio-Hidrogênio/genética , Trocadores de Sódio-Hidrogênio/metabolismoRESUMO
How blood flow and pressure to the giraffe's brain are regulated when drinking remains debated. We measured simultaneous blood flow, pressure, and cross-sectional area in the carotid artery and jugular vein of five anesthetized and spontaneously breathing giraffes. The giraffes were suspended in the upright position so that we could lower the head. In the upright position, mean arterial pressure (MAP) was 193 +/- 11 mmHg (mean +/- SE), carotid flow was 0.7 +/- 0.2 l/min, and carotid cross-sectional area was 0.85 +/- 0.04 cm(2). Central venous pressure (CVP) was 4 +/- 2 mmHg, jugular flow was 0.7 +/- 0.2 l/min, and jugular cross-sectional area was 0.14 +/- 0.04 cm(2) (n = 4). Carotid arterial and jugular venous pressures at head level were 118 +/- 9 and -7 +/- 4 mmHg, respectively. When the head was lowered, MAP decreased to 131 +/- 13 mmHg, while carotid cross-sectional area and flow remained unchanged. Cardiac output was reduced by 30%, CVP decreased to -1 +/- 2 mmHg (P < 0.01), and jugular flow ceased as the jugular cross-sectional area increased to 3.2 +/- 0.6 cm(2) (P < 0.01), corresponding to accumulation of approximately 1.2 l of blood in the veins. When the head was raised, the jugular veins collapsed and blood was returned to the central circulation, and CVP and cardiac output were restored. The results demonstrate that in the upright-positioned, anesthetized giraffe cerebral blood flow is governed by arterial pressure without support of a siphon mechanism and that when the head is lowered, blood accumulates in the vein, affecting MAP.
Assuntos
Anestesia Geral , Pressão Sanguínea , Circulação Cerebrovascular , Movimentos da Cabeça , Veias Jugulares/fisiologia , Postura , Ruminantes/fisiologia , Animais , Débito Cardíaco , Artérias Carótidas/diagnóstico por imagem , Artérias Carótidas/fisiologia , Pressão Venosa Central , Gravitação , Veias Jugulares/diagnóstico por imagem , Masculino , Fluxo Sanguíneo Regional , Telemetria , Ultrassonografia DopplerRESUMO
Recently, we showed that rebaudioside A potently stimulates the insulin secretion from isolated mouse islets in a dose-, glucose- and Ca(2+)-dependent manner. Little is known about the mechanisms underlying the insulinotropic action of rebaudioside A. The aim of this study was to define the signalling system by which, rebaudioside A acts. Isolated mouse islets were used in the cAMP[(125)I] scintillation proximity assay to measure total cAMP level, and in a luminometric method to measure intracellular ATP and ADP concentrations. Conventional and permeabilized whole-cell configuration of the patch-clamp technique was used to verify the effect of rebaudioside A on ATP-sensitive K(+)-channels from dispersed single beta cells from isolated mouse islets. Insulin was measured by radioimmunoassay from insulinoma MIN6 cells. In the presence of 16.7 mM glucose, the addition of the maximally effective concentration of rebaudioside A (10(-9) M) increased the ATP/ADP ratio significantly, while it did not change the intracellular cAMP level. Rebaudioside A (10(-9) M) and stevioside (10(-6) M) reduced the ATP-sensitive potassium channel (K(ATP)) conductance in a glucose-dependent manner. Moreover, rebaudioside A stimulated the insulin secretion from MIN6 cells in a dose- and glucose-dependent manner. In conclusion, the insulinotropic effect of rebaudioside A is mediated via inhibition of ATP-sensitive K(+)-channels and requires the presence of high glucose. The inhibition of ATP-sensitive K(+)-channels is probably induced by changes in the ATP/ADP ratio. The results indicate that rebaudioside A may offer a distinct therapeutic advantage over sulphonylureas because of less risk of causing hypoglycaemia.
Assuntos
Diterpenos do Tipo Caurano/farmacologia , Glucose/farmacologia , Células Secretoras de Insulina/efeitos dos fármacos , Insulina/metabolismo , Canais KATP/metabolismo , Bloqueadores dos Canais de Potássio/farmacologia , Difosfato de Adenosina/metabolismo , Trifosfato de Adenosina/metabolismo , Análise de Variância , Animais , Linhagem Celular , AMP Cíclico/metabolismo , Feminino , Glucosídeos/farmacologia , Glibureto/farmacologia , Secreção de Insulina , Células Secretoras de Insulina/metabolismo , Camundongos , Técnicas de Patch-Clamp , Estimulação QuímicaRESUMO
AIMS: The Na,K-ATPase is involved in a large number of regulatory activities including cSrc-dependent signalling. Upon inhibition of the Na,K-ATPase with ouabain, cSrc activation is shown to occur in many cell types. This study tests the hypothesis that acute potentiation of agonist-induced contraction by ouabain is mediated through Na,K-ATPase-cSrc signalling-dependent sensitization of vascular smooth muscle cells to Ca2+ . METHODS: Agonist-induced rat mesenteric small artery contraction was examined in vitro under isometric conditions and in vivo in anaesthetized rats. Arterial wall tension and [Ca2+ ]i in vascular smooth muscle cells were measured simultaneously. Changes in cSrc and myosin phosphatase targeting protein 1 (MYPT1) phosphorylation were analysed by Western blot. Protein expression was examined with immunohistochemistry. The α1 and α2 isoforms of the Na,K-ATPase were transiently downregulated by siRNA transfection in vivo. RESULTS: Ten micromolar ouabain, but not digoxin, potentiated contraction to noradrenaline. This effect was not endothelium-dependent. Ouabain sensitized smooth muscle cells to Ca2+ , and this was associated with increased phosphorylation of cSrc and MYPT1. Inhibition of tyrosine kinase by genistein, PP2 or pNaKtide abolished the potentiating effect of ouabain on arterial contraction and Ca2+ sensitization. Downregulation of the Na,K-ATPase α2 isoform made arterial contraction insensitive to ouabain and tyrosine kinase inhibition. CONCLUSION: Data suggest that micromolar ouabain potentiates agonist-induced contraction of rat mesenteric small artery via Na,K-ATPase-dependent cSrc activation, which increases Ca2+ sensitization of vascular smooth muscle cells by MYPT1 phosphorylation. This mechanism may be critical for acute control of vascular tone.
Assuntos
Sinalização do Cálcio , Artérias Mesentéricas/enzimologia , ATPase Trocadora de Sódio-Potássio/metabolismo , Vasoconstrição , Quinases da Família src/metabolismo , Animais , Sinalização do Cálcio/efeitos dos fármacos , Endotélio Vascular/efeitos dos fármacos , Endotélio Vascular/enzimologia , Inibidores Enzimáticos/farmacologia , Masculino , Artérias Mesentéricas/efeitos dos fármacos , Músculo Liso Vascular/efeitos dos fármacos , Músculo Liso Vascular/enzimologia , Fosforilação , Proteína Fosfatase 1/metabolismo , Interferência de RNA , RNA Interferente Pequeno/genética , RNA Interferente Pequeno/metabolismo , Ratos Wistar , ATPase Trocadora de Sódio-Potássio/antagonistas & inibidores , ATPase Trocadora de Sódio-Potássio/genética , Vasoconstrição/efeitos dos fármacos , Vasoconstritores/farmacologia , Quinases da Família src/antagonistas & inibidoresRESUMO
Vasomotion is the regular variation in tone of arteries. In our study, we suggest a model for the initiation of vasomotion. We suggest that intermittent release of Ca(2+) from the sarcoplasmic reticulum (SR, cytosolic oscillator), which is initially unsynchronized between the vascular smooth muscle cells, becomes synchronized to initiate vasomotion. The synchronization is achieved by an ion current over the cell membrane, which is activated by the oscillating Ca(2+) release. This current results in an oscillating membrane potential, which synchronizes the SR in the vessel wall and starts vasomotion. Therefore, the pacemaker of the vascular wall can be envisaged as a diffuse array of individual cytosolic oscillators that become entrained by a reciprocal interaction with the cell membrane. The model is supported by experimental data. Confocal [Ca(2+)](i) imaging and isometric force development in isolated rat resistance arteries showed that low norepinephrine concentrations induced SR-dependent unsynchronized waves of Ca(2+) in the vascular smooth muscle. In the presence of the endothelium, the waves converted to global synchronized oscillations of [Ca(2+)](i) after some time, and vasomotion appeared. Synchronization was also seen in the absence of endothelium if 8-bromo-cGMP was added to the bath. Using the patch-clamp technique and microelectrodes, we showed that Ca(2+) release can activate an inward current in isolated smooth muscle cells from the arteries and cause depolarization. These electrophysiological effects of Ca(2+) release were cGMP dependent, which is consistent with the possibility that they are important for the cGMP-dependent synchronization. Further support for the model is the observation that a short-lasting current pulse can initiate vasomotion in an unsynchronized artery as expected from the model.
Assuntos
Modelos Cardiovasculares , Vasoconstrição/fisiologia , Sistema Vasomotor/fisiologia , Animais , Relógios Biológicos/efeitos dos fármacos , Relógios Biológicos/fisiologia , Cafeína/farmacologia , Sinalização do Cálcio , GMP Cíclico/análogos & derivados , GMP Cíclico/farmacologia , Corantes Fluorescentes , Técnicas In Vitro , Contração Isométrica/efeitos dos fármacos , Contração Isométrica/fisiologia , Masculino , Potenciais da Membrana/efeitos dos fármacos , Potenciais da Membrana/fisiologia , Artérias Mesentéricas/efeitos dos fármacos , Artérias Mesentéricas/metabolismo , Microeletrodos , Músculo Liso Vascular/efeitos dos fármacos , Músculo Liso Vascular/metabolismo , Norepinefrina/farmacologia , Técnicas de Patch-Clamp , Inibidores de Fosfodiesterase/farmacologia , Ratos , Ratos Wistar , Resistência Vascular/fisiologia , Vasoconstrição/efeitos dos fármacos , Vasoconstritores/farmacologia , Sistema Vasomotor/efeitos dos fármacosRESUMO
When isolated rat mesenteric small arteries were submitted to 2 s of sonication, a nucleoside triphosphatase activity was released to the medium, mainly from the plasma membrane of the vascular smooth muscle cells. The activity was kinetically characterized: It hydrolysed ATP, UTP and GTP with the same substrate affinity and the same specific activity. CaATP, as well as MgATP were substrates for the enzyme with an apparent Km in the micromolar range. ATPase inhibitors: ouabain, vanadate, AlF4-, oligomycin and N-ethylmaleimide were without effect on the hydrolytic activity. Among other modifiers tested only N,N'-dicyclohexylcarbodiimide caused significant (greater than 30%) inhibition. In the presence of micromolecular concentrations of Ca2+ and Mg2+, small (less than 20 mM) concentrations of Na+, K+, Rb+, Cs+ and choline+, irrespective of the nature of the anion, activated the hydrolysis with an equilibrium ordered pattern, but concentrations of monovalent cation salts above 20 mM decreased the hydrolysis rate. No activation by monovalent cation salts was seen at millimolar concentrations of divalent cations and substrate. On the basis of the results a standard mixture is proposed, which allows a sensitive assay of the specific enzyme activity.
Assuntos
Cálcio/metabolismo , Magnésio/metabolismo , Artérias Mesentéricas/enzimologia , Monoéster Fosfórico Hidrolases/química , Adenosina Trifosfatases/antagonistas & inibidores , Trifosfato de Adenosina/metabolismo , Animais , Cátions , Cátions Bivalentes , Guanosina Trifosfato/metabolismo , Hidrólise , Masculino , Artérias Mesentéricas/ultraestrutura , Microscopia Eletrônica , Nucleosídeo-Trifosfatase , Ratos , Ratos Endogâmicos , Especificidade por Substrato , Uridina Trifosfato/metabolismoRESUMO
Segments of isolated intact rat mesenteric small arteries were incubated in physiological bicarbonate buffer in the presence of nano- to millimolar concentrations of ATP. ATP was hydrolysed, and when the vessel was transferred from one incubation to another, the enzyme activity was transferred with the vessel, consistent with the presence of an ecto-ATPase. The substrate, ATP, was shown to induce a modification of the hydrolytic activity which occurred the more rapidly the higher the concentration of ATP. The modified system hydrolysed ATP with a decreased substrate affinity. As the substrate induced a modification of the hydrolytic activity, steady-state velocity measurements for determination of kinetic parameters could not be obtained. Nevertheless, it was possible to compare the modification caused by ATP and UTP, and to compare the hydrolysis rates measured with [32P]ATP, [32P]UTP and [32P]GTP. It was concluded that the hydrolytic activity of the vessels did not distinguish between the nucleoside triphosphates (NTPs). In a histidine buffer, the activity was shown to be activated by micromolar concentrations of either Ca2+ or Mg2+, and not to be influenced by inhibitors of P-type, F-type and V-type ATPases. Functional removal of the endothelium before assay did not reduce the measured NTP hydrolysis. At millimolar concentrations of trinucleotide the hydrolysis rate was 10-15 mumol per min per gram of tissue or 0.11-0.17 mumol per min per 10(6) vascular smooth muscle cells. This value is equivalent to the maximal velocity obtained for the Ca2+ or Mg(2+)-dependent NTPase released to the medium upon 2 s of sonication of the vessels (Plesner, L., Juul, B., Skriver, E. and Aalkjaer, C. (1991) Biochim. Biophys. Acta 1067, 191-200). Comparing the characteristics of the released NTPase to the characteristics of the activity of the intact vessel, they showed a strong resemblance, but the substrate-induced modification of the enzyme was seen only in the intact preparation.
Assuntos
Artérias Mesentéricas/metabolismo , Nucleotídeos/metabolismo , Adenosina Trifosfatases/antagonistas & inibidores , Adenosina Trifosfatases/metabolismo , Trifosfato de Adenosina/metabolismo , Animais , Cátions Bivalentes , Guanosina Trifosfato/metabolismo , Hidrólise , Técnicas In Vitro , Cinética , Masculino , Artérias Mesentéricas/enzimologia , Ratos , Ratos Endogâmicos , Especificidade por Substrato , Uridina Trifosfato/metabolismoRESUMO
OBJECTIVES: This study investigated the role of angiotensin receptor subtype 1 (AT1) and angiotensin receptor subtype 2 (AT2) in the regulation of Na+-H+ exchanger (NHE) and Na+-HCO3 symporter (NBC) in the infarcted myocardium. BACKGROUND: The cardiac renin-angiotensin system is activated after myocardial infarction (MI), and both angiotensin AT1 and AT2 receptors are upregulated in the myocardium. METHODS: Na+-H+ exchanger isoform-1 and NBC-1 gene expression were determined by reverse transcription polymerase chain reaction and Northern blot analysis; protein levels by Western blot analysis; and activity by measurement of H+ transport in left ventricular (LV) free wall, interventricular septum (IS) and right ventricle (RV) after induction of MI. Rats were treated with placebo, the angiotensin-converting enzyme inhibitor ramipril (1 mg/kg/day), the AT1 receptor antagonist valsartan (10 mg/kg/day) or the AT2 receptor antagonist PD 123319 (30 mg/kg/day). Treatment was started seven days before surgery. RESULTS: Na+-H+ exchanger isoform-1 and NBC-1 messenger RNA (mRNA) expression and protein levels were increased twofold in the LV free wall after MI, whereas no changes were observed in the IS and RV. Na+-dependent H+ flux was increased in the LV free wall. Ramipril inhibited mRNA and protein upregulation of both transporters. Valsartan inhibited the upregulation of NHE-1 mRNA and protein but had no effect on NBC-1 mRNA expression and translation. In contrast, PD 123319 abolished the upregulation of NBC-1 mRNA and protein but had no effect on NHE-1 upregulation. Ramipril and valsartan prevented post-MI increase in NHE-1 activity, whereas ramipril and PD 123319 decreased NBC-1 activity. CONCLUSIONS: Angiotensin II via its AT1 and AT2 receptors differentially controls transcriptional and translational regulation as well as the activity of NHE-1 and NBC-1 in the ischemic myocardium and contributes to the control of pH regulation in cardiac tissue.
Assuntos
Angiotensina II , Angiotensina I , Bicarbonatos/metabolismo , Proteínas de Transporte/metabolismo , Infarto do Miocárdio/metabolismo , Miocárdio/metabolismo , Receptores de Angiotensina/fisiologia , Trocadores de Sódio-Hidrogênio/metabolismo , Animais , Northern Blotting , Masculino , RNA Mensageiro/metabolismo , Distribuição Aleatória , Ratos , Ratos Wistar , Sistema Renina-Angiotensina/fisiologia , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Simportadores de Sódio-BicarbonatoRESUMO
BACKGROUND: The tallest animal on earth, the giraffe (Giraffa camelopardalis) is endowed with a mean arterial blood pressure (MAP) twice that of other mammals. The kidneys reside at heart level and show no sign of hypertension-related damage. We hypothesized that a species-specific evolutionary adaption in the giraffe kidney allows normal for size renal haemodynamics and glomerular filtration rate (GFR) despite a MAP double that of other mammals. METHODS: Fourteen anaesthetized giraffes were instrumented with vascular and bladder catheters to measure glomerular filtration rate (GFR) and effective renal plasma flow (ERPF). Renal interstitial hydrostatic pressure (RIHP) was assessed by inserting a needle into the medullary parenchyma. Doppler ultrasound measurements provided renal artery resistive index (RI). Hormone concentrations as well as biomechanical, structural and histological characteristics of vascular and renal tissues were determined. RESULTS: GFR averaged 342 ± 99 mL min(-1) and ERPF 1252 ± 305 mL min(-1) . RIHP varied between 45 and 140 mmHg. Renal pelvic pressure was 39 ± 2 mmHg and renal venous pressure 32 ± 4 mmHg. A valve-like structure at the junction of the renal and vena cava generated a pressure drop of 12 ± 2 mmHg. RI was 0.27. The renal capsule was durable with a calculated burst pressure of 600 mmHg. Plasma renin and AngII were 2.6 ± 0.5 mIU L(-1) and 9.1 ± 1.5 pg mL(-1) respectively. CONCLUSION: In giraffes, GFR, ERPF and RI appear much lower than expected based on body mass. A strong renal capsule supports a RIHP, which is >10-fold that of other mammals effectively reducing the net filtration pressure and protecting against the high MAP.
Assuntos
Pressão Arterial/fisiologia , Girafas/fisiologia , Hemodinâmica/fisiologia , Rim/fisiologia , Animais , Feminino , Taxa de Filtração Glomerular , Rim/irrigação sanguínea , MasculinoRESUMO
The possible role of Na+ in the development of hypertension in rats was explored in measurements of intracellular Na+, 22Na efflux, and 3H-ouabain binding sites in resistance vessels and skeletal muscles. In resistance vessels obtained from 13-week-old spontaneously hypertensive rats (SHR) or age-matched Wistar-Kyoto rats (WKY), (Na)i, total or ouabain-resistant 22Na efflux, and the concentration of 3H-ouabain binding sites showed no significant differences. Soleus muscles obtained from 6-week-old and 13-week-old SHR contained 5 to 11% more 3H-ouabain binding sites than those of WKY. The small difference in ouabain binding probably was related more to variations in growth rate and strain than to the hypertension. In SHR and WKY the Na+ and K+ contents of gastrocnemius muscles were almost identical at 6 and 13 weeks of age. By contrast, in Wistar rats in which the (Na)i of skeletal muscle was increased sixfold by K+ depletion, the systolic blood pressure was decreased by 10%. The K+ depletion was associated with a 35 to 55% decrease in the concentration of 3H-ouabain binding sites in both resistance vessels and skeletal muscles. The results provide no support for any simple cause-effect relationships between either elevated (Na)i or altered concentration of 3H-ouabain binding sites and hypertension in SHR.
Assuntos
Hipertensão/metabolismo , Músculo Liso Vascular/metabolismo , Músculos/metabolismo , Ouabaína/metabolismo , Potássio/metabolismo , Sódio/metabolismo , Animais , Arteríolas/metabolismo , Pressão Sanguínea , Feminino , Masculino , Ratos , Ratos Endogâmicos SHRRESUMO
In order to investigate whether functional and morphological changes are present in the resistance vasculature before hypertension is established, isolated subcutaneous resistance vessels were studied from 21 young normotensive subjects with a family history of hypertension and 22 controls matched for age, sex, and weight. The vessels from the offspring of hypertensive patients displayed no morphological changes or differences in reactivity or sensitivity to the vasoconstrictor agonists norepinephrine, angiotensin II, serotonin, and vasopressin. In the presence of cocaine, however, vessels from offspring showed a significantly greater shift in sensitivity to norepinephrine, a phenomenon also observed in previous studies of vessels from hypertensive patients. The results suggest that in essential hypertension, while morphological and functional abnormalities of the resistance vasculature may develop as the blood pressure rises, a defect in neuroeffector activity is present before hypertension is established and may be a manifestation of abnormal sympathetic nervous system activity underlying the disease.
Assuntos
Vasos Sanguíneos/fisiopatologia , Hipertensão/fisiopatologia , Resistência Vascular , Adulto , Vasos Sanguíneos/metabolismo , Vasos Sanguíneos/patologia , Cálcio/metabolismo , Cocaína/farmacologia , Resistência a Medicamentos , Humanos , Hipertensão/genética , Hipertensão/metabolismo , Hipertensão/patologia , Técnicas In Vitro , Masculino , Norepinefrina/farmacologiaRESUMO
In a double-blind randomized trial, the effects of treatment with an angiotensin-converting enzyme (ACE) inhibitor (perindopril) and a beta-blocker (atenolol) on small artery structure were compared in previously untreated essential hypertensive patients. Subjects (diastolic blood pressure > or = 100 and < or = 120 mm Hg) were randomly assigned to treatment for 12 months with either perindopril (n = 13, 4 to 8 mg/d) or atenolol (n = 12, 50 to 100 mg/d); the dosage was adjusted upward and in some cases combined (n = 5, perindopril; n = 2, atenolol) with thiazide diuretic to achieve target blood pressure (diastolic blood pressure below 90 mm Hg). Before and at the end of treatment, gluteal biopsies were taken under local anesthetic; from these biopsies, two small arteries were dissected and mounted on a myograph for morphometry. The reduction in blood pressure with atenolol (drop in mean blood pressure 28.4 +/- 1.8 mm Hg) was greater than with perindopril (20.6 +/- 1.8 mm Hg, P < .05). Perindopril treatment caused an increase in small artery diameter (231 +/- 14 to 274 +/- 13 microns, P < .05) and a reduction in the ratio of media thickness to lumen diameter (7.94 +/- 0.65% to 5.96 +/- 0.42%, P < .05), whereas atenolol had no effect (246 +/- 14 to 231 +/- 13 microns and 7.14 +/- 0.47% to 6.79 +/- 0.45%, respectively). The change in small artery morphology caused by perindopril was not accompanied by any change in media cross-sectional area, suggesting that the change was due to "remodeling."(ABSTRACT TRUNCATED AT 250 WORDS)
Assuntos
Inibidores da Enzima Conversora de Angiotensina/uso terapêutico , Artérias/efeitos dos fármacos , Atenolol/uso terapêutico , Hipertensão/tratamento farmacológico , Indóis/uso terapêutico , Adulto , Idoso , Pressão Sanguínea/efeitos dos fármacos , Método Duplo-Cego , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , PerindoprilRESUMO
The purpose of the present study was to determine the cellular basis for the increased ratio of media thickness to lumen diameter (media-lumen ratio) consistently found in the peripheral resistance arteries from patients with essential hypertension using an unbiased stereological principle (the "disector"). Segments of subcutaneous resistance arteries (approximately 200 microns internal diameter) were isolated from gluteal biopsies of skin and subcutaneous fat taken from 16 untreated patients with essential hypertension and 16 age- and sex-matched normotensive control subjects. Measured under standardized conditions (ie, relaxed and under controlled mechanical conditions) on an isometric myograph, vessels from hypertensive patients had a significant (P < .05) reduction in lumen diameter and an increase in media-lumen ratio (P < .05) compared with vessels from normotensive control subjects. These changes were not associated with alterations in the estimated media volume per segment length. After these measurements had been made, the arteries were fixed, serial sectioned, and stained. The volume fraction of smooth muscle cells within the media was estimated by point counting on photomicrographs of the vessels. Using the disector principle, we determined the numerical density (number per unit volume) of smooth muscle cells within the media of each vessel and calculated the average smooth muscle cell volume (1775 +/- 122 [mean +/- SEM] and 1532 +/- 112 microns 3, hypertensive and normotensive, respectively, P > .05) on the basis of these measurements.(ABSTRACT TRUNCATED AT 250 WORDS)
Assuntos
Artérias/patologia , Hipertensão/patologia , Tecido Adiposo/irrigação sanguínea , Pressão Sanguínea , Nádegas , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Valores de Referência , Pele/irrigação sanguínea , Resistência VascularRESUMO
OBJECTIVE: To assess intracellular pH regulation in the presence of bicarbonate in resistance arteries from spontaneously hypertensive rats and Wistar-Kyoto (WKY) rats. METHODS: Intracellular pH was determined in isolated resistance arteries from male adults SHR and WKY rats with the pH-sensitive fluorescent dye bis-carboxyethyl carboxyfluorescein, while the arteries were mounted in a myograph for simultaneous measurements of force. The arteries were acid-loaded using the ammonium chloride technique and the recovery from the acidosis was determined in resting arteries and in arteries activated with 50 mmol/l potassium or arginine vasopressin. This protocol was performed in the presence and in the absence of bicarbonate. RESULTS: In the absence of bicarbonate the intracellular pH was higher in resting arteries from SHR than in those from WKY rats, whereas during activation no significant difference was found. In the presence of bicarbonate no difference in intracellular pH between arteries from SHR and WKY rats could be found. The addition and washout of 15 mmol/l ammonium chloride were associated with large force transients in activated arteries both from SHR and from WKY rats. The proton recovery rate at intracellular pH 6.85 in the absence of bicarbonate was higher in activated arteries from SHR than in those from WKY rats, whereas in resting arteries no significant difference was found. In the presence of bicarbonate no significant difference between SHR and WKY rat arteries was found. CONCLUSION: In the presence of bicarbonate a possible abnormality of the sodium-hydrogen exchange in resistance arteries from SHR is not manifested, because regulation of intracellular pH by bicarbonate-dependent mechanisms can compensate for such an abnormality.
Assuntos
Artérias/metabolismo , Bicarbonatos/farmacologia , Hipertensão/metabolismo , Animais , Concentração de Íons de Hidrogênio , Masculino , Ratos , Ratos Endogâmicos SHR , Ratos Endogâmicos WKY , Trocadores de Sódio-Hidrogênio/fisiologia , Resistência VascularRESUMO
In order to test the hypothesis that the contraction induced in human resistance arterioles by porcine tetradecapeptide renin substrate (TDP) is mediated by enzymes specific to the renin-angiotensin cascade, human resistance vessels from skin and subcutaneous fat were mounted in a myograph and exposed to TDP in the presence and absence of the human renin inhibitor H261, the serine protease inhibitor aprotinin, a polyclonal anti-human renin antibody and captopril. TDP induced a dose-dependent contraction that could be abolished by saralasin. The sensitivity to TDP was significantly attenuated by H261, aprotinin and combinations of captopril with aprotinin and captopril plus aprotinin and H261, as indicated by a significant reduction in pD2 (-log10ED50 [mmol/l]) for TDP. However, captopril alone was ineffective. It was concluded that at physiological pH, porcine TDP induces contraction in human resistance vessels by the action of enzymes not specific to the renin-angiotensin cascade. Whilst a clear inhibitory effect of H261 was demonstrated, a significant comparable inhibition by captopril and a polyclonal renin antibody was not observed. This may reflect the difficulty with which these inhibitors gain access to their intracellularly located substrates.
Assuntos
Renina/metabolismo , Saralasina/farmacologia , Resistência Vascular/efeitos dos fármacos , Estimulação Elétrica , Feminino , Humanos , Masculino , Sistema Renina-Angiotensina/efeitos dos fármacos , Serina Endopeptidases/farmacologia , Vasoconstrição/efeitos dos fármacosRESUMO
OBJECTIVE: To investigate the elastic characteristics of the wall of isolated subcutaneous resistance arteries from patients with essential hypertension, the response of the vessels to endothelium-dependent and -independent vasodilators and the dependence on calcium. METHODS: Subcutaneous resistance arteries were isolated from 16 patients with never-treated essential hypertension and from 16 normotensive controls matched for age and sex. The vessels were mounted in a myograph for isometric force development. The passive elastic characteristics were determined and then the response to acetylcholine, nitroprusside, felodipine, caffeine and calcium (in the presence of noradrenaline and prazosin or yohimbine) were determined. RESULTS: Young's elastic modulus as a function of wall stress was similar in the two groups of vessels. The relaxation of vessels from hypertensive and normotensive in response to acetylcholine, nitroprusside and felodipine was also similar. However, the response to caffeine was increased in vessels from the hypertensive patients, although the relationship between the dependence on the effect of calcium on the behaviour of arteries from hypertensives and controls was similar in the presence of prazosin and yohimbine. CONCLUSIONS: The altered morphology of subcutaneous resistance arteries from hypertensives is not caused by a change in the elastic characteristics of the wall material. The data support our previous observation of abnormal calcium handling in vessels from hypertensives, although they do not support the hypothesis that a generalized abnormality in endothelium-dependent or endothelium-independent relaxation is of importance in essential hypertension.
Assuntos
Hipertensão/fisiopatologia , Pele/irrigação sanguínea , Acetilcolina/farmacologia , Adulto , Artérias/efeitos dos fármacos , Artérias/fisiopatologia , Cafeína/farmacologia , Elasticidade , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Nitroprussiato/farmacologia , Vasoconstrição/efeitos dos fármacosRESUMO
The morphological and functional characteristics of isolated subcutaneous resistance vessels (about 230 microns internal diameter) from 13 patients treated for essential hypertension for a median period of 14 months and from 15 matched normotensive controls were examined. The blood pressure of the patients and the controls were not significantly different at the time of examination. However, although compared with the controls, the lumen diameter of the vessels from the patients was not significantly different, the media thickness to lumen diameter ratio was 19% greater. Furthermore, although there was no difference in the active pressure response of the vessels from the two groups, the vessels from the patients had a lower sensitivity to calcium, relaxed faster after a contraction and the sensitivity to exogenous noradrenaline shifted more to the left with cocaine. Since the abnormalities found here have previously also been found in vessels from patients with untreated essential hypertension, the study suggests that despite antihypertensive treatment to normotensive levels for about 1 year, some morphological as well as functional characteristics of the resistance arteries are not fully normalized. This could have consequences for the prognosis of essential hypertension.
Assuntos
Anti-Hipertensivos/uso terapêutico , Hipertensão/patologia , Músculo Liso Vascular/patologia , Resistência Vascular , Artérias/patologia , Feminino , Humanos , Hipertensão/tratamento farmacológico , Masculino , Pessoa de Meia-Idade , Pele/irrigação sanguínea , Fatores de TempoRESUMO
A total of 25 patients with newly diagnosed or poorly controlled essential hypertension were randomly selected from a larger group referred to hospital. Treatment was initiated with perindopril (4-8 mg orally). If normotension was not achieved, isradipine (5-10 mg orally) was added and, if necessary, hydralazine was added. Before treatment and at the end of a 9-month period of normotension (diastolic blood pressure < or = 90 mm Hg), 24-hour blood pressure and echocardiographic measurements were performed and resistance artery structure was determined. A total of 20 age- and sex-matched normotensives were used as controls. During antihypertensive treatment, mean blood pressure was reduced from 128 +/- 11 to 103 +/- 6 mm Hg. Left ventricular mass was reduced from 300 +/- 76 to 198 +/- 54 g. The media:lumen ratio of the resistance arteries decreased from 9.8 +/- 2.6% to 7.8 +/- 1.9%; control subjects exhibited a media:lumen ratio of the same magnitude (7.9 +/- 2.0%). Results indicate that a perindopril-based regimen is extremely efficient in normalizing resistance artery and cardiac ventricular structures within one year of treatment. The impact of these findings on the excess cardiovascular morbidity and mortality in arterial hypertension still remains to be demonstrated.