RESUMO
AIM: Exercise and insulin each increase microvascular blood flow and enhance glucose disposal in skeletal muscle. We have reported that insulin-mediated microvascular recruitment in a diet-induced model of insulin resistance (high-fat feeding for 4 weeks) is markedly impaired; however, the effect of muscle contraction in this model has not been previously explored. METHODS: We fed rats either normal (ND, 10% calories from fat) or high-fat (HFD, 60% calories from fat) diets ad libitum for 4-8 weeks. Animals were then anaesthetized and one hindlimb electrically stimulated to contract at 0.05, 0.1 and 2 Hz (field stimulation, 30 V, 0.1 ms duration) in 15 min stepwise increments. Femoral artery blood flow (Transonic flow probe), muscle microvascular blood flow (hindleg metabolism of 1-methylxanthine and contrast-enhanced ultrasound) and muscle glucose disposal (uptake of radiolabelled 2-deoxy-d-glucose and hindleg glucose disappearance) were measured. RESULTS: Both ND and HFD rats received the same voltage across the leg and consequently developed the same muscle tension. Femoral artery blood flow in the contracting leg increased during 2 Hz contraction, but not during the lower frequencies and these effects were similar between ND and HFD rats. Muscle microvascular blood flow significantly increased in a contraction frequency-dependent manner, and preceded increases in total limb blood flow and these effects were similar between ND and HFD rats. Muscle glucose disposal was markedly elevated during 2 Hz contraction and was comparable between ND and HFD rats. CONCLUSION: Contraction-mediated muscle microvascular recruitment and glucose uptake are not impaired in the HFD insulin resistant rat.
Assuntos
Estimulantes do Sistema Nervoso Central/farmacologia , Artéria Femoral/fisiopatologia , Membro Posterior/irrigação sanguínea , Resistência à Insulina , Contração Muscular , Xantinas/farmacologia , Animais , Glicemia/metabolismo , Dieta Hiperlipídica , Estimulação Elétrica , Masculino , Músculo Esquelético , Ratos , Ratos Sprague-Dawley , Ratos Wistar , Fluxo Sanguíneo Regional/efeitos dos fármacosRESUMO
There is considerable support for the concept that insulin-mediated increases in microvascular blood flow to muscle impact significantly on muscle glucose uptake. Since the microvascular blood flow increases with insulin have been shown to be nitric oxide-dependent inhibition of cGMP-degrading phosphodiesterases (cGMP PDEs) is predicted to enhance insulin-mediated increases in microvascular perfusion and muscle glucose uptake. Therefore, we studied the effects of the pan-cGMP PDE inhibitor zaprinast on the metabolic and vascular actions of insulin in muscle. Hyperinsulinemic euglycemic clamps (3 mU·min(-1)·kg(-1)) were performed in anesthetized rats and changes in microvascular blood flow assessed from rates of 1-methylxanthine metabolism across the muscle bed by capillary xanthine oxidase in response to insulin and zaprinast. We also characterized cGMP PDE isoform expression in muscle by real-time PCR and immunostaining of frozen muscle sections. Zaprinast enhanced insulin-mediated microvascular perfusion by 29% and muscle glucose uptake by 89%, while whole body glucose infusion rate during insulin infusion was increased by 33% at 2 h. PDE2, -9, and -10 were the major isoforms expressed at the mRNA level in muscle, while PDE1B, -9A, -10A, and -11A proteins were expressed in blood vessels. Acute administration of the cGMP PDE inhibitor zaprinast enhances muscle microvascular blood flow and glucose uptake response to insulin. The expression of a number of cGMP PDE isoforms in skeletal muscle suggests that targeting specific cGMP PDE isoforms may provide a promising avenue for development of a novel class of therapeutics for enhancing muscle insulin sensitivity.
Assuntos
Nucleotídeo Cíclico Fosfodiesterase do Tipo 2/antagonistas & inibidores , Insulina/metabolismo , Músculo Esquelético , Purinonas/farmacologia , Animais , Aorta/citologia , Glicemia/metabolismo , Células Cultivadas , Nucleotídeo Cíclico Fosfodiesterase do Tipo 2/metabolismo , Metabolismo Energético/efeitos dos fármacos , Metabolismo Energético/fisiologia , Técnica Clamp de Glucose , Hiperinsulinismo/metabolismo , Masculino , Microcirculação/efeitos dos fármacos , Microcirculação/fisiologia , Músculo Esquelético/irrigação sanguínea , Músculo Esquelético/efeitos dos fármacos , Músculo Esquelético/metabolismo , Músculo Liso Vascular/citologia , Inibidores de Fosfodiesterase/farmacologia , Ratos , Ratos WistarRESUMO
AIM: Intracerebroventricular (ICV) administration of a nitric oxide synthase (NOS) inhibitor to rats has been reported to raise blood pressure (BP) and cause insulin resistance, suggestive of a central effect of insulin that is NO dependent. Herein we test whether ICV insulin has peripheral haemodynamic and metabolic effects and whether peripheral effects of systemic insulin are affected by the ICV administration of the NOS inhibitor N(G) -methyl-l-arginine (l-NMMA). METHODS: Anaesthetized rats were fitted with an ICV cannula for insulin, artificial cerebrospinal fluid (aCSF) or l-NMMA infusion. Rats receiving ICV l-NMMA (500 µg) underwent systemic insulin clamp (10 mU/min/kg) or saline treatment for 70 min and were compared with animals receiving an equal amount of l-NMMA infused systemically. RESULTS: ICV aCSF or insulin (135 mU/min/kg brain) for 70 min or systemic l-NMMA (500 µg) had no effect on BP, heart rate (HR), femoral blood flow (FBF), glucose infusion rate, muscle 2-deoxyglucose uptake, microvascular perfusion or plasma insulin. However, ICV l-NMMA reduced systemic insulin-mediated increases in FBF (2.05 ± 0.08 to 1.55 ± 0.15 ml/min), 2-deoxyglucose uptake (17.7 ± 0.15 to 10.0 ± 0.03 µg/g/min) and microvascular perfusion (10.5 ± 0.5 to 6.6 ± 1.1 mol/min) (each mean ± SE, p < 0.05); plasma insulin, HR and BP were unaffected. CONCLUSIONS: Central insulin administration had no effect on skeletal muscle haemodynamics or glucose metabolism. However, systemic insulin-mediated increases in limb blood flow, muscle microvascular perfusion and glucose uptake may be regulated by a central pathway that is NO dependent.
Assuntos
Glicemia/metabolismo , Hipoglicemiantes/administração & dosagem , Insulina/administração & dosagem , Músculo Esquelético/irrigação sanguínea , Músculo Esquelético/efeitos dos fármacos , ômega-N-Metilarginina/administração & dosagem , Animais , Hemodinâmica , Hipoglicemiantes/farmacologia , Injeções Intraventriculares , Insulina/farmacologia , Masculino , Perfusão , Ratos , Ratos Wistar , ômega-N-Metilarginina/farmacologiaRESUMO
AIM: The aetiology of the development of type 2 diabetes remains unresolved. In the present study, we assessed whether an impairment of insulin-mediated microvascular perfusion occurs early in the onset of insulin resistance. MATERIALS AND METHODS: Hooded Wistar rats were fed either a normal diet (ND) or a high-fat diet (HFD) for 4 weeks. Anaesthetized animals were subjected to an isoglycaemic hyperinsulinaemic clamp (3 or 10 mU/min/kg x 2 h), and measurements were made of glucose infusion rate (GIR), hindleg glucose uptake, muscle glucose uptake by 2-deoxy-d-glucose (R'g), glucose appearance (Ra), glucose disappearance (Rd), femoral blood flow (FBF) and hindleg 1-methylxanthine disappearance (1-MXD, an index of microvascular perfusion). RESULTS: Compared with ND-fed animal, HFD feeding led to a mild increase in fasting plasma glucose and plasma insulin, without an increase in total body weight. During the clamps, HFD rats showed an impairment of insulin-mediated action on GIR, hindleg glucose uptake, R'g, Ra, Rd and FBF, with a greater loss of insulin responsiveness at 3 mU/min/kg than at 10 mU/min/kg. The HFD also impaired insulin-mediated microvascular perfusion as assessed by 1-MXD. Interestingly, 1-MXD was the only measurement that remained unresponsive to the higher dose of 10 mU/min/kg insulin. CONCLUSIONS: We conclude that the early stage of insulin resistance is characterized by an impairment of the insulin-mediated microvascular responses in skeletal muscle. This is likely to cause greater whole body insulin resistance by limiting the delivery of hormones and nutrients to muscle.
Assuntos
Gorduras na Dieta/administração & dosagem , Hipoglicemiantes/farmacologia , Resistência à Insulina/fisiologia , Insulina/farmacologia , Microcirculação/fisiologia , Músculo Esquelético/irrigação sanguínea , Animais , Glicemia/metabolismo , Masculino , Músculo Esquelético/metabolismo , Músculo Liso Vascular/fisiologia , Ratos , Ratos Wistar , Fluxo Sanguíneo Regional/fisiologiaRESUMO
The uptake of 2-deoxyglucose by perfused rat hearts was compared to the distribution of the insulin-regulatable glucose transporter (GLUT4) in membrane preparations from the same hearts. The hearts were treated with the alpha-adrenergic combination of epinephrine + propranolol, the beta-adrenergic agonist isoproterenol, high (8 mM) Ca2+ concentrations, insulin and the alpha adrenergic combination or insulin alone. Epinephrine (1 microM) + propranolol (10 microM), isoproterenol (10 microM), high Ca2+, insulin (1 microM) + epinephrine (1 microM) + propranolol (10 microM) and insulin (1 microM) each led to an increase in 2-deoxyglucose uptake and a shift in the recovery of the GLUT4 from a high-speed pellet membrane fraction (putatively intracellular) to a low-speed pellet membrane fraction (putatively sarcolemmal). There were significant correlations (r = -0.673, P less than 0.001) between the stimulation of 2-deoxyglucose uptake and the loss of GLUT4 from the intracellular membrane fraction, or the increase in the sarcolemmal fraction. The data provide evidence that the GLUT4 is translocated by agents that stimulate glucose transport in heart, and therefore this mechanism is not restricted to insulin.
Assuntos
Cálcio/farmacologia , Catecolaminas/farmacologia , Glucose/metabolismo , Proteínas de Transporte de Monossacarídeos/metabolismo , Miocárdio/metabolismo , 5'-Nucleotidase/metabolismo , Animais , Transporte Biológico/efeitos dos fármacos , Galactosiltransferases/metabolismo , Técnicas In Vitro , Insulina/farmacologia , Masculino , Ratos , Sarcolema/metabolismo , Frações Subcelulares/químicaRESUMO
Insulin (0.1 microM) and 1 microM epinephrine each increased the uptake and phosphorylation of 2-deoxyglucose by the perfused rat heart by increasing the apparent Vmax without altering the Km. Isoproterenol (10 microM), 50 microM methoxamine and 10 mM CaCl2 also increased uptake. Lowering of the perfusate Ca2+ concentration from 1.27 to 0.1 mM Ca2+, addition of the Ca2+ channel blocker nifedipine (1 microM) or addition of 1.7 mM EGTA decreased the basal rate of uptake of 2-deoxyglucose and prevented the stimulation due to 1 microM epinephrine. Stimulation of 2-deoxyglucose uptake by 0.1 microM insulin was only partly inhibited by Ca2+ omission, nifedipine or 1 mM EGTA. Half-maximal stimulation of 2-deoxyglucose uptake by insulin occurred at 2 nM and 0.4 nM for medium containing 1.27 and 0.1 mM Ca2+, respectively. Maximal concentrations of insulin (0.1 microM) and epinephrine (1 microM) were additive for glucose uptake and lactate output but were not additive for uptake of 2-deoxyglucose. Half-maximal stimulation of 2-deoxyglucose uptake by epinephrine occurred at 0.2 microM but maximal concentrations of epinephrine (e.g., 1 microM) gave lower rates of 2-deoxyglucose uptake than that attained by maximal concentrations of insulin. The addition of insulin increased uptake of 2-deoxyglucose at all concentrations of epinephrine but epinephrine only increased uptake at sub-maximal concentrations of insulin. The role of Ca2+ in signal reversal was also studied. Removal of 1 microM epinephrine after a 10 min exposure period resulted in a rapid return of contractility to basal values but the rate of 2-deoxyglucose uptake increased further and remained elevated at 20 min unless the Ca2+ concentration was lowered to 0.1 mM or nifedipine (1 microM) was added. Similarly, removal of 0.1 microM insulin after a 10 min exposure period did not affect the rate of 2-deoxyglucose uptake, which did not return to basal values within 20 min unless the concentration of Ca2+ was decreased to 0.1 mM. Insulin-mediated increase in 2-deoxyglucose uptake at 0.1 mM Ca2+ reversed upon hormone removal. It is concluded that catecholamines mediate a Ca2+-dependent increase in 2-deoxyglucose transport from either alpha or beta receptors. Insulin has both a Ca2+-dependent and a Ca2+-independent component. Reversal studies suggest an additional role for Ca2+ in maintaining the activated transport state when activated by either epinephrine or insulin.
Assuntos
Cálcio/farmacologia , Desoxiaçúcares/metabolismo , Desoxiglucose/metabolismo , Epinefrina/farmacologia , Insulina/farmacologia , Miocárdio/metabolismo , Animais , Relação Dose-Resposta a Droga , Coração/efeitos dos fármacos , Isoproterenol/farmacologia , Cinética , Masculino , Metoxamina/farmacologia , Fosforilação , Ratos , Fatores de TempoRESUMO
The vascular actions of insulin may contribute to the increase in glucose uptake by skeletal muscle. We have recently shown that when capillary recruitment by insulin is blocked in vivo, an acute state of insulin resistance is induced. Another agent that may have vascular effects is the inflammatory cytokine tumor necrosis factor-alpha (TNF-alpha), which has been reported to play an important role in the insulin resistance of obesity, type 2 diabetes, and sepsis in both animals and humans. Thus, in the present study, we have investigated the effect of an intravenous 3-h TNF treatment (0.5 microg x h(1) x kg(-1)) in control and euglycemic-hyperinsulinemic-clamped (10 mU x min(-1) x kg(-1) for 2 h) anesthetized rats. Hind-leg glucose uptake, muscle uptake of 2-deoxyglucose (2-DG), femoral blood flow (FBF), vascular resistance (VR), and capillary recruitment as measured by metabolism of infused 1-methylxanthine (1-MX) were assessed. Insulin alone caused a significant (P < 0.05) increase in FBF (1.7-fold) and capillary recruitment (2.5-fold), with a significant decrease in VR. In addition, hind-leg glucose uptake was increased (fourfold), as was 2-DG uptake in the soleus and plantaris muscles. TNF completely prevented the insulin-mediated changes in FBF, VR, and capillary recruitment and significantly reduced (P < 0.05) the insulin-mediated increase in total hind-leg glucose uptake (by 61%) and muscle 2-DG uptake (by at least 50%). TNF alone had no significant effect on any of these variables. It is concluded that acute administration in vivo of TNF completely blocks the hemodynamic actions of insulin on rat skeletal muscle vasculature and blocks approximately half of the glucose uptake by muscle. It remains to be determined whether these two effects are interdependent.
Assuntos
Capilares/efeitos dos fármacos , Glucose/metabolismo , Insulina/farmacologia , Músculo Esquelético/irrigação sanguínea , Músculo Esquelético/metabolismo , Fator de Necrose Tumoral alfa/farmacologia , Animais , Velocidade do Fluxo Sanguíneo , Glicemia/análise , Pressão Sanguínea , Capilares/fisiologia , Desoxiglucose/metabolismo , Artéria Femoral/fisiologia , Veia Femoral/fisiologia , Técnica Clamp de Glucose , Insulina/sangue , Resistência à Insulina , Masculino , Músculo Esquelético/efeitos dos fármacos , Ratos , Ratos Wistar , Fator de Necrose Tumoral alfa/administração & dosagem , Resistência Vascular/efeitos dos fármacos , Xantinas/administração & dosagem , Xantinas/metabolismoRESUMO
Insulin-induced increases in blood flow are hypothesized to enhance overall glucose uptake by skeletal muscle. Whether the insulin-mediated changes in blood flow are associated with altered blood flow distribution and increased capillary recruitment in skeletal muscle is not known. In the present study, the effects of insulin on hemodynamic parameters in rat skeletal muscle in vivo were investigated. Mean arterial blood pressure, heart rate, femoral blood flow, hind leg vascular resistance, and glucose uptake were measured in control and euglycemic insulin-clamped (10 mU x min(-1) x kg[-1]) anesthetized rats. Blood flow distribution within the hind leg muscles was assessed by measuring the metabolism of 1-methylxanthine (1-MX), an exogenously added substrate for capillary xanthine oxidase. Insulin treatment had no effect on heart rate but significantly increased arterial blood pressure (12 mmHg) and femoral blood flow (80%) and decreased hind leg vascular resistance (31%). Changes were similar in magnitude and in time of onset to those reported in humans. Insulin treatment increased hind leg glucose uptake approximately fourfold and also increased hind leg 1-MX metabolism by 50%, suggesting increased exposure to endothelial xanthine oxidase. To ascertain whether the increased 1-MX metabolism was simply due to increased bulk femoral blood flow, epinephrine was infused at a dose (0.125 microg x min(-) x kg[-1]) chosen to match the insulin-induced increase in femoral blood flow. This dose of epinephrine had no significant effects on arterial blood pressure or heart rate but increased femoral blood flow and lowered hind leg vascular resistance to a similar extent as insulin. Epinephrine did not significantly alter 1-MX metabolism as compared with control animals. These results demonstrate that insulin increases total hind leg blood flow and metabolism of 1-MX, suggesting a recruitment of capillary blood flow in rat hind leg not mimicked by epinephrine.
Assuntos
Hemodinâmica/efeitos dos fármacos , Insulina/farmacologia , Músculo Esquelético/irrigação sanguínea , Animais , Epinefrina/farmacologia , Glucose/metabolismo , Membro Posterior , Ratos , Ratos Sprague-Dawley , Fluxo Sanguíneo Regional/efeitos dos fármacos , Resistência Vascular/efeitos dos fármacos , Xantinas/metabolismoRESUMO
Insulin-mediated changes in blood flow are associated with altered blood flow distribution and increased capillary recruitment in skeletal muscle. Studies in perfused rat hindlimb have shown that muscle metabolism can be regulated by vasoactive agents that control blood flow distribution within the hindlimb. In the present study, the effects of a vasoconstrictive agent that has no direct effect on skeletal muscle metabolism but that alters perfusion distribution in rat hindlimb was investigated in vivo to determine its effects on insulin-mediated vascular action and glucose uptake. We measured the effects of alpha-methylserotonin (alpha-met5HT) on mean arterial blood pressure, heart rate, femoral blood flow, hindlimb vascular resistance, and glucose uptake in control and euglycemic insulin-clamped (10 mU x min(-1) x kg(-1)) anesthetized rats. Blood flow distribution within the hindlimb muscles was assessed by measuring the metabolism of 1-methylxanthine (1-MX), an exogenously added substrate for capillary xanthine oxidase. Alpha-met5HT (20 microg x min(-1) x kg(-1)) infusion alone increased mean arterial blood pressure by 25% and increased hindlimb vascular resistance but caused no change in femoral blood flow. These changes were associated with decreased hindlimb 1-MX metabolism indicating less capillary flow. Insulin infusion caused decreased hindlimb vascular resistance that was associated with increased femoral blood flow and 1-MX metabolism. Treatment with alpha-met5HT infusion commenced before insulin infusion prevented the increase in femoral blood flow and inhibited the stimulation of 1-MX metabolism. Alpha-met5HT infusion had no effect on hindlimb glucose uptake but markedly inhibited the insulin stimulation of glucose uptake (P < 0.05) and was associated with decreased glucose infusion rates to maintain euglycemia (P < 0.05). A significant correlation (P < 0.05) was observed between 1-MX metabolism and hindlimb glucose uptake but not between femoral blood flow and glucose uptake. The results indicate that in vivo, certain types of vasoconstriction in muscle such as elicited by 5HT2 agonists, which prevent normal insulin recruitment of capillary flow, cause impaired muscle glucose uptake. Moreover, if vasoconstriction of this kind results from stress-induced increase in sympathetic outflow, then this may provide a clue as to the link between hypertension and insulin resistance that is often observed in humans.
Assuntos
Hemodinâmica/fisiologia , Resistência à Insulina/fisiologia , Insulina/farmacologia , Serotonina/análogos & derivados , Vasoconstritores/farmacologia , Alopurinol/farmacologia , Animais , Glicemia/metabolismo , Pressão Sanguínea/efeitos dos fármacos , Pressão Sanguínea/fisiologia , Técnica Clamp de Glucose , Hemodinâmica/efeitos dos fármacos , Membro Posterior/irrigação sanguínea , Humanos , Infusões Intravenosas , Insulina/administração & dosagem , Masculino , Ratos , Ratos Sprague-Dawley , Fluxo Sanguíneo Regional/efeitos dos fármacos , Fluxo Sanguíneo Regional/fisiologia , Serotonina/farmacologia , Agonistas do Receptor de Serotonina/farmacologia , Resistência Vascular/efeitos dos fármacos , Resistência Vascular/fisiologiaRESUMO
Exercise training is considered to be beneficial in the treatment and prevention of insulin insensitivity, and much of the effect occurs in muscle. We have recently shown that capillary recruitment by insulin in vivo is associated with and may facilitate insulin action to increase muscle glucose uptake. In the present study, we examined the effect of 14 days of voluntary exercise training on euglycemic-hyperinsulinemic clamped (10 mU. min(-1). kg(-1) for 2 h), anesthetized rats. Whole-body glucose infusion rate (GIR), hindleg glucose uptake, femoral blood flow (FBF), vascular resistance, and capillary recruitment, as measured by metabolism of infused 1-methylxanthine (1-MX), were assessed. In sedentary animals, insulin caused a significant (P < 0.05) increase in FBF (1.6-fold) and capillary recruitment (1.7-fold) but a significant decrease in vascular resistance. In addition, hindleg glucose uptake was increased (4.3-fold). Exercise training increased insulin-mediated GIR (24%), hindleg glucose uptake (93%), and capillary recruitment (62%) relative to sedentary animals. Neither capillary density nor total xanthine-oxidase activity in skeletal muscle were increased as a result of the training regimen used. We concluded that exercise training improves insulin-mediated increases in capillary recruitment in combination with augmented muscle glucose uptake. Increased insulin-mediated glucose uptake may in part result from the improved hemodynamic control attributable to exercise training.
Assuntos
Capilares/fisiologia , Glucose/metabolismo , Insulina/farmacologia , Músculo Esquelético/irrigação sanguínea , Músculo Esquelético/efeitos dos fármacos , Esforço Físico , Animais , Velocidade do Fluxo Sanguíneo , Glicemia/metabolismo , Fêmur/fisiologia , Glucose/administração & dosagem , Técnica Clamp de Glucose , Membro Posterior , Insulina/sangue , Masculino , Músculo Esquelético/metabolismo , Tamanho do Órgão , Ratos , Ratos Wistar , Resistência Vascular , Xantina Oxidase/metabolismo , Xantinas/metabolismoRESUMO
Despite intensive study, the relation between insulin's action on blood flow and glucose metabolism remains unclear. Insulin-induced changes in microvascular perfusion, independent from effects on total blood flow, could be an important variable contributing to insulin's metabolic action. We hypothesized that modest, physiologic increments in plasma insulin concentration alter microvascular perfusion in human skeletal muscle and that these changes can be assessed using contrast-enhanced ultrasound (CEU), a validated method for quantifying flow by measurement of microvascular blood volume (MBV) and microvascular flow velocity (MFV). In the first protocol, 10 healthy, fasting adults received insulin (0.05 mU. kg(-1). min(-1)) via a brachial artery for 4 h under euglycemic conditions. At baseline and after insulin infusion, MBV and MFV were measured by CEU during continuous intravenous infusion of albumin microbubbles with intermittent harmonic ultrasound imaging of the forearm deep flexor muscles. In the second protocol, 17 healthy, fasting adults received a 4-h infusion of either insulin (0.1 mU. kg(-1). min(-1), n = 9) or saline (n = 8) via a brachial artery. Microvascular volume was assessed in these subjects by an alternate CEU technique using an intra-arterial bolus injection of albumin microbubbles at baseline and after the 4-h infusion. With both protocols, muscle glucose uptake, plasma insulin concentration, and total blood flow to the forearm were measured at each stage. In protocol 2 subjects, tissue extraction of 1-methylxanthine (1-MX) was measured as an index of perfused capillary volume. Caffeine, which produces 1-MX as a metabolite, was administered to these subjects before the study to raise plasma 1-MX levels. In protocol 1 subjects, insulin increased muscle glucose uptake (180%, P < 0.05) and MBV (54%, P < 0.01) and decreased MFV (-42%, P = 0.07) in the absence of significant changes in total forearm blood flow. In protocol 2 subjects, insulin increased glucose uptake (220%, P < 0.01) and microvascular volume (45%, P < 0.05) with an associated moderate increase in total forearm blood flow (P < 0.05). Using forearm 1-MX extraction, we observed a trend, though not significant, toward increasing capillary volume in the insulin-treated subjects. In conclusion, modest physiologic increments in plasma insulin concentration increased microvascular blood volume, indicating altered microvascular perfusion consistent with a mechanism of capillary recruitment. The increases in microvascular (capillary) volume (despite unchanged total blood flow) indicate that the relation between insulin's vascular and metabolic actions cannot be fully understood using measurements of bulk blood flow alone.
Assuntos
Capilares/fisiologia , Insulina/sangue , Músculo Esquelético/irrigação sanguínea , Adulto , Albuminas/administração & dosagem , Velocidade do Fluxo Sanguíneo , Glicemia/metabolismo , Volume Sanguíneo , Artéria Braquial , Cafeína/sangue , Feminino , Glucose/metabolismo , Humanos , Insulina/administração & dosagem , Ácido Láctico/metabolismo , Masculino , Músculo Esquelético/metabolismo , Consumo de Oxigênio , Teofilina/sangue , Xantinas/sangue , Xantinas/metabolismoRESUMO
Supraphysiological doses of insulin enhance total limb blood flow and recruit capillaries in skeletal muscle. Whether these processes change in response to physiological hyperinsulinemia is uncertain. To examine this, we infused either saline (n = 6) or insulin (euglycemic clamp, 3.0 mU x min(-1) x kg(-1), n = 9) into anesthetized rats for 120 min. Femoral artery flow was monitored continuously using a Doppler flow probe, and muscle microvascular recruitment was assessed by metabolism of infused 1-methylxanthine (1-MX) and by contrast-enhanced ultrasound (CEU). Insulin infusion raised plasma insulin concentrations by approximately 10-fold. Compared with saline, physiological hyperinsulinemia increased femoral artery flow (1.02 +/- 0.10 vs. 0.68 +/- 0.09 ml/min; P < 0.05), microvascular recruitment (measured by 1-MX metabolism [6.6 +/- 0.5 vs. 4.5 +/- 0.48 nmol/min; P < 0.05] as well as by CEU [167.0 +/- 39.8 vs. 28.2 +/- 13.8%; P < 0.01]), and microvascular flow velocity (beta, 0.14 +/- 0.02 vs. 0.09 +/- 0.02 s(-1)). Subsequently, we studied the time dependency of insulin's vascular action in a second group (n = 5) of animals. Using CEU, microvascular volume was measured at 0, 30, and 90 min of insulin infusion. Insulin augmented microvascular perfusion within 30 min (52.8 +/- 14.8%), and this persisted at 90 min (64.6 +/- 9.9%). Microvascular recruitment occurred without changes to femoral artery flow or beta. We conclude that insulin increases tissue perfusion by recruiting microvascular beds, and at physiological concentrations this precedes increases in total muscle blood flow by 60-90 min.
Assuntos
Capilares/fisiologia , Hiperinsulinismo/fisiopatologia , Microcirculação/fisiologia , Músculo Esquelético/irrigação sanguínea , Ácido Úrico/análogos & derivados , Animais , Biotransformação , Capilares/fisiopatologia , Membro Posterior , Infusões Intravenosas , Insulina/administração & dosagem , Insulina/metabolismo , Insulina/farmacologia , Cinética , Masculino , Ratos , Ratos Sprague-Dawley , Fluxo Sanguíneo Regional/efeitos dos fármacos , Ácido Úrico/farmacocinética , Xantina Oxidase/metabolismo , Xantinas/farmacocinéticaRESUMO
Electrical stimulation of the sciatic nerve of the anaesthetized rat in vivo led to a time-dependent translocation of protein kinase C from the muscle cytosol to the particulate fraction. Maximum activity of protein kinase C in the particulate fraction occurred after 2 min of intermittent short tetanic contractions of the gastrocnemius-plantaris-soleus muscle group and coincided with the loss of activity from the cytosol. Translocation of protein kinase C may imply a role for this kinase in contraction-initiated changes in muscle metabolism.
Assuntos
Músculos/enzimologia , Proteína Quinase C/metabolismo , Animais , Transporte Biológico , Cálcio/farmacologia , Citosol/enzimologia , Cinética , Masculino , Contração Muscular , Fosfolipídeos/farmacologia , Ratos , Ratos Endogâmicos , Frações Subcelulares/enzimologiaRESUMO
Recent findings indicate that glucose uptake by contracting hindlimb #Acta Physiol. Scand. (1982) 116, 215-222 #and heart #Biochem. Biophys. Res. Commun. (1982) 108, 124-131 # of the rat is stimulated by epinephrine acting through alpha-adrenergic mechanisms. Since in exercise hepatic glucose output may be increased markedly by activation of alpha-adrenergic receptors and matched by the increase in muscle glucose uptake (maintaining blood glucose levels relatively constant), it is now proposed that a general coordination of glucose metabolism may operate via alpha-adrenergic receptor mechanisms. The basis for this proposal is discussed.
Assuntos
Catecolaminas/farmacologia , Glucose/metabolismo , Glicólise/efeitos dos fármacos , Glicogênio Hepático/metabolismo , Músculos/metabolismo , Esforço Físico , Receptores Adrenérgicos alfa/fisiologia , Receptores Adrenérgicos/fisiologia , Animais , Gluconeogênese , Miocárdio/metabolismo , RatosRESUMO
Vasoconstriction mediated by serotonin (5-HT) inhibits muscle metabolism in resting constant-flow-perfused rat hindlimb and may do so by vascular shunting. In the present study, the effects of 5-HT on tension development and contraction-induced oxygen uptake by the sciatic nerve-stimulated gastrocnemius-plantaris-soleus muscle group of the perfused rat hindlimb and tension development by electrically stimulated isolated incubated soleus and extensor digitorum longus muscles were examined. In both erythrocyte and erythrocyte-free perfusions, 0.25 microM 5-HT increased perfusion pressure and markedly decreased contraction-induced tension, oxygen uptake, and lactate release. The release of metabolic vasodilators from exercising skeletal muscle did not appear to affect 5-HT-mediated vasoconstriction; rather, vascular resistance increased during the period of muscle contraction. In contrast, vasoconstriction during muscle contraction mediated by alpha-adrenoceptor stimulation did not impair tension and was partially overcome by metabolic vasodilators. In addition, contraction of isolated incubated soleus and extensor digitorum longus muscles was not affected by 5-HT addition to the incubation medium. We conclude that 5-HT impairs contractility of working muscle during the aerobic phase by limiting oxygen delivery through redistributing perfusate flow. The results are consistent with a vasoconstrictor action of 5-HT on larger vessels, perhaps at feed arteries external to the working muscle. When constricted by 5-HT, these vessels are apparently insensitive to metabolic vasodilatation.
Assuntos
Músculo Esquelético/fisiologia , Serotonina/fisiologia , Vasoconstrição/fisiologia , Aerobiose/fisiologia , Animais , Bovinos , Estimulação Elétrica , Membro Posterior/efeitos dos fármacos , Membro Posterior/fisiologia , Técnicas In Vitro , Lactatos/metabolismo , Ácido Láctico , Masculino , Contração Muscular/efeitos dos fármacos , Contração Muscular/fisiologia , Consumo de Oxigênio/fisiologia , Ratos , Ratos Wistar , Serotonina/farmacologia , Vasoconstrição/efeitos dos fármacosRESUMO
The effects of different vasomodulators on lactate release by the constant-flow-perfused rat hindlimb were examined and compared with that by perfused mesenteric artery, incubated preparations of aortas, soleus and epitrochlearis muscles, and perifused soleus muscles. Infusion of vasopressin (0.5 nM), angiotensin II (5 nM), norepinephrine (50 nM), and methoxamine (10 microM) into the hindlimbs of 180- to 200-g rats increased the perfusion pressure by 112-167% from 30.4 +/- 0.8 mmHg, O2 consumption by 26-68% from 6.4 +/- 0.2 mumol.g-1 x h-1, and lactate efflux by 148-380% from 5.41 +/- 0.25 mumol.g-1 x h-1. Hindlimbs of 100- to 120-g rats responded similarly to angiotensin II. Isoproterenol (1 microM) had no effect on O2 uptake or perfusion pressure but increased lactate release by 118%. Nitroprusside (0.5 mM) markedly inhibited the vasoconstrictor-mediated increases in lactate release, perfusion pressure, and O2 consumption by the hindlimb but had no effect on isoproterenol-mediated lactate efflux. Serotonin (6.7 microM) increased lactate release from the perfused mesenteric artery by 120% from 5.48 mol.g-1 x h-1. Lactate release by incubated aorta was increased by angiotensin II (50 nM), isoproterenol (1 microM), and mechanical stretch. The increase mediated by angiotensin II was blocked by glycerol trinitrate (2.2 microM), which had no effect on lactate release by isoproterenol. Neither angiotensin II (5 nM) nor vasopressin (0.5 nM) increased lactate release from incubated soleus and epitrochlearis muscles; however, lactate release was increased by isoproterenol, and this increase was unaffected by glycerol trinitrate (2.2 microM).(ABSTRACT TRUNCATED AT 250 WORDS)
Assuntos
Lactatos/metabolismo , Músculos/metabolismo , Vasoconstritores/farmacologia , Animais , Membro Posterior , Técnicas In Vitro , Masculino , Músculo Liso/efeitos dos fármacos , Músculo Liso/metabolismo , Músculo Liso Vascular/efeitos dos fármacos , Músculo Liso Vascular/metabolismo , Músculos/efeitos dos fármacos , Nitroprussiato/farmacologia , Consumo de Oxigênio/efeitos dos fármacos , Perfusão , Ratos , Ratos Wistar , Serotonina/fisiologiaRESUMO
Systemic hypertension of mild to moderate degree is often associated with obesity. The hypothesis is that over-eating leads to increased sympathetic activity targeted at the peripheral vasculature as well as other tissues in an attempt (that in many cases may be futile) to stimulate facultative thermogenesis and burn-off the excess energy. This hypothesis represents an important modification of one proposed by Landsberg and is supported by: 1) recent observations that carbohydrate feeding to humans specifically increases muscle sympathetic vasoconstrictor activity in the peroneal nerve, and 2) studies with animal models in which active vasoconstriction in the limbs and elsewhere is associated with marked increases in oxygen consumption (energy expenditure).
Assuntos
Regulação da Temperatura Corporal/fisiologia , Homeostase/fisiologia , Hipertensão/fisiopatologia , Obesidade/complicações , Animais , Feminino , Humanos , Hipertensão/etiologia , Masculino , Obesidade/fisiopatologia , RatosRESUMO
We have recently shown that the vasoconstrictor serotonin (5-HT) inhibits oxygen uptake in perfused hindlimb possibly due to vascular shunting. Thus in the present study the effect of 5-HT on insulin-mediated glucose uptake was assessed. Rat hindlimbs were perfused at constant flow with medium containing 8.3 mM glucose and a tracer amount of 2-deoxy-D-[1-3]glucose (2DG) with and without 10 microM 5-HT, 15 nM insulin and a combination of the two. 5-HT inhibited insulin-mediated stimulation of glucose uptake by 30.4% when added after insulin and 34.4% when added before insulin. In addition, 5-HT inhibited insulin-mediated 2DG uptake by perfused muscles with inhibition ranging from 32% (soleus) to 80% (extensor digitorum longus). The effects of 5-HT on insulin-mediated glucose uptake were partially reversed by vasodilation with carbachol. In contrast to the results for the hindlimb, 10 microM 5-HT had no significant effect on either basal glucose uptake or the stimulation of glucose uptake mediated by 15 nM insulin by isolated incubated soleus or extensor digitorum longus muscles. It is concluded that 5-HT impairs insulin-mediated glucose uptake in the perfused rat hindlimb that may derive from vascular shunting not apparent when muscles are incubated with 5-HT in vitro. These findings may have implications for the link between hypertension and diabetes.
Assuntos
Membro Posterior/irrigação sanguínea , Membro Posterior/metabolismo , Resistência à Insulina/fisiologia , Músculos/irrigação sanguínea , Músculos/metabolismo , Serotonina/farmacologia , Vasoconstritores/farmacologia , Animais , Relação Dose-Resposta a Droga , Glucose/farmacocinética , Técnicas In Vitro , Insulina/farmacologia , Masculino , Músculos/efeitos dos fármacos , Oxigênio/farmacocinética , Consumo de Oxigênio/efeitos dos fármacos , Perfusão , Ratos , Ratos Wistar , Vasoconstrição/efeitos dos fármacos , Vasoconstrição/fisiologiaRESUMO
Vasoconstriction by norepinephrine, angiotensin II and vasopressin in the constant-flow perfused rat hindlimb is associated with increased oxygen uptake and has given rise to the concept of vascular thermogenesis. In the present study serotonin (5-hydroxytryptamine, 5HT) was found to inhibit oxygen uptake by up to 40% in a dose dependent manner whilst inducing vasoconstriction in this model, whereas norepinephrine increased oxygen consumption by up to 100% during vasoconstriction. This contrasted with the perfused isolated rat mesenteric artery arcade in which serotonin stimulated oxygen uptake by up to 130% in association with vasoconstriction in a dose dependent manner similar to the previously described norepinephrine induced vascular thermogenesis in this arterial preparation. In both perfusion systems, changes in pressure and oxygen uptake mediated by serotonin were completely blocked by ketanserin. These results and evidence from dye washout studies suggest that serotonin-mediated vascular thermogenesis, if it occurs in the constant-flow hindlimb, is masked by vascular shunting.
Assuntos
Vasos Sanguíneos/fisiologia , Regulação da Temperatura Corporal , Serotonina/fisiologia , Animais , Masculino , Norepinefrina/fisiologia , Oxigênio/sangue , Ratos , Ratos Endogâmicos , VasoconstriçãoRESUMO
PURPOSE: To assess the results of implantation of secondary, open-loop, flexible, anterior chamber intraocular lenses (IOLs) and compare the findings with those of other published studies. SETTING: A combined ophthalmology and ear, nose, and throat hospital in Middlesbrough, Cleveland, England. METHODS: This retrospective study comprised 81 patients who had secondary implantation of a flexible, open-loop, anterior chamber IOL by the one surgeon. The incidence of postoperative complications was ascertained, and best corrected preoperative and postoperative visual acuities were compared. RESULTS: Two serious complications occurred: one severe loss of vision that is still under investigation and one retinal detachment that was repaired with good residual visual function. Of all 81 patients, 92.5% were within one Snellen line of their preoperative best corrected visual acuity. These results compare favorably with those of other published studies. CONCLUSION: Using an open-loop, flexible, anterior chamber lens for secondary implantation is still an acceptable way to treat aphakia. The poor reputation of these lenses is undeserved.