Efeito dos exercícios resistidos sobre a função endotelial, parâmetros hemodinâmicos e metabólicos, modulação autonômica, biomarcadores inflamatórios, composição corporal e condicionamento físico de adolescentes obesos

Introdução - O exercício aeróbio é considerado uma ferramenta eficaz na redução de fatores de risco cardiometabólico e na melhora da qualidade de vida em adolescentes obesos. No entanto, os benefícios de um programa exclusivo com exercícios resistidos (ER) ainda não foram estabelecidos nesta população. Objetivo - Avaliar o efeito de um programa exclusivo com ER sobre a função endotelial, parâmetros hemodinâmicos e metabólicos, modulação autonômica, biomarcadores inflamatórios, composição corporal e condicionamento físico de adolescentes obesos. Materiais e Métodos - Quarenta adolescentes participaram do estudo e foram divididos em dois grupos de acordo com o nível de adiposidade: grupo controle (C, n = 20; 13 meninos e 7 meninas) e grupo obeso (Ob; n=24; 7 meninos e 17 meninas). O grupo Ob foi submetido a um programa de ER, três vezes por semana, durante três meses. A função endotelial, o perfil metabólico e hemodinâmico, a modulação autonômica, os biomarcadores inflamatórios, a composição corporal e o condicionamento físico foram avaliados antes e ao final da intervenção. Na investigação de nossa hipótese utilizamos: laser-Doppler fluxometria, análises bioquímicas e de células endoteliais circulantes, monitorização da pressão arterial ambulatorial, variabilidade da frequência cardíaca (Polar), densitometria com dupla emissão de raios X, análise de força muscular em aparelho isocinético e teste cardiopulmonar de exercício submáximo em cicloergômetro. Resultados - Após três meses de intervenção exclusiva com ER observamos uma melhora na vasodilatação endotélio-dependente (P<0,05) e uma redução na pressão arterial sistólica (P<0,01), diastólica (P<0,01) e média (P<0,01), independentes de alterações no peso corporal. Adicionalmente, também observamos redução na frequência cardíaca (FC) de repouso (P<0,01), aumento na variabilidade da FC (P<0,01) e na atividade parasimpática (P<0,05). Observamos também redução na circunferência da cintura (P<0,001), na relação cintura/quadril (P<0,001), no percentual de gordura total (P<0,01), troncular (P<0,01) e de distribuição andróide (P<0,01). Os níveis de fibrinogênio (P<0,05), endotelina-1(P<0,05) e de insulina (P<0,01), e o índice HOMA-IR (P<0,05) foram menores após intervenção. Após a intervenção proposta houve redução na incidência de síndrome metabólica (16,6 vs. 0%) nos adolescentes obesos. Além disso, os estes adolescentes aumentaram a força muscular (P<0,05) e reduziram o consumo de oxigênio, a produção de gás carbônico, a ventilação e o dispêndio energético (P <0,01) durante o teste cardiopulmonar de exercício submáximo. Conclusões - Um treinamento exclusivo com ER resultou em benefícios cardiovasculares, metabólicos e na composição corporal e condicionamento físico de adolescentes obesos, independente de alterações no peso corporal. Nossos resultados sugerem que a prática de ER foi capaz de reduzir fatores de risco cardiovascular em adolescentes com obesidade.

Intravital microscopy evaluation of angiogenesis and its effects on glucose sensor performance.

An optical window model for the rodent dorsum was used to perform chronic and quantitative intravital microscopy and laser Doppler flowmetry of microvascular networks adjacent to functional and non-functional glucose sensors. The one-sided configuration afforded direct, real-time observation of the tissue response to bare (unmodified, smooth surface) sensors and sensors coated with porous poly-L-lactic acid (PLLA). Microvessel length density and red blood cell flux (blood perfusion) within 1 mm of the sensors were measured bi-weekly over 2 weeks. When non-functional sensors were fully implanted beneath the windows, the porous coated sensors had two-fold more vasculature and significantly higher blood perfusion than bare sensors on Day 14. When functional sensors were implanted percutaneously, as in clinical use, no differences in baseline current, neovascularization, or tissue perfusion were observed between bare and porous coated sensors. However, percutaneously implanted bare sensors had two-fold more vascularity than fully implanted bare sensors by Day 14, indicating the other factors, such as micromotion, might be stimulating angiogenesis. Despite increased angiogenesis adjacent to percutaneous sensors, modest sensor current attenuation occurred over 14 days, suggesting that factors other than angiogenesis may play a dominant role in determining sensor function.

Role of nitric oxide in maintenance of basal oral tissue blood flow in anesthetized cats

Experiments were undertaken to determine if nitric oxide (NO) plays a role in regulation of basal blood flow in the oral cavity of pentobarbital anesthetized cats and, if so, to quantify this effect using dose-response relationships. Blood flow was continuously measured from the surface of the tongue and mandibular gingiva (laser-Doppler flowmetry) and from the lingual artery (ultrasonic flowmetry). Cardiovascular parameters also were recorded. Administration of the nonselective inhibitor of nitric oxide synthase (NOS), L-NAME (0.08-20 mg/kg i.v.), produced a dose-related increase of blood pressure associated with decreases of blood flow at all three measurement sites. Maximal blood flow depression of 50-60% was seen 30-60 min after administration of 1.25 mg/kg of L-NAME. D-NAME (1.25 mg/kg i.v.) was inactive at all sites. Subsequent administration of L-arginine partially reversed effects of L-NAME in the lingual artery and tongue, but not in the gingival circulation. The neuronally selective NOS inhibitor, 7-nitroindazole (7-NI, 30 mg/kg i.p.), was devoid of effect on any of the measured parameters. These results suggest that endothelial (but not neuronally derived) NO plays an important role in control of basal blood flow in oral tissues of the cat.

Susceptibility of streptozotocin-induced diabetic rat retinal function and ocular blood flow to acute intraocular pressure challenge.

PURPOSE: To consider whether STZ-induced hyperglycemia renders rat retinal function and ocular blood flow more susceptible to acute intraocular pressure (IOP) challenge.

METHODS: Retinal function (electroretinogram, ERG) was measured during acute IOP challenge (10-100 mmHg, 5 mmHg increments, 3 min/step, vitreal cannulation) in adult Long-Evans rats (6-week old, citrate: n=6, STZ: n=10) 4 weeks after citrate buffer or streptozotocin (STZ, 65 mg/kg, blood glucose > 15 mmol/l) injection. At each IOP, dim and bright flash (-4.56, -1.72 log cd.s.m^-2) ERG responses were recorded to measure inner retinal and ON-bipolar cell function, respectively. Ocular blood flow (laser Doppler flowmetry, citrate; n=6, STZ; n=10) was also measured during acute IOP challenge. Retinae were isolated for qPCR analysis of nitric oxide synthase mRNA expression endothelial, eNos; inducible, iNos; neuronal, nNos).

RESULTS: STZ-induced diabetes increased the susceptibility of inner retinal (IOP at 50% response, 60.1, CI: 57.0-62.0 mmHg vs. citrate: 67.5, CI: 62.1-72.4 mmHg) and ON-bipolar cell function (STZ: 60.3, CI: 58.0-62.8 mmHg vs. citrate: 65.1, CI: 58.0-62.78 mmHg) and ocular blood flow (43.9, CI: 40.8-46.8 vs. citrate: 53.4, CI: 50.7-56.1 mmHg) to IOP challenge. Citrate eyes showed elevated eNos mRNA (+49.7%) after IOP stress, an effect not found in STZ-diabetic eyes (-5.7%, P<0.03). No difference was observed for iNos or nNos (P>0.05) following IOP elevation.

CONCLUSIONS: STZ-induced diabetes increased functional susceptibility during acute IOP challenge. This functional vulnerability is associated with a reduced capacity for diabetic eyes to upregulate eNOS expression and to autoregulate blood flow in response to stress.

Desensitization of thermal hyperemia in the skin is reproducible.

OBJECTIVE: Local heating increases skin blood flow SkBF (thermal hyperemia). In a previous study, we reported that a first local thermal stimulus could attenuate the hyperemic response to a second one applied later on the same skin spot, a phenomenon that we termed desensitization. However, other studies found no evidence for desensitization in similar conditions. The aim of the present work was to test whether it was related to differences in instrumentation. METHODS: Twenty-eight healthy young males were studied. Two pairs of heating chambers, one custom-made (our study) and one commercial (other groups), were affixed to forearm skin. SkBF was measured with single-point laser-Doppler flowmetry (LDF) (780nm) in one pair, and laser-Doppler imaging (LDI) (633nm) in the other. A temperature step from 34 to 41°C, was applied for 30minutes and repeated after two hours. RESULTS: During the second thermal challenge, the plateau SkBF was lower than during the first thermal and was observed with each of the four combinations of SkBF measurement techniques and heating equipment (p<0.05 for all conditions, range -9% to -16% of the initial value). CONCLUSION: Desensitization of thermal hyperemia is not specific to peculiar operating conditions.

Low-Dose Vascular Photodynamic Therapy Decreases Tumor Interstitial Fluid Pressure, which Promotes Liposomal Doxorubicin Distribution in a Murine Sarcoma Metastasis Model.

INTRODUCTION: Solid tumors are known to have an abnormal vasculature that limits the distribution of chemotherapy. We have recently shown that tumor vessel modulation by low-dose photodynamic therapy (L-PDT) could improve the uptake of macromolecular chemotherapeutic agents such as liposomal doxorubicin (Liporubicin) administered subsequently. However, how this occurs is unknown. Convection, the main mechanism for drug transport between the intravascular and extravascular spaces, is mostly related to interstitial fluid pressure (IFP) and tumor blood flow (TBF). Here, we determined the changes of tumor and surrounding lung IFP and TBF before, during, and after vascular L-PDT. We also evaluated the effect of these changes on the distribution of Liporubicin administered intravenously (IV) in a lung sarcoma metastasis model. MATERIALS AND METHODS: A syngeneic methylcholanthrene-induced sarcoma cell line was implanted subpleurally in the lung of Fischer rats. Tumor/surrounding lung IFP and TBF changes induced by L-PDT were determined using the wick-in-needle technique and laser Doppler flowmetry, respectively. The spatial distribution of Liporubicin in tumor and lung tissues following IV drug administration was then assessed in L-PDT-pretreated animals and controls (no L-PDT) by epifluorescence microscopy. RESULTS: L-PDT significantly decreased tumor but not lung IFP compared to controls (no L-PDT) without affecting TBF. These conditions were associated with a significant improvement in Liporubicin distribution in tumor tissues compared to controls (P < .05). DISCUSSION: L-PDT specifically enhanced convection in blood vessels of tumor but not of normal lung tissue, which was associated with a significant improvement of Liporubicin distribution in tumors compared to controls.

Altération du couplage neurovasculaire par l'angiotensine II : évaluation du rôle de la signalisation calcique astrocytaire

Le couplage neurovasculaire (CNV) est un mécanisme d’homéostasie cérébrale régulant le débit sanguin cérébral (CBF) en fonction de l’activité neuronale. La manière dont il est altéré par l’angiotensine II (Ang II), une hormone synthétisée et relâchée dans la circulation systémique ou, alternativement, produite dans le cerveau grâce aux astrocytes, demeure à élucider. Ces cellules expriment le récepteur AT1 (rAT1) et participent à l’orchestration du CNV en relâchant des agents vasoactifs suivant la réponse calcique astrocytaire. Nous avons donc étudié le rôle de cette réponse dans l’altération du CNV induite par l’Ang II. Nous avons trouvé par fluxmétrie par laser Doppler que l’Ang II atténue (p<0.05) la réponse du CBF engendrée par l’activation des récepteurs métabotropes du glutamate du groupe I (mGluRI) du cortex chez la souris C57BL/6. De manière similaire, l’Ang II diminue l'élévation du CBF induite par la stimulation des vibrisses (p<0.05). Sur tranches de cerveaux en aiguë, la polarité de la réponse vasculaire induite par un agoniste mGluRI dans les artérioles parenchymateuses a été significativement renversée par l’Ang II de la vasodilatation vers la vasoconstriction. En parallèle, l’Ang II a augmenté les niveaux de calcium astrocytaire basaux et l’amplitude des réponses calciques (p<0.05). L’altération des réponses vasculaires et calciques maximales a été prévenue par le candesartan, antagoniste des rAT1. Nos résultats suggèrent que l’Ang II potentialise via les rAT1 la réponse calcique qui atteint un seuil favorisant la vasoconstriction par rapport à la vasodilatation, altérant ainsi l’augmentation du CBF en réponse à l’activité neuronale.

A utilização de transformadas de wavelet e de detrended fluctuation analysis no estudo da microcirculação da pele humana

A Fluxometria por Laser Doppler (LDF) é uma técnica não invasiva usada para medir o fluxo microvascular da pele humana. No fluxo é possível isolar componentes oscilatórias em gamas de frequências características que se encontram relacionadas com as actividades cardíaca, respiratória, miogénica, simpática e metabólica. A LDF permite assim estudar a fisiologia do fluxo sanguíneo. Neste trabalho foram realizadas medições de LDF nos tornozelos de 9 mulheres saudáveis numa situação de restrição à perfusão, usando uma braçadeira nos tornozelos. Os dados foram analisados com Transformada de Wavelet e Detrended Fluctuation Analysis (DFA) de modo a estudar os rácios das amplitudes das componentes de Wavelet e os respectivos expoentes . Estes parâmetros foram comparados nas situações de repouso, de restrição à perfusão e de recuperação após remoção da braçadeira. Observou-se que durante a restrição à perfusão houve um aumento significativo dos rácios de amplitude e dos expoentes a para as componentes cardíaca, respiratória e miogénica, o que pode reflectir vasoconstrição. Os parâmetros da componente metabólica apresentaram uma diminuição que se pode relacionar com variações na libertação de NO por parte do endotélio. Após a libertação da braçadeira, os parâmetros das componentes respiratória, miogénica e metabólica retornaram aos valores iniciais. Aanálise combinada de Wavelet com DFAoferece uma nova visão sobre a regulação do fluxo microvascular.

Explorando modelos in vivo para caracterizar a microcirculação periférica : um estudo piloto

Nos últimos anos, a circulação cutânea surgiu como uma janela interessante para analisar a função microcirculatória e os mecanismos de disfunção. Tecnologias não-invasivas, incluindo a Fluxometria por Laser Doppler (FLD), gasimetria transcutânea e a Perda Transepidérmica de Água (PTEA), ajudaram a considerar a circulação cutânea como um modelo de translação útil na doença vascular. Neste estudo procurou-se avaliar o perfil de resposta de um grupo de indivíduos jovens saudáveis ​​(n = 8), de ambos os sexos (24,5 ± 0,8 anos de idade) a três manobras de condicionamento da perfusão no membro inferior - A: elevação da perna enquanto sentado, B: elevação da perna enquanto em decúbito dorsal; C: oclusão supra-sistólica com um torniquete. As técnicas de medição incluiram FLD, pressões parciais transcutâneas (tc) pO2 e pCO2 por gasimetria e PTEA por evaporimetria. Foram aplicados testes de estatística descritiva e não paramétrica, sendo adotado um nível de confiança de 95%. As tcpO2 e tcpCO2 alteraram-se significativamente durante as manobras. Foi registado um perfil de evolução recíproca para FLD e PTEA em A e C, o que pode sugerir que, sob as condições experimentais as condições de perfusão local podem influenciar a função epidérmica "barreira". Os modelos propostos parecem ser adequados para caracterizar a microcirculação periférica in vivo, o que justifica estudos de desenvolvimento posteriores.

Explorando o teste de provocação com oxigénio como modelo de avaliação da microcirculação

A microcirculação cutânea surgiu, nos últimos anos, como uma alternativa pratica e acessível para o estudo da circulação periférica. Técnicas não-invasivas, como a Fluxometria por Laser Doppler (FLD), a Evaporimetria e a Gasimetria transcutânea em associação a testes de provocação têm transformado a circulação cutânea num atraente modelo de investigação. Este estudo foi aplicado a um grupo de voluntárias jovens saudáveis (n = 8, (21,6 ± 2,6) anos) respirando uma atmosfera de 100 % oxigénio durante 10 minutos. Este teste permitiu-nos avaliar a resposta circulatória na microcirculação do membro inferior. As técnicas de medição incluíram o fluxo sanguíneo local por FLD, a pO2 transcutânea (tc) e a Perda Transepidérmica de Água (PTEA) por evaporimetria. A análise de dados revela que tc-pO2 e FLD se alteraram significativamente durante o teste. Um perfil de evolução recíproca foi registrado para FLD e PTEA, que parece apoiar dados anteriores de que as alterações no fluxo sanguíneo local podem influenciar a função de “barreira” epidérmica. Este modelo parece adequado para caracterizar a microcirculação do membro inferior in vivo.

Negative blood oxygen level dependence in the rat: a model for investigating the role of suppression in neurovascular coupling

Modern neuroimaging techniques rely on neurovascular coupling to show regions of increased brain activation. However, little is known of the neurovascular coupling relationships that exist for inhibitory signals. To address this issue directly we developed a preparation to investigate the signal sources of one of these proposed inhibitory neurovascular signals, the negative blood oxygen level-dependent (BOLD) response (NBR), in rat somatosensory cortex. We found a reliable NBR measured in rat somatosensory cortex in response to unilateral electrical whisker stimulation, which was located in deeper cortical layers relative to the positive BOLD response. Separate optical measurements (two-dimensional optical imaging spectroscopy and laser Doppler flowmetry) revealed that the NBR was a result of decreased blood volume and flow and increased levels of deoxyhemoglobin. Neural activity in the NBR region, measured by multichannel electrodes, varied considerably as a function of cortical depth. There was a decrease in neuronal activity in deep cortical laminae. After cessation of whisker stimulation there was a large increase in neural activity above baseline. Both the decrease in neuronal activity and increase above baseline after stimulation cessation correlated well with the simultaneous measurement of blood flow suggesting that the NBR is related to decreases in neural activity in deep cortical layers. Interestingly, the magnitude of the neural decrease was largest in regions showing stimulus-evoked positive BOLD responses. Since a similar type of neural suppression in surround regions was associated with a negative BOLD signal, the increased levels of suppression in positive BOLD regions could importantly moderate the size of the observed BOLD response.

A dynamic model of neurovascular coupling: Implications for blood vessel dilation and constriction

Neurovascular coupling in response to stimulation of the rat barrel cortex was investigated using concurrent multichannel electrophysiology and laser Doppler flowmetry. The data were used to build a linear dynamic model relating neural activity to blood flow. Local field potential time series were subject to current source density analysis, and the time series of a layer IV sink of the barrel cortex was used as the input to the model. The model output was the time series of the changes in regional cerebral blood flow (CBF). We show that this model can provide excellent fit of the CBF responses for stimulus durations of up to 16 s. The structure of the model consisted of two coupled components representing vascular dilation and constriction. The complex temporal characteristics of the CBF time series were reproduced by the relatively simple balance of these two components. We show that the impulse response obtained under the 16-s duration stimulation condition generalised to provide a good prediction to the data from the shorter duration stimulation conditions. Furthermore, by optimising three out of the total of nine model parameters, the variability in the data can be well accounted for over a wide range of stimulus conditions. By establishing linearity, classic system analysis methods can be used to generate and explore a range of equivalent model structures (e.g., feed-forward or feedback) to guide the experimental investigation of the control of vascular dilation and constriction following stimulation.

A three-compartment model of the hemodynamic response and oxygen delivery to brain

We describe a mathematical model linking changes in cerebral blood flow, blood volume and the blood oxygenation state in response to stimulation. The model has three compartments to take into account the fact that the cerebral blood flow and volume as measured concurrently using laser Doppler flowmetry and optical imaging spectroscopy have contributions from the arterial, capillary as well as the venous compartments of the vasculature. It is an extension to previous one-compartment hemodynamic models which assume that the measured blood volume changes are from the venous compartment only. An important assumption of the model is that the tissue oxygen concentration is a time varying state variable of the system and is driven by the changes in metabolic demand resulting from changes in neural activity. The model takes into account the pre-capillary oxygen diffusion by flexibly allowing the saturation of the arterial compartment to be less than unity. Simulations are used to explore the sensitivity of the model and to optimise the parameters for experimental data. We conclude that the three-compartment model was better than the one-compartment model at capturing the hemodynamics of the response to changes in neural activation following stimulation.

Long duration stimuli and nonlinearities in the neural–haemodynamic coupling

Recent studies have shown that the haemodynamic responses to brief (<2 secs) stimuli can be well characterised as a linear convolution of neural activity with a suitable haemodynamic impulse response. In this paper, we show that the linear convolution model cannot predict measurements of blood flow responses to stimuli of longer duration (>2 secs), regardless of the impulse response function chosen. Modifying the linear convolution scheme to a nonlinear convolution scheme was found to provide a good prediction of the observed data. Whereas several studies have found a nonlinear coupling between stimulus input and blood flow responses, the current modelling scheme uses neural activity as an input, and thus implies nonlinearity in the coupling between neural activity and blood flow responses. Neural activity was assessed by current source density analysis of depth-resolved evoked field potentials, while blood flow responses were measured using laser Doppler flowmetry. All measurements were made in rat whisker barrel cortex after electrical stimulation of the whisker pad for 1 to 16 secs at 5 Hz and 1.2 mA (individual pulse width 0.3 ms).

The hemodynamic impulse response to a single neural event

This article investigates the relation between stimulus-evoked neural activity and cerebral hemodynamics. Specifically, the hypothesis is tested that hemodynamic responses can be modeled as a linear convolution of experimentally obtained measures of neural activity with a suitable hemodynamic impulse response function. To obtain a range of neural and hemodynamic responses, rat whisker pad was stimulated using brief (less than or equal to2 seconds) electrical stimuli consisting of single pulses (0.3 millisecond, 1.2 mA) combined both at different frequencies and in a paired-pulse design. Hemodynamic responses were measured using concurrent optical imaging spectroscopy and laser Doppler flowmetry, whereas neural responses were assessed through current source density analysis of multielectrode recordings from a single barrel. General linear modeling was used to deconvolve the hemodynamic impulse response to a single "neural event" from the hemodynamic and neural responses to stimulation. The model provided an excellent fit to the empirical data. The implications of these results for modeling schemes and for physiologic systems coupling neural and hemodynamic activity are discussed.