Efeito do betacaroteno sobre o estresse oxidativo e a expressão de conexina 43 cardíaca / Effect of beta-carotene on oxidative stress and expression of cardiac connexin 43

FUNDAMENTO: Estudos de intervenção mostraram aumento da mortalidade em pacientes que receberam betacaroteno. Contudo, não são conhecidos os mecanismos envolvidos nesse fenômeno. OBJETIVO: Avaliar a influência do betacaroteno sobre o estresse oxidativo e a expressão de conexina 43 em coração de ratos. MÉTODOS: Ratos Wistar, pesando aproximadamente 100 g, foram alocados em dois grupos: Grupo Controle (n = 30), que recebeu a dieta usada de rotina em nosso laboratório, e Grupo Betacaroteno (n = 28), que recebeu betacaroteno (na forma de cristal, adicionado e misturado à dieta) na dose de 500 mg de betacaroteno/kg de dieta. Os animais receberam tratamento até que atingissem entre 200 e 250 g, quando eram sacrificados. Foram coletados sangue, fígado e coração para realização de Western blotting e imunoistoquímica para conexina 43; foram realizados estudos morfométricos, dosagens de betacaroteno por cromatografia líquida de alta eficiência bem como de glutationa reduzida, glutationa oxidada e hidroperóxidos de lipídeos por análises bioquímicas. RESULTADOS: O betacaroteno foi detectado apenas no fígado dos animais do Grupo Betacaroteno (288 ± 94,7 µg/kg). Os níveis de glutationa reduzida/glutationa oxidada foram maiores no fígado e no coração dos animais do Grupo Betacaroteno (fígado - Grupo Controle: 42,60 ± 1,62; fígado - Grupo Betacaroteno: 57,40 ± 5,90; p = 0,04; coração: - Grupo Controle: 117,40 ± 1,01; coração - Grupo Betacaroteno: 121,81 ± 1,32 nmol/mg proteína; p = 0,03). O conteúdo de conexina 43 total foi maior no Grupo Betacaroteno. CONCLUSÃO: O betacaroteno apresentou efeito benéfico, caracterizado pelo aumento da comunicação intercelular e melhora do sistema de defesa antioxidante. Nesse modelo, os mecanismos não explicam a maior mortalidade observada com a suplementação de betacaroteno em estudos clínicos. (Arq Bras Cardiol. 2013; [online].ahead print, PP.0-0).

BACKGROUND: Intervention studies have shown an increased mortality in patients who received beta-carotene. However, the mechanisms involved in this phenomenon are still unknown. OBJECTIVE: Evaluate the influence of beta-carotene on oxidative stress and the expression of connexin 43 in rat hearts. METHODS: Wistar rats, weighing approximately 100 g, were allocated in two groups: Control Group (n=30), that received the diet routinely used in our laboratory, and Beta-Carotene Group (n = 28), which received beta-carotene (in crystal form, added and mixed to the diet) at a dose of 500 mg of beta-carotene/kg of diet. The animals received the treatment until they reached 200-250g, when they were sacrificed. Samples of blood, liver and heart were collected to perform Western blotting and immunohistochemistry for connexin 43; morphometric studies, dosages of beta-carotene by high-performance liquid chromatography as well as reduced glutathione, oxidized glutathione and lipids hydroperoxides were performed by biochemical analysis. RESULTS: Beta-carotene was detected only in the liver of Beta-Carotene Group animals (288 ± 94.7 µg/kg). Levels of reduced/oxidized glutathione were higher in the liver and heart of Beta-Carotene Group animals (liver - Control Group: 42.60 ± 1.62; liver - Beta-Carotene Group: 57.40 ± 5.90; p = 0.04; heart: - Control Group: 117.40 ± 1.01; heart - Beta-Carotene Group: 121.81 ± 1.32 nmol/mg protein; p = 0.03). The content of total connexin 43 was larger in Beta-Carotene Group. CONCLUSION: Beta-carotene demonstrated a positive effect, characterized by the increase of intercellular communication and improvement of anti-oxidizing defense system. In this model, mechanism does not explain the increased mortality rate observed with the beta-carotene supplementation in clinical studies. (Arq Bras Cardiol. 2013; [online].ahead print, PP.0-0).

Effect of beta-carotene on oxidative stress and expression of cardiac connexin 43.

BACKGROUND: Intervention studies have shown an increased mortality in patients who received beta-carotene. However, the mechanisms involved in this phenomenon are still unknown. OBJECTIVE: Evaluate the influence of beta-carotene on oxidative stress and the expression of connexin 43 in rat hearts. METHODS: Wistar rats, weighing approximately 100 g, were allocated in two groups: CONTROL GROUP (n=30), that received the diet routinely used in our laboratory, and Beta-Carotene Group (n = 28), which received beta-carotene (in crystal form, added and mixed to the diet) at a dose of 500 mg of beta-carotene/kg of diet. The animals received the treatment until they reached 200-250 g, when they were sacrificed. Samples of blood, liver and heart were collected to perform Western blotting and immunohistochemistry for connexin 43; morphometric studies, dosages of beta-carotene by high-performance liquid chromatography as well as reduced glutathione, oxidized glutathione and lipids hydroperoxides were performed by biochemical analysis. RESULTS: Beta-carotene was detected only in the liver of Beta-Carotene Group animals (288 ± 94.7 µg/kg). Levels of reduced/oxidized glutathione were higher in the liver and heart of Beta-Carotene Group animals (liver - CONTROL GROUP: 42.60 ± 1.62; liver - Beta-Carotene Group: 57.40 ± 5.90; p = 0.04; heart: - CONTROL GROUP: 117.40 ± 1.01; heart - Beta-Carotene Group: 121.81 ± 1.32 nmol/mg protein; p = 0.03). The content of total connexin 43 was larger in Beta-Carotene Group. CONCLUSION: Beta-carotene demonstrated a positive effect, characterized by the increase of intercellular communication and improvement of anti-oxidizing defense system. In this model, mechanism does not explain the increased mortality rate observed with the beta-carotene supplementation in clinical studies.

Smoking is associated with remodeling of gap junction in the rat heart: smoker's paradox explanation?

BACKGROUND: In a previous study utilizing the rat model, exposure to tobacco smoke for 5 weeks increased survival after AMI, despite similar age and infarct size between the smokers and nonsmokers, and absence of reperfusion. OBJECTIVE: Thus, this study aimed to analyze the effects of exposure to tobacco smoke on intensity, distribution or phosphorylation of connexin 43 in the rat heart. METHODS: Wistar rats weighing 100 g were randomly allocated into 2 groups: 1) CONTROL (n = 25); 2) Exposed to tobacco smoke (ETS), n = 23. After 5 weeks, left ventricular morphometric analysis, immunohistochemistry and western blotting for connexin 43 (Cx43) were performed. RESULTS: Collagen volume fraction, cross-sectional areas, and ventricular weight were not statistically different between control and ETS. ETS showed lower stain intensity of Cx43 at intercalated disks ( CONTROL: 2.32 ± 0.19; ETS: 1.73 ± 0.18; p = 0.04). The distribution of CX43 at intercalated disks did not differ between the groups ( CONTROL: 3.73 ± 0.12; ETS: 3.20 ± 0.17; p = 0.18). ETS rats showed higher levels of dephosphorylated form of Cx43 (CONTROL: 0.45 ± 0.11; ETS: 0.90 ± 0.11; p = 0.03). On the other hand, total Cx43 did not differ between control and ETS groups ( CONTROL: 0.75 ± 0.19; ETS: 0.93 ± 0.27; p = 0.58). CONCLUSION: Exposure to tobacco smoke resulted in cardiac gap junction remodeling, characterized by alterations in the quantity and phosphorylation of the Cx43, in rats hearts. This finding could explain the smoker's paradox observed in some studies.

O tabagismo está associado com a remodelação de junções comunicantes no coração de ratos: explicação do paradoxo dos fumantes? / Smoking is associated with remodeling of gap junction in the rat heart: smoker's paradox explanation?

FUNDAMENTO: Em estudo anterior, utilizando o modelo de ratos, a exposição à fumaça do cigarro durante 5 semanas aumentou a sobrevida após IAM, apesar da idade similar e tamanho do infarto entre fumantes e não fumantes, e da ausência de reperfusão. OBJETIVO: Dessa forma, o presente estudo teve como objetivo analisar os efeitos da exposição à fumaça do cigarro sobre a intensidade, distribuição ou fosforilação da conexina 43 no coração de ratos. MÉTODOS: Ratos Wistar, pesando 100 g, foram distribuídos aleatoriamente em 2 grupos: 1) Controle (n = 25); 2) Expostos à fumaça do cigarro (ETS), n = 23. Depois de 5 semanas, foram conduzidas análise morfométrica do ventrículo esquerdo, imuno-histoquímica e Western blot para conexina 43 (Cx43). RESULTADOS: A fração do volume de colágeno, as áreas transversais e o peso ventricular não foram estatisticamente diferentes entre os grupos controle e ETS. O grupo ETS apresentou uma coloração de menor intensidade da Cx43 em discos intercalados (Controle: 2,32 ± 0,19; ETS: 1,73 ± 0,18; p = 0,04). A distribuição da Cx43 em discos intercalados não diferiu entre os grupos (Controle: 3,73 ± 0,12; ETS: 3,20 ± 0,17; p = 0,18). Os ratos do grupo ETS mostraram um nível maior de forma desfosforilada da Cx43 (Controle: 0,45 ± 0,11; ETS: 0,90 ± 0,11; p = 0,03). Por outro lado, o Cx43 total não diferiu entre os grupos de controle e ETS (Controle: 0,75 ± 0,19; ETS: 0,93 ± 0,27; p = 0,58). CONCLUSÃO: A exposição à fumaça do cigarro resultou na remodelação das junções comunicantes cardíacas, caracterizada por alterações na quantidade e fosforilação da Cx43 em corações de ratos. Essa constatação pode explicar o paradoxo dos fumantes observado em alguns estudos.

BACKGROUND: In a previous study utilizing the rat model, exposure to tobacco smoke for 5 weeks increased survival after AMI, despite similar age and infarct size between the smokers and nonsmokers, and absence of reperfusion. OBJECTIVE: Thus, this study aimed to analyze the effects of exposure to tobacco smoke on intensity, distribution or phosphorylation of connexin 43 in the rat heart. METHODS: Wistar rats weighing 100 g were randomly allocated into 2 groups: 1) Control (n = 25); 2) Exposed to tobacco smoke (ETS), n = 23. After 5 weeks, left ventricular morphometric analysis, immunohisthochemistry and western blotting for connexin 43 (Cx43) were performed. RESULTS: Collagen volume fraction, cross-sectional areas, and ventricular weight were not statistically different between control and ETS. ETS showed lower stain intensity of Cx43 at intercalated disks (Control: 2.32 ± 0.19; ETS: 1.73 ± 0.18; p = 0.04). The distribution of CX43 at intercalated disks did not differ between the groups (Control: 3.73 ± 0.12; ETS: 3.20 ± 0.17; p = 0.18). ETS rats showed higher levels of dephosphorylated form of Cx43 (Control: 0.45 ± 0.11; ETS: 0.90 ± 0.11; p = 0.03). On the other hand, total Cx43 did not differ between control and ETS groups (Control: 0.75 ± 0.19; ETS: 0.93 ± 0.27; p = 0.58). CONCLUSION: Exposure to tobacco smoke resulted in cardiac gap junction remodeling, characterized by alterations in the quantity and phosphorylation of the Cx43, in rats hearts. This finding could explain the smoker's paradox observed in some studies.

Influence of AIN-93 diet on mortality and cardiac remodeling after myocardial infarction in rats.

BACKGROUND: The AIN-93 diet was proposed by the American Institute of Nutrition with the objective of standardising studies in experimental nutrition. Our objective was to analyze the effects of AIN-93 diet after myocardial infarction in rats. METHODS: Post weaning, the animals were divided into two groups: control (C, n=62), fed the standard diet of our laboratory (Labina); AIN-93 Group (n=70), fed the AIN-93 diet. Achieving 250 g, the animals were subjected to myocardial infarction. RESULTS: Early mortality was increased in AIN-93 animals, associated with lower serum levels of calcium, magnesium, potassium, sodium, and phosphorus. On the other hand, after 90 days, AIN-93 showed smaller normalized left ventricular dimensions. The caloric and carbohydrate intake was smaller, but the fat intake was higher in AIN-93 rats. AIN-93 group also showed increased levels of ß-hydroxyacylcoenzyme A dehydrogenase and citrate synthase. In addition, serum levels of insulin and cardiac levels of malondialdehyde, metalloproteinases-2 and -9, and TNF-α and IFN-γ were decreased in the AIN-93 group. CONCLUSION: AIN-93 diet increased early mortality, while attenuated the chronic remodeling process after experimental coronary occlusion. Therefore, this diet has biological effects and should be use with attention in this model.

Influence of different doses of retinoic acid on cardiac remodeling.

OBJECTIVE: The role of retinoic acid in promoting postnatal heart alterations is still unclear. The aim of this study was to evaluate whether the cardiac alterations caused by all-trans- retinoic acid (ATRA) in normal adult rat hearts are physiologic or pathologic and if these alterations are dose-dependent. METHODS: Rats were allocated into a control group that received a diet without ATRA (n=16), a group that received 0.3 mg of ATRA/kg of diet (n=17), a group that received a diet containing 10 mg of ATRA/kg (n=18), or a group that received 50 mg of ATRA/kg in the diet (n=18). After 4 wk, the animals were evaluated echocardiographically, morphologically, and biochemically. RESULTS: The 50-mg ATRA group presented cardiac hypertrophy with maintenance of cardiac geometry and increased systolic function, whereas diastolic function was similar to that of the control group. In addition, progressive increases in the ATRA dose resulted in gradual augmentations of left atrial diameter, left ventricular diastolic and systolic diameters, left ventricular mass index, cardiac output, cardiac index, and aortic velocity. The ATRA did not produce alterations in interferon-γ and tumor necrosis factor-α cardiac levels, interstitial collagen volume fraction, or the intensity and localization of connexin-43. In addition, no alteration was observed in ß-hydroxyacyl coenzyme A dehydrogenase, lactate dehydrogenase, or citrate synthase, suggesting that cardiac energetic metabolism was preserved with ATRA. CONCLUSION: These results suggest that ATRA produced dose-dependent effects and cardiac remodeling that is more compatible with a physiologic response.

Role of lipoperoxidation in the remodeling intensification induced by beta-carotene after infarction.

BACKGROUND: The mechanisms involved in the biggest remodeling caused by the post-infarct beta-carotene are unknown. OBJECTIVE: To analyze the role of lipoperoxidation in the ventricular remodeling after infarct of the myocardium in rats supplemented with beta-carotene. METHODS: Rats were infarcted and divided into two groups: C (control) and BC (500mg/kg/regimen). After six months, echocardiogram and biochemical evaluation were performed. The t test was used, with 5% significance. RESULTS: The animals from BC group presented highest means of the diastolic (C = 1.57 +/- 0.4 mm(2)/g, BC = 2.09 +/- 0.3 mm(2)/g; p < 0.001) and systolic (C = 1.05 +/- 0.3 mm(2)/g, BC = 1.61 +/- 0.3 mm(2)/g; p < 0.001) areas of LV, which were adapted according to the rat's body weight. The systolic function of LV, evaluated by the area variation fraction, was lower in the animals supplemented with beta-carotene (C = 31.9 +/- 9.3%, BC = 23.6 +/- 5.1%; p = 0.006). The animals supplemented with beta-carotene presented higher values of the E/A relation (C = 2.7 +/- 2.5, BC = 5.1 +/- 2.8; p = 0.036). No differences were found between the groups concerning the cardiac levels of the GSH (C = 21 +/- 8 nmol/mg of protein, BC = 37 +/- 15 nmol/mg of protein; p = 0.086), GSSG (C = 0.4 (0.3-0.5) nmol/g of protein, BC = 0.8 (0.4-1.0; p = 0.19) of protein; p = 0.246) and lipoperoxides (C = 0.4 +/- 0.2 nmol/mg of tissue, BC = 0.2 +/- 0.1 nmol/mg of tissue; p = 0.086). CONCLUSION: The highest remodeling in infarcted rats supplemented with beta-carotene does not depend on the lipoperoxidation.

Papel da lipoperoxidação na intensificação da remodelação causada pelo betacaroteno após o infarto / Rol de la lipoperoxidación en la intensificación de la remodelación ocasionada por el betacaroteno tras infarto / Role of lipoperoxidation in the remodeling intensification induced by beta-carotene after infarction

FUNDAMENTO: Os mecanismos envolvidos na maior remodelação causada pelo betacaroteno após o infarto são desconhecidos. OBJETIVO: Analisar o papel da lipoperoxidação na remodelação ventricular após o infarto do miocárdio, em ratos suplementados com betacaroteno. MÉTODOS: Ratos foram infartados e distribuídos em dois grupos: C (controle) e BC (500mg/kg/dieta). Após seis meses, foram realizados ecocardiograma e avaliação bioquímica. Utilizamos o teste t, com significância de 5 por cento. RESULTADOS: Os animais do grupo BC apresentaram maiores médias das áreas diastólicas (C = 1,57 ± 0,4 mm²/g, BC = 2,09 ± 0,3 mm²/g; p < 0,001) e sistólicas (C = 1,05 ± 0,3 mm²/g, BC = 1,61 ± 0,3 mm²/g; p < 0,001) do VE, ajustadas ao peso corporal do rato. A função sistólica do VE, avaliada pela fração de variação de área, foi menor nos animais suplementados com betacaroteno (C = 31,9 ± 9,3 por cento, BC = 23,6 ± 5,1 por cento; p = 0,006). Os animais suplementados com betacaroteno apresentaram valores maiores da relação E/A (C = 2,7 ± 2,5, BC = 5,1 ± 2,8; p = 0,036). Não foram encontradas diferenças entre os grupos em relação aos níveis cardíacos de GSH (C = 21 ± 8 nmol/mg de proteína, BC = 37 ±15 nmol/mg de proteína; p = 0,086), GSSG (C = 0,4 (0,3-0,5) nmol/g de proteína, BC = 0,8 (0,4-1,0; p = 0,19) de proteína; p = 0,246) e lipoperóxidos (C = 0,4 ± 0,2 nmol/mg de tecido, BC = 0,2 ± 0,1 nmol/mg de tecido; p = 0,086). CONCLUSÃO: A maior remodelação em animais infartados e suplementados com betacaroteno não depende da lipoperoxidação.

BACKGROUND: The mechanisms involved in the biggest remodeling caused by the post-infarct beta-carotene are unknown. OBJECTIVE: To analyze the role of lipoperoxidation in the ventricular remodeling after infarct of the myocardium in rats supplemented with beta-carotene. METHODS: Rats were infarcted and divided into two groups: C (control) and BC (500mg/kg/regimen). After six months, echocardiogram and biochemical evaluation were performed. The t test was used, with 5 percent significance. RESULTS: The animals from BC group presented highest means of the diastolic (C = 1.57 ± 0.4 mm²/g, BC = 2.09 ± 0.3 mm²/g; p < 0.001) and systolic (C = 1.05 ± 0.3 mm²/g, BC = 1.61 ± 0.3 mm²/g; p < 0.001) areas of LV, which were adapted according to the rat's body weight. The systolic function of LV, evaluated by the area variation fraction, was lower in the animals supplemented with beta-carotene (C = 31.9 ± 9.3 percent, BC = 23.6 ± 5.1 percent; p = 0.006). The animals supplemented with beta-carotene presented higher values of the E/A relation (C = 2.7 ± 2.5, BC = 5.1 ± 2.8; p = 0.036). No differences were found between the groups concerning the cardiac levels of the GSH (C = 21 ± 8 nmol/mg of protein, BC = 37 ± 15 nmol/mg of protein; p = 0.086), GSSG (C = 0.4 (0.3-0.5) nmol/g of protein, BC = 0.8 (0.4-1.0; p = 0.19) of protein; p = 0.246) and lipoperoxides (C = 0.4 ± 0.2 nmol/mg of tissue, BC = 0.2 ± 0.1 nmol/mg of tissue; p = 0.086). CONCLUSION: The highest remodeling in infarcted rats supplemented with beta-carotene does not depend on the lipoperoxidation.

FUNDAMENTO: Los mecanismos implicados en la mayor remodelación ocasionada por betacaroteno tras el infarto son desconocidos. OBJETIVO: Analizar el rol que juega la lipoperoxidación en la remodelación ventricular tras el infarto de miocardio, en ratas suplementadas con betacaroteno. MÉTODOS: Se había inducido a un infarto a las ratas y se las distribuyó en grupos: C (control) y BC (500mg/kg/dieta). Tras seis meses, se realizaron ecocardiograma y evaluación bioquímica. Utilizamos la prueba t, con significancia del 5 por ciento. RESULTADOS: Los animales del grupo BC presentaron mayores promedios de las áreas diastólicas (C = 1,57 ± 0,4 mm²/g, BC = 2,09 ± 0,3 mm²/g; p < 0,001) y sistólicas (C = 1,05 ± 0,3 mm²/g, BC = 1,61 ± 0,3 mm²/g; p < 0,001) del VI, ajustadas al peso corporal de la rata. La función sistólica del VI, evaluada por la fracción de variación de área, fue menor en los animales suplementados con betacaroteno (C = 31,9 ± 9,3 por ciento, BC = 23,6 ± 5,1 por ciento; p = 0,006). Los animales suplementados con betacaroteno presentaron valores mayores de la relación E/A (C = 2,7 ± 2,5, BC = 5,1 ± 2,8; p = 0,036). No se encontraron diferencias entre los grupos con relación a los niveles cardiacos de GSH (C = 21 ± 8 nmol/mg de proteína, BC = 37 ±15 nmol/mg de proteína; p = 0,086), GSSG (C = 0,4 (0,3-0,5) nmol/g de proteína, BC = 0,8 (0,4-1,0; p = 0,19) de proteína; p = 0,246) y lipoperóxidos (C = 0,4 ± 0,2 nmol/mg de tejido, BC = 0,2 ± 0,1 nmol/mg de tejido; p = 0,086). CONCLUSIÓN: La mayor remodelación en animales infartados y suplementados con betacaroteno no depende de la lipoperoxidación.

Influência da exposição à fumaça de cigarro e da suplementação da dieta com Beta-Caroteno sobre a comunicação intercelular em cardiomiócitos de ratos / Influence of exposure to cigarette smoke and supplementation of diet with beta-carotene on the intercellular communication in cardiomyocytes of rates

Em estudo anterior realizado em nosso laboratório ratos expostos à fumaça de cigarro apresentaram menor mortalidade após infarto agudo do miocárdio em relação ao grupo controle. Entretanto a mortalidade aumentou nos ratos fumantes quando receberam Beta-caroteno. Não se sabe qual o mecanismo da diminuição de mortalidade nos animais fumantes. Formulamos a hipótese de que a fumaça de cigarro induziria estresse oxidativo que causaria efeito tipo precondicionamento cardíaco. O efeito tipo precondicionamento diminuiria a comunicação intercelular pelas junções comunicantes. A suplementação da dieta com Beta-caroteno inibiria os efeitos provenientes da exposição à fumaça de cigarro. Foram estudados ratos Wistar, distribuídos em 4 grupos: grupo controle (C) - formado por 30 animais, que receberam a dieta usada de rotina em nosso laboratório; 2) grupo Beta-caroteno (SC) - composto por 28 animais que receberam Beta-caroteno (na forma de cristal adicionado e misturado à dieta) na dose de 500 mg de Beta-caroteno/kg de dieta; 3) grupo exposto à fumaça de cigarro (EFC) composto por 28 animais, que receberam a dieta usada de rotina em nosso laboratório e foram expostos ao cigarro; 4) grupo Beta-caroteno/EFC (SC/EFC) - formado por 28 animais que receberam o Beta-caroteno na mesma forma que o grupo SC e foram expostos à fumaça de cigarro. A suplementação com Beta-­caroteno e a EFC foram realizados simultaneamente, até que ratos, com peso inicial de 100g, atingissem peso corpóreo de 200-250g...

Exposure to stress differential vascular adaptive response in spontaneously hypertensive and Wistar rats: Role of nitric oxide, and prehypertensive and hypertensive states.

UNLABELLED: Stress-induced vascular adaptive response in SHR was investigated, focusing on the endothelium. Noradrenaline responses were studied in intact and denuded aortas from 6-week-old (prehypertensive) and 14-week-old (hypertensive) SHR and age-matched Wistar rats submitted or not to acute stress (20-min swimming and 1-h immobilization 25 min apart), preceded or not by chronic stress (2 sessions 2 days apart of 1-h day immobilization for 5-consecutive days). Stress did not alter the reactivity of denuded aorta. Moreover, no alteration in the EC50 values was observed after stress exposure. In intact aortas, acute stress-induced hyporeactivity to noradrenaline similar between strains at both age. Chronic stress potentiated this adaptive response in 6- and 14-week-old Wistar but not in 6-week-old SHR, and did not alter the reactivity of 14-week-old SHR. Maximum response (g) in intact aortas [6-week-old: Wistar 3.25+/-0.12, Wistar/acute 1.95+/-0.12*, Wistar/chronic 1.36+/-0.21*(+), SHR 1.75+/-0.11, SHR/acute 0.88+/-0.08*, SHR/chronic 0.85+/-0.05*; 14-week-old: Wistar 3.83+/-0.13, Wistar/acute 2.72+/-0.13*, Wistar/chronic 1.91+/-0.19*(+), SHR 4.03+/-0.17, SHR/acute 2.26+/-0.12*, SHR/chronic 4.10+/-0.23; inside the same strain: *P < 0.05 relate to non-stressed rat, +P < 0.05 related to acute stressed rat; n = 6-18]. Independent of age and strain, L-NAME and endothelium removal abolished the stress-induced aorta hyporeactivity. CONCLUSION: The vascular adaptive response to stress is impaired in SHR, independently of the hypertensive state. Moreover, this vascular adaptive response is characterized by endothelial nitric oxide-system hyperactivity in both strains.

Beta-carotene supplementation attenuates cardiac remodeling induced by one-month tobacco-smoke exposure in rats.

OBJECTIVE: The objectives were to analyze the cardiac effects of exposure to tobacco smoke (ETS), for a period of 30 days, alone and in combination with beta-carotene supplementation (BC). RESEARCH METHODS AND PROCEDURES: Rats were allocated into: Air (control, n = 13); Air + BC (n = 11); ETS (n = 11); and BC + ETS (n = 9). In Air + BC and BC + ETS, 500 mg of BC were added to the diet. After three months of randomization, cardiac structure and function were assessed by echocardiogram. After that, animals were euthanized and morphological data were analyzed post-mortem. One-way and two-way ANOVA were used to assess the effects of ETS, BC and the interaction between ETS and BC on the variables. RESULTS: ETS presented smaller cardiac output (0.087 +/- 0.001 vs. 0.105 +/- 0.004 l/min; p = 0.007), higher left ventricular diastolic diameter (19.6 +/- 0.5 vs. 18.0 +/- 0.5 mm/kg; p = 0.024), higher left ventricular (2.02 +/- 0.05 vs. 1.70 +/- 0.03 g/kg; p < 0.001) and atrium (0.24 +/- 0.01 vs. 0.19 +/- 0.01 g/kg; p = 0.003) weight, adjusted to body weight of animals, and higher values of hepatic lipid hydroperoxide (5.32 +/- 0.1 vs. 4.84 +/- 0.1 nmol/g tissue; p = 0.031) than Air. However, considering those variables, there were no differences between Air and BC + ETS (0.099 +/- 0.004 l/min; 19.0 +/- 0.5 mm/kg; 1.83 +/- 0.04 g/kg; 0.19 +/- 0.01 g/kg; 4.88 +/- 0.1 nmol/g tissue, respectively; p > 0.05). Ultrastructural alterations were found in ETS: disorganization or loss of myofilaments, plasmatic membrane infolding, sarcoplasm reticulum dilatation, polymorphic mitochondria with swelling and decreased cristae. In BC + ETS, most fibers showed normal morphological aspects. CONCLUSION: One-month tobacco-smoke exposure induces functional and morphological cardiac alterations and BC supplementation attenuates this ventricular remodeling process.

beta-carotene attenuates the paradoxical effect of tobacco smoke on the mortality of rats after experimental myocardial infarction.

The objective of this study was to investigate the effects of exposure to tobacco smoke (ETS) in rats that were or were not supplemented with dietary beta-carotene (BC), on ventricular remodeling and survival after myocardial infarction (MI). Rats (n = 189) were allocated to 4 groups: the control group, n = 45; group BC administered 500 mg/kg diet, n = 49, BC supplemented rats; group ETS, n = 55, rats exposed to tobacco smoke; and group BC+ETS, n = 40. Wistar rats weighing 100 g were administered one of the treatments until they weighed 200 to 250 g (approximately 5 wk). The ETS rats were exposed to cigarette smoke for 30 min 4 times/d, in a chamber connected to a smoking device. After reaching a weight of 200-250 g, rats were subjected to experimental MI (coronary artery occlusion) and mortality rates were determined over the next 105 d. In addition, echocardiographic, isolated heart, morphometrical, and biochemical studies were performed. Mortality data were tested using Kaplan-Meyer curves and other data by 2-way ANOVA. Survival rates were greater in the ETS group (58.2%) than in the control (33.3%) (P = 0.001) and BC+ETS rats (30.0%) (P = 0.007). The groups did not differ in the other comparisons. Left ventricular end-diastolic diameter normalized to body weight was greater and maximal systolic pressures were lower in the ETS groups than in non-ETS groups. Previous exposure to tobacco smoke induced a process of cardiac remodeling after MI. There is a paradoxical protector effect with tobacco smoke exposure, characterized by lower mortality, which is offset by BC supplementation.