RESUMO
Epidemiological studies suggest a positive association between nutrient intake, hyperinsulinemia and risk of Benign prostatic hyperplasis (BPH). This study tests the hypothesis that a low-fat, high-fiber diet and daily exercise would lower serum insulin and reduce the growth of serum-stimulated primary prostate epithelial cells in culture. Serum samples were obtained from eight overweight men before and after the Pritikin residential, 2-week diet and exercise intervention and from seven men who were long-term followers of the low-fat, high-fiber diet and regular exercise lifestyle. The serum was used to stimulate primary prostate epithelial cells in culture. Growth was measured after 48 and 96 h and apoptosis after 96 h. At 48 h there was no significant difference in growth within the Pre, 2-week or Long-Term groups. At 96 h growth was significantly reduced in the 2-week (13%) and in the Long-Term (14%) groups compared to the Pre data. At 96 h, apoptosis was not significantly different among the three groups. Fasting insulin was reduced by 30% in the 2-week group and by 52% in the Long-Term group compared to the Pre data. Testosterone was unchanged in the 2-week group. The results of this study indicate that a low-fat, high-fiber diet and daily exercise lowers insulin and reduces growth of prostate primary epithelial cells and suggests that lifestyle may be an important factor in the development or progression of BPH. Future prospective trials should address the effects of this lifestyle modification on BPH symptomatology and progression.
Assuntos
Dieta , Exercício Físico , Neoplasias da Próstata/patologia , Proliferação de Células , Células Cultivadas , Células Epiteliais/citologia , Humanos , Resistência à Insulina , Masculino , Pessoa de Meia-Idade , Neoplasias da Próstata/sangueRESUMO
For over 40 years, avian sarcoma and leukosis virus (ASLV)-receptor interactions have been employed as a useful model system to study the mechanism of retroviral entry into cells. Pioneering studies on this system focused upon the genetic basis of the differential susceptibilities of different lines of chickens to infection by distinct subgroups of ASLV. These studies led to the definition of three distinct autosomal recessive genes that were predicted to encode cellular receptors for different viral subgroups. They also led to the concept of viral interference, i.e. the mechanism by which infection by one virus can render cells resistant to reinfection by other viruses that use the same cellular receptor. Here, we review the contributions that analyses of the ASLV-receptor system have made in unraveling the mechanisms of retroviral entry into cells and focus on key findings such as identification and characterization of the ASLV receptor genes and the subsequent elucidation of an unprecedented mechanism of virus-cell fusion. Since many of the initial findings on this system were published in the early volumes of Virology, this subject is especially well suited to this special anniversary issue of the journal.
Assuntos
Vírus da Leucose Aviária/metabolismo , Receptores Virais/metabolismo , Sarcoma Aviário/virologia , Proteínas Virais de Fusão/metabolismo , Animais , Vírus da Leucose Aviária/fisiologia , Fusão de Membrana , Receptores Virais/genética , Proteínas Virais/química , Proteínas Virais/metabolismo , Proteínas Virais/fisiologia , Replicação ViralRESUMO
Binding of avian sarcoma and leukosis virus (ASLV) to its cognate receptor on the cell surface causes conformational changes in its envelope protein (Env). It is currently debated whether low pH is required for ASLV infection. To elucidate the role of low pH, we studied the association between ASLV subgroup B (ASLV-B) and liposomes and fusion between effector cells expressing Env from ASLV-A and ASLV-B and target cells expressing cognate receptors. Neither EnvA nor EnvB promoted cell-cell fusion at neutral pH, but lowering the pH resulted in quick and extensive fusion. As expected for a low-pH-triggered reaction, fusion was a steep function of pH. Steps that required low pH were identified. Binding a soluble form of the receptor caused ASLV-B to hydrophobically associate with liposome membranes at neutral pH, indicating that low pH is not required for insertion of Env's fusion peptides into membranes. But both cell-cell hemifusion and fusion pore formation were pH dependent. It is proposed that fusion peptide insertion stabilizes the conformation of ASLV Env into a form that can be acted upon by low pH. At this point, but not before, low pH can induce fusion and is in fact required for fusion to occur. However, low pH is no longer necessary after formation of the initial fusion pore: pore enlargement does not require low pH.
Assuntos
Vírus da Leucose Aviária/metabolismo , Vírus do Sarcoma Aviário/metabolismo , Membrana Celular/química , Membrana Celular/metabolismo , Fusão de Membrana , Animais , Vírus da Leucose Aviária/fisiologia , Vírus do Sarcoma Aviário/fisiologia , Fusão Celular , Linhagem Celular , Galinhas , Temperatura Baixa , Fibroblastos , Produtos do Gene env/metabolismo , Concentração de Íons de Hidrogênio , Interações Hidrofóbicas e Hidrofílicas , Lipossomos/metabolismo , Modelos Biológicos , Receptores Virais/metabolismo , Especificidade por SubstratoRESUMO
A diet high in total fat (HF) reduces hippocampal levels of brain-derived neurotrophic factor (BDNF), a crucial modulator of synaptic plasticity, and a predictor of learning efficacy. We have evaluated the capacity of voluntary exercise to interact with the effects of diet at the molecular level. Animal groups were exposed to the HF diet for 2 months with and without access to voluntary wheel running. Exercise reversed the decrease in BDNF and its downstream effectors on plasticity such as synapsin I, a molecule with a key role in the modulation of neurotransmitter release by BDNF, and the transcription factor cyclic AMP response element binding protein (CREB), important for learning and memory. Furthermore, we found that exercise influenced the activational state of synapsin as well as of CREB, by increasing the phosphorylation of these molecules. In addition, exercise prevented the deficit in spatial learning induced by the diet, tested in the Morris water maze. Furthermore, levels of reactive oxygen species increased by the effects of the diet were decreased by exercise. Results indicate that exercise interacts with the same molecular systems disrupted by the HF diet, reversing their effects on neural function. Reactive oxygen species, and BDNF in conjunction with its downstream effectors on synaptic and neuronal plasticity, are common molecular targets for the action of the diet and exercise. Results unveil a possible molecular mechanism by which lifestyle factors can interact at a molecular level, and provide information for potential therapeutic applications to decrease the risk imposed by certain lifestyles.
Assuntos
Fator Neurotrófico Derivado do Encéfalo/fisiologia , Gorduras na Dieta/efeitos adversos , Hipocampo/metabolismo , Plasticidade Neuronal/fisiologia , Condicionamento Físico Animal/fisiologia , Animais , Western Blotting , Química Encefálica/fisiologia , Proteína de Ligação ao Elemento de Resposta ao AMP Cíclico/metabolismo , Ensaio de Imunoadsorção Enzimática , Feminino , Hipocampo/efeitos dos fármacos , Aprendizagem em Labirinto/efeitos dos fármacos , Aprendizagem em Labirinto/fisiologia , Fosforilação , RNA Mensageiro/análise , Ratos , Ratos Endogâmicos F344 , Espécies Reativas de Oxigênio/análise , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Sinapsinas/metabolismoRESUMO
Infection by all enveloped viruses occurs via the fusion of viral and cellular membranes and delivery of the viral nucleocapsid into the cell cytoplasm, after association of the virus with cognate receptors at the cell surface. This process is mediated by viral fusion proteins anchored in the viral envelope and can be defined based on the requirement for low pH to trigger membrane fusion. In viruses that utilize a pH-dependent entry mechanism, such as influenza virus, viral fusion is triggered by the acidic environment of intracellular organelles after uptake of the virus from the cell surface and trafficking to a low-pH compartment. In contrast, in viruses that utilize a pH-independent entry mechanism, such as most retroviruses, membrane fusion is triggered solely by the interaction of the envelope glycoprotein with cognate receptors, often at the cell surface. However, recent work has indicated that the alpharetrovirus, avian sarcoma and leukosis virus (ASLV), utilizes a novel entry mechanism that combines aspects of both pH-independent and pH-dependent entry. In ASLV infection, the interaction of the envelope glycoprotein (Env) with cognate receptors at the cell surface causes an initial conformational change that primes (activates) Env and renders it sensitive to subsequent low-pH triggering from an intracellular compartment. Thus unlike other pH-dependent viruses, ASLV Env is only sensitive to low-pH triggering following interaction with its cognate receptor. In this manuscript we review current research on ASLV Env-receptor interactions and focus on the specific molecular requirements of both the viral fusion protein and cognate receptors for ASLV entry. In addition, we review data pertaining to the novel two-step entry mechanism of ASLV entry and propose a model by which ASLV Env elicits membrane fusion.
Assuntos
Alpharetrovirus/metabolismo , Receptores Virais/metabolismo , Proteínas do Envelope Viral/metabolismo , Alpharetrovirus/genética , Animais , Vírus da Leucose Aviária/metabolismo , Vírus do Sarcoma Aviário/metabolismo , Aves/virologia , Glicoproteínas/metabolismo , Modelos Genéticos , Receptores Virais/genéticaRESUMO
Insulin resistance and compensatory hyperinsulinaemia are thought to be the underlying factors in the metabolic or insulin-resistance syndrome and can be controlled by diet and exercise. Hyperinsulinaemia has been shown to have a direct effect on the live, suppressing the production of sex hormone-binding globulin (SHBG) and insulin-like growth factor-binding proteins 1 and 2 (IGFBP-1, -2) while stimulating the production of insulin-like growth factor 1 (IGF-1). These factors have been proposed to be important modulators of hormone-related cancers, such as prostate cancer. Men adopting a low-fat diet and daily exercise reduced their levels of serum insulin and IGF-1, while increasing their levels of IGFBP-1 and sex hormone-binding globulin (SHBG). Cell-culture studies with LNCaP prostate cancer cells showed apoptosis of tumour cells and a reduction in serum-stimulated cell growth in the post diet and exercise serum. These results suggest that prostate cancer may be another aspect of the insulin-resistance syndrome and that adopting a low-fat diet combined with regular exercise may reduce the risk for prostate and other hormone-related cancers. This needs to be tested with prospective studies.
Assuntos
Síndrome Metabólica/complicações , Neoplasias da Próstata/complicações , Humanos , MasculinoRESUMO
We have investigated a potential mechanism by which a diet, similar in composition to the typical diet of most industrialized western societies rich in saturated fat and refined sugar (HFS), can influence brain structure and function via regulation of neurotrophins. We show that animals that learn a spatial memory task faster have more brain-derived neurotrophic factor (BDNF) mRNA and protein in the hippocampus. Two months on the HFS diet were sufficient to reduce hippocampal level of BDNF and spatial learning performance. Consequent to the action of BDNF on synaptic function, downstream effectors for the action of BDNF on synaptic plasticity were reduced proportionally to BDNF levels, in the hippocampus of rats maintained on the HFS diet between 2 and 24 months. In particular, animals maintained on the HFS diet showed a decrease in levels of: (i) synapsin I mRNA and protein (total and phosphorylated), important for neurotransmitter release; (ii) cyclic AMP-response element-binding protein (CREB) mRNA and protein (total and phosphorylated); CREB is required for various forms of memory and is under regulatory control of BDNF; (iii) growth-associated protein 43 mRNA, important for neurite outgrowth, neurotransmitter release, and learning and memory. Diet-related changes were specific for the hippocampus consequent to its role in memory formation, and did not involve neurotrophin-3, another member of the neurotrophin family. Our results indicate that a popularly consumed diet can influence crucial aspects of neuronal and behavioral plasticity associated with the function of BDNF.
Assuntos
Fator Neurotrófico Derivado do Encéfalo/metabolismo , Gorduras na Dieta/efeitos adversos , Sacarose Alimentar/efeitos adversos , Hipocampo/metabolismo , Aprendizagem , Plasticidade Neuronal , Animais , Fator Neurotrófico Derivado do Encéfalo/genética , Córtex Cerebral/metabolismo , Proteína de Ligação ao Elemento de Resposta ao AMP Cíclico/metabolismo , Feminino , Proteína GAP-43/metabolismo , Hipocampo/efeitos dos fármacos , Imuno-Histoquímica , RNA Mensageiro/metabolismo , Ratos , Ratos Endogâmicos F344 , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Sinapsinas/metabolismoRESUMO
Soluble N-ethylmaleimide-sensitive fusion attachment protein receptor (SNARE)-mediated fusion of synaptic vesicles with the presynaptic-plasma membrane is essential for communication between neurons. Disassembly of the SNARE complex requires the ATPase N-ethylmaleimide-sensitive fusion protein (NSF). To determine where in the synaptic-vesicle cycle NSF functions, we have undertaken a genetic analysis of comatose (dNSF-1) in Drosophila. Characterization of 16 comatose mutations demonstrates that NSF mediates disassembly of SNARE complexes after synaptic-vesicle fusion. Hypomorphic mutations in NSF cause temperature-sensitive paralysis, whereas null mutations result in lethality. Genetic-interaction studies with para demonstrate that blocking evoked fusion delays the accumulation of assembled SNARE complexes and behavioral paralysis that normally occurs in comatose mutants, indicating NSF activity is not required in the absence of vesicle fusion. In addition, the entire vesicle pool can be depleted in shibire comatose double mutants, demonstrating that NSF activity is not required for the fusion step itself. Multiple rounds of vesicle fusion in the absence of NSF activity poisons neurotransmission by trapping SNAREs into cis-complexes. These data indicate that NSF normally dissociates and recycles SNARE proteins during the interval between exocytosis and endocytosis. In the absence of NSF activity, there are sufficient fusion-competent SNAREs to exocytose both the readily released and the reserve pool of synaptic vesicles.
Assuntos
Proteínas de Transporte/metabolismo , Proteínas de Membrana/metabolismo , Vesículas Sinápticas/metabolismo , Proteínas de Transporte Vesicular , Alelos , Animais , Proteínas de Transporte/genética , Drosophila melanogaster , Feminino , Masculino , Fusão de Membrana/fisiologia , Mutagênese , Proteínas Sensíveis a N-Etilmaleimida , Proteínas SNARE , Vesículas Sinápticas/fisiologiaRESUMO
Dietary and humoral factors are thought to be involved in the development of hypertension. This study investigated the interaction between diet and gonadal hormone status in the development and reversibility of hypertension. Normal male and female and ovariectomized (OVX) female Fischer rats were placed on either a high-fat (primarily saturated), refined carbohydrate (sucrose) (HFS) or a low-fat, complex carbohydrate (LFCC) diet at 2 mo of age, and body weight and systolic blood pressure (BP) were measured. Male and OVX female rats were initially on the diets for 7 mo, whereas normal female rats were on the diets for 2 yr. After this initial phase, a group of rats from each of the normal HFS groups were converted to the LFCC diet for a period of 1 mo (males) and 2 mo (females). The OVX females were subcutaneously implanted with a 0.5-mg estradiol (E2) pellet for 1 mo. A significant rise in arterial BP occurred within 12 mo in female and only 2 mo in male rats on the HFS diet, exceeding 140 mmHg after 24 and 7 mo, respectively. Conversion from the HFS to the LFCC diet led to a normalization of BP in both female and male rats. HFS diet-induced hypertension was accelerated by OVX in female rats, approaching the pattern seen in male rats. The effect of OVX was completely reversed by E2 replacement. BP did not significantly change in any of the LFCC groups at any time point, and E2 replacement had no effect on BP in the OVX LFCC group. All HFS groups had significantly greater body weight, with differences occurring sooner in the male and OVX rats compared with the female rats. Diet modification resulted in a partial but significant reduction of body weight, but E2 replacement did not. These results demonstrate that long-term consumption of HFS diet induces hypertension in both genders and is reversible by diet modification. Hypertension is significantly delayed in females with functional ovaries. This protection is lost by OVX and restored by estrogen replacement. Thus hormone status contributes to the delayed onset of diet-induced hypertension in females compared with males.
Assuntos
Estrogênios/farmacologia , Hipertensão/prevenção & controle , Animais , Pressão Sanguínea/efeitos dos fármacos , Peso Corporal/efeitos dos fármacos , Dieta , Estradiol/farmacologia , Terapia de Reposição de Estrogênios , Estrogênios/sangue , Feminino , Hipertensão/etiologia , Hipertensão/fisiopatologia , Masculino , Ovariectomia , Ratos , Ratos Endogâmicos F344 , Caracteres SexuaisRESUMO
High-fat feeding results in impaired insulin signaling in skeletal muscle, but the role of the insulin receptor (IR) remains controversial. In the present study, female Fischer 344 rats were fed diets either low in fat [low fat, complex carbohydrate (LFCC)] or high in fat and sucrose (HFS). Insulin-stimulated skeletal muscle glucose transport, measured in purified sarcolemmal vesicles, was lower in rats consuming the HFS diet for 2 and 8 wk compared with LFCC controls (72.9 +/- 3.5, 67.6 +/- 3.5, and 86.1 +/- 3.5 pmol x mg(-1) x 15 s(-1), respectively; P < 0.05). Muscle IR content was unchanged in 2-wk HFS animals but was 50% lower in the 8-wk HFS group (P < 0.001). However, compared with LFCC, insulin-stimulated IR autophosphorylation was 26% lower in 2-wk HFS and 40% lower in 8-wk HFS animals (P < 0.005). Total muscle content of the proposed IR inhibitors cytokine tumor necrosis factor-alpha and membrane glycoprotein PC-1 was not significantly changed in HFS animals at either 2 or 8 wk. These results demonstrate that high-fat feeding induces insulin resistance in muscle concomitant with a diminished IR signaling capacity, although the mechanism remains unknown.
Assuntos
Gorduras na Dieta/farmacologia , Resistência à Insulina/fisiologia , Receptor de Insulina/metabolismo , Animais , Transporte Biológico Ativo/efeitos dos fármacos , Western Blotting , Peso Corporal/efeitos dos fármacos , Dieta , Ensaio de Imunoadsorção Enzimática , Feminino , Glucose/metabolismo , Transportador de Glucose Tipo 1 , Técnicas In Vitro , Insulina/farmacologia , Resistência à Insulina/genética , Membranas/metabolismo , Proteínas de Transporte de Monossacarídeos/metabolismo , Músculo Esquelético/efeitos dos fármacos , Músculo Esquelético/metabolismo , Fosforilação , Ratos , Ratos Endogâmicos F344 , Receptor de Insulina/efeitos dos fármacos , Transdução de Sinais/efeitos dos fármacos , Sacarose/farmacologia , Fator de Necrose Tumoral alfa/metabolismoRESUMO
PURPOSE: A high fat diet and sedentary lifestyle may predispose men to prostate cancer through effects on serum factors such as hormones. We evaluated the effects of a low fat, high fiber diet and exercise intervention on serum stimulated growth of established prostate cancer cell lines. MATERIALS AND METHODS: Fasting serum was obtained from 13 overweight men before and after undergoing an 11-day low fat, high fiber diet and exercise intervention. Serum was also obtained from 8 men who had complied with the regimen for a mean of 14.2 years. Hormone dependent LNCaP and independent PC-3 prostate cancer cell lines were grown in culture medium containing 10% of subject pre-intervention or post-intervention serum and viable cells were counted after 48 hours. Anthropometry, serum free testosterone, lipids and glucose were measured in all subjects. RESULTS: Post-intervention serum from each of the 11-day intervention subjects reduced LNCaP cell growth by a mean of 30% compared with pre-intervention serum from each (p <0.01). LNCaP cell growth in serum from long-term subjects was 15% below that of post-intervention serum (p <0.01). There was no difference in the growth of PC-3 cells when cultured with serum from either intervention group. Serum free testosterone, body weight, glucose and lipids were significantly reduced in 11-day subjects. CONCLUSIONS: A low fat, high fiber diet and exercise intervention resulted in serum changes that significantly reduced the growth of androgen responsive LNCaP prostate cancer cells in vitro.
Assuntos
Dieta , Exercício Físico , Neoplasias da Próstata/patologia , Neoplasias da Próstata/prevenção & controle , Adulto , Idoso , Divisão Celular , Gorduras na Dieta/administração & dosagem , Fibras na Dieta/administração & dosagem , Humanos , Masculino , Pessoa de Meia-Idade , Células Tumorais CultivadasRESUMO
This study was designed to examine whether abnormalities that comprise the metabolic syndrome, including insulin resistance, hyperinsulinemia, hypertension, hyperlipidemia, and obesity, are reversible by diet. Female Fischer rats were placed on either a high-fat, refined-carbohydrate (HFS) diet or low-fat, complex-carbohydrate (LFCC) diet for a period of 20 months. After 20 months, a group of HFS rats were switched to the LFCC diet (HFS/LFCC) for a period of 2 months. Skeletal muscle glucose transport, plasma insulin, systolic blood pressure, and plasma lipids were measured in all groups after 22 months. Energy intake and body weight were measured weekly. In the HFS group, insulin-stimulated glucose transport was significantly reduced (67+/-4 versus 98+/-4 pmol. mg(-)(1). 15 s(-)(1)), whereas plasma insulin (300+/-49 versus 82+/-8 pmol/L), blood pressure (147+/-4 versus 123+/-4 mm Hg), plasma triglycerides (2.58+/-0.31 versus 0.39+/-0.04 mmol/L), LDL cholesterol (C) (3.45+/-0.40 versus 0.89+/-0.06 mmol/L), LDL-C to HDL-C ratio (2.9+/-0.1 versus 2.2+/-0.1), VLDL-C (1.53+/-0.23 versus 0.37+/-0.07 mmol/l), Total-C (5.56+/-0.58 versus 1.49+/-0.10 mmol/L), and body weight (360+/-11 versus 260+/-5 g) were all significantly elevated compared with the LFCC. Energy intake did not differ significantly; however, the LFCC had a much poorer feed efficiency. Conversion to a LFCC diet for 2 months led to normalization of glucose transport, blood pressure, plasma insulin, and VLDL-C and significant amelioration of obesity and other lipid abnormalities. These results demonstrate that syndrome X induced by an inappropriate diet is reversed with implementation of a low-fat, unrefined-carbohydrate diet without caloric restriction and suggest that diet may be a possible treatment for multiple simultaneous cardiovascular risk factors.
Assuntos
Comportamento Alimentar , Angina Microvascular/dietoterapia , Animais , Transporte Biológico , Pressão Sanguínea , Peso Corporal , Doença Crônica , Dieta com Restrição de Gorduras , Modelos Animais de Doenças , Ingestão de Energia , Feminino , Glucose/metabolismo , Insulina/sangue , Lipídeos/sangue , Angina Microvascular/sangue , Angina Microvascular/fisiopatologia , Ratos , Ratos Endogâmicos F344RESUMO
We have recently demonstrated that long-term consumption of a high-fat, refined-carbohydrate (HFS) diet induces hypertension (HTN) in normal rats compared with a low-fat, complex-carbohydrate (LFCC) diet. Limited evidence suggests that high-fat or high-sugar diets cause enhanced generation of reactive oxygen species (ROS). We therefore hypothesized that by inducing oxidative stress, the HFS diet may promote nitric oxide (NO) inactivation and HTN. To test this hypothesis, female Fischer rats were placed on either the HFS or the LFCC diet starting at 2 months of age. Blood pressure, urinary NO metabolites (NO(x)), and total renal NO synthase activity were monitored, and the tissue abundance of nitrotyrosine (NT), which is the stable "footprint" of NO oxidation by ROS, was determined. The HFS diet group exhibited a gradual rise in arterial blood pressure and were hypertensive by 18 months. This trend was accompanied by a marked accumulation of NT in all tested tissues, an initial rise and a subsequent fall in NO synthase activity, and a fall in urinary NO(x) excretion. The HFS diet-fed animals had a blunted blood pressure response to the NO synthase inhibitor N:(omega)-nitro-L-arginine methyl ester (L-NAME) compared with the LFCC diet group, which showed a marked hypertensive response to L-NAME. L-NAME-induced HTN was reversible with L-arginine in the LFCC diet group; however, HTN was not corrected by L-arginine supplementation in the HFS diet group. These findings point to enhanced ROS-mediated inactivation and sequestration of NO, which may contribute to the reduction of bioactive NO and HTN in the HFS diet-fed animals.
Assuntos
Carboidratos da Dieta/administração & dosagem , Gorduras na Dieta/administração & dosagem , Hipertensão/etiologia , Óxido Nítrico/metabolismo , Espécies Reativas de Oxigênio/metabolismo , Tirosina/análogos & derivados , Tirosina/metabolismo , Animais , Arginina/farmacologia , Biomarcadores , Pressão Sanguínea , Peso Corporal , Inibidores Enzimáticos/farmacologia , Feminino , NG-Nitroarginina Metil Éster/farmacologia , Óxido Nítrico/urina , Óxido Nítrico Sintase/antagonistas & inibidores , Óxido Nítrico Sintase/metabolismo , Oxirredução , Estresse Oxidativo/fisiologia , Ratos , Ratos Endogâmicos F344RESUMO
Hyperhomocysteinemia (HH) and hyperinsulinemia are both risk factors for cardiovascular disease. To examine the effects of hyperinsulinemia on homocysteine metabolism, we fed rats a high-fat-sucrose (HFS) diet and then measured the hepatic mRNA and activity of 2 key enzymes involved in this metabolic pathway: 5,10-methylenetetrahydrofolate reductase (MTHFR) and cystathionine-beta-synthase (CbetaS). Fischer rats made insulin-resistant by a HFS diet were examined at 6 months and 2 years of age and compared with control rats fed a low-fat, complex-carbohydrate (LFCC) diet. At the end of 6 months, the HFS rats were heavier than the LFCC rats (214 +/- 3.4 v 188 +/- 1.4 g, P < .01). There were no differences in blood glucose between HFS and LFCC rats; however, plasma insulin and homocysteine concentrations were elevated in HFS rats (insulin, 56 +/- 12 v 14.5 +/- 2.9 microU/mL; homocysteine, 10.77 +/- 0.9 v 6.89 +/- 0.34 micromol/L, P < .01). Hepatic CbetaS enzyme activity was significantly lower in HFS compared with LFCC rats (0.45 v 0.64 U/mg, P = .0001), and this decrease was reflected in a decrease of the CbetaS mRNA concentration. In contrast, hepatic MTHFR enzyme activity and mRNA concentration were significantly elevated in the HFS group compared with controls (HFS and LFCC, 8.62 and 4.8 nmol/h/mg protein, respectively, P = .0001). These changes in plasma homocysteine, CbetaS, and MTHFR were significantly correlated with the degree of obesity and hyperinsulinemia. Fasting plasma insulin correlated significantly and positively with plasma homocysteine (r = .51, P < .01) and MTHFR activity (r = .48, P < .01) and negatively with CbetaS activity (r = -.54, P < .001). CbetaS and MTHFR activities were inversely correlated with each other (r = -.58, P < .001). In conclusion, rats fed a HFS diet are hyperinsulinemic, and the hyperinsulinemia is associated with an elevated homocysteine concentration and changes in 2 key enzymes in homocysteine metabolism.
Assuntos
Dieta , Gorduras na Dieta/farmacologia , Sacarose Alimentar/farmacologia , Homocisteína/sangue , Hiperinsulinismo/enzimologia , Fígado/enzimologia , Fatores Etários , Animais , Peso Corporal , Cistationina beta-Sintase/genética , Cistationina beta-Sintase/metabolismo , Feminino , Hiperinsulinismo/sangue , Hiperinsulinismo/etiologia , Insulina/sangue , Metilenotetra-Hidrofolato Redutase (NADPH2) , Oxirredutases atuantes sobre Doadores de Grupo CH-NH/genética , Oxirredutases atuantes sobre Doadores de Grupo CH-NH/metabolismo , RNA Mensageiro/metabolismo , Ratos , Ratos Endogâmicos F344 , Análise de RegressãoRESUMO
Dietary factors including fat and fiber have been reported to play a role in the development of breast cancer, possibly mediated by changes in estradiol. Diet and exercise have been shown to affect levels of sex hormone-binding globulin (SHBG), which in turn regulate the bioavailability of estradiol. Diet and exercise also affect insulin levels, which play a role in the synthesis of SHBG, and the hormone itself is a potent mitogen for many cancer cell lines. This study was designed to measure the effects of a low-fat, high-fiber diet, combined with regular aerobic exercise, on the levels of SHBG, insulin, and serum lipids in postmenopausal women with or without hormone replacement therapy (HRT). Two groups of postmenopausal women, 11 on HRT and 11 not on HRT, underwent a low-fat (1O% fat calories), high-fiber (65-70 g/day) diet-and-exercise intervention for three weeks. Serum SHBG, insulin, and lipids were measured before and after the regimen. After the intervention, SHBG levels were significantly increased for the women on HRT (44.5 +/- 3.4 vs. 62 +/- 6.4 nmol/l) and the women not on HRT (32.1 +/- 4.6 vs. 45.5 +/- 6.1 nmol/l, both changes p < 0.01). Also after the intervention, insulin levels were significantly reduced for the women on HRT (196 +/- 44.4 vs. 119.8 +/- 28.7 pmol/l) and the women not on HRT (144.2 +/- 17.9 vs. 115.5 +/- 20.8 pmol/l, both changes p < 0.01). Body mass index and total cholesterol were significantly reduced for both groups of women (all changes p < 0.01). Although the exact mechanism for the change in SHBG is not known, the increases in SHBG and reductions in insulin as a result of this lifestyle intervention should reduce the risk for breast cancer in postmenopausal women.
Assuntos
Dieta com Restrição de Gorduras , Fibras na Dieta/administração & dosagem , Exercício Físico/fisiologia , Insulina/sangue , Lipídeos/sangue , Globulina de Ligação a Hormônio Sexual/metabolismo , Antropometria , Neoplasias da Mama/sangue , Neoplasias da Mama/prevenção & controle , Gorduras na Dieta/administração & dosagem , Estradiol/sangue , Feminino , Terapia de Reposição Hormonal , Humanos , Pessoa de Meia-Idade , Pós-MenopausaRESUMO
This study examined the effects of acute exercise on skeletal muscle nitric oxide synthase (NOS) activity. Female Sprague-Dawley rats were divided into three groups: control, exercise, and exercise + N(G)-nitro-L-arginine methyl ester (L-NAME). In the exercise + L-NAME group, L-NAME was administered in the drinking water (1 mg/ml) for 2 days and subsequently the exercise and exercise + L-NAME groups underwent a 45-min bout of exhaustive treadmill running after which NOS activity and muscle glycogen were measured. In the control and exercise groups, 1-amino-S-methylisothiourea (AMITU), a selective neuronal NOS inhibitor, with and without additional nonselective NOS blockade [with N(G)-monomethyl-L-arginine (L-NMMA)], was used in vitro to assess the contribution of nNOS to total NOS activity. The exercise bout increased NOS activity by 37% in exercise compared with control groups, and both groups had significantly greater NOS activity compared with exercise + L-NAME. AMITU decreased total NOS activity in the control and exercise groups by 31.8 and 30.2%, respectively, and these activities were significantly greater than AMITU + L-NMMA in both control and exercise groups. We conclude that 1) there is basal neuronal NOS and endothelial NOS activity in skeletal muscle, 2) an acute exercise bout increases NOS activity in skeletal muscle, and 3) glycogen depletion during exercise occurs irrespective of NOS activity.
Assuntos
Músculo Esquelético/enzimologia , Óxido Nítrico Sintase/metabolismo , Condicionamento Físico Animal/fisiologia , Animais , Feminino , Glicogênio/metabolismo , Atividade Motora/fisiologia , Músculo Esquelético/metabolismo , Ratos , Ratos Sprague-Dawley , Fatores de TempoRESUMO
This study was designed to examine the relationship between diet-induced insulin resistance/hyperinsulinemia, fat cell hypertrophy, and hormone-sensitive lipase (HSL) to elucidate whether an attenuated HSL activity leads to obesity. Female Fischer 344 rats were fed either a low-fat, complex-carbohydrate diet or a high-fat, refined-sugar (HFS) diet for 2 wk, 2 mo, or 6 mo. Adipose tissue morphology and HSL activity as well as plasma free fatty acid and glycerol levels were determined at these times. No differences between groups were seen after 2 wk except the previously reported hyperinsulinemia in the HFS animals. At both 2 and 6 mo, the HFS animals demonstrated adipocyte hypertrophy. Basal and stimulated HSL activities and plasma glycerol were significantly elevated in the HFS group. There was a positive correlation between adipocyte size and HSL activity for both basal and stimulated states. These results demonstrate that an attenuated HSL activity is not observed with the onset of insulin resistance/hyperinsulinemia and therefore does not play a role in the development of obesity.
Assuntos
Adipócitos/citologia , Adipócitos/enzimologia , Dieta , Esterol Esterase/metabolismo , Adipócitos/efeitos dos fármacos , Animais , Tamanho Celular , Carboidratos da Dieta/administração & dosagem , Gorduras na Dieta/administração & dosagem , Sacarose Alimentar/administração & dosagem , Ácidos Graxos não Esterificados/sangue , Feminino , Glicerol/sangue , Humanos , Insulina/sangue , Resistência à Insulina , Lipólise/efeitos dos fármacos , Obesidade/enzimologia , Obesidade/etiologia , Propranolol/farmacologia , Ratos , Ratos Endogâmicos F344RESUMO
PURPOSE: Hypertension (HTN), hyperlipidemia (HLP), and hyperinsulinemia are known risk factors for the development of cardiovascular disease. Each has independently been shown to be associated with impaired endothelial function, as demonstrated by decreased endothelial derived relaxation (EDR). Previous work in our laboratory has shown that rats fed a high-fat sucrose (HFS) diet will become insulin resistant, hypertriglyceridemic, and hypertensive. We hypothesize that the development of these diet-induced risk factors is associated with endothelial dysfunction and a significant decrease in EDR. Furthermore, the endothelial dysfunction will be improved by returning to a normal (low-fat complex carbohydrate (LFCC)) diet. METHODS: Adult, male Fischer rats were fed either a LFCC or a HFS diet for 6 months (n = 8 in each group). A third group of rats (SWITCH) was fed a HFS diet for 6 months and then changed to a LFCC diet for 4 weeks. Blood pressure was measured via the tail-cuff method weekly. The rats were sacrificed and aortic ring segments were placed in physiologic tissue baths for measurement of vascular reactivity to various agents. Arterial ring segments were constricted with potassium chloride (K) and phenylephrine (PE). Endothelial-dependent vasorelaxation was measured with acetylcholine (Ach), bradykinin (BK), and calcium ionophore (CA). Endothelial-independent relaxation was measured using sodium nitroprusside (NTP). RESULTS: The HFS diet group developed HTN compared to LFCC group. Vasoconstriction to K and PE were similar in all groups. Vasorelaxation to Ach, BK, and CA was significantly decreased in the HFS group, but returned to baseline in the diet-switched group, as did the systolic blood pressure. There were no differences in relaxation to NTP. CONCLUSIONS: HFS diet-induced HTN is associated with significantly decreased EDR. Switching to a low-fat diet reverses this effect. The vascular smooth muscle contraction and endothelial-independent relaxation are not affected by the diet-induced risk factors. There is a direct and reversible effect of an HFS diet on endothelial function and blood pressure.