Perda de glicosaminoglicanas da cartilagem após ruptura do ligamento cruzado anterior: influência do tempo de ruptura e da lesão condral

OBJECTIVE: To quantify the concentration of sulfated glycosaminoglycans (GAGs) concentration in the synovial fluid (SF) of knees with chronic anterior cruciate ligament (ACL) rupture and to identify possible associations between GAG concentration in SF and the time elapsed since rupture and degree of chondral injury. METHOD: Fourteen adult male subjects with total unilateral ACL rupture, which had occurred between 5 and 144 months earlier, were assessed. All subjects underwent joint aspiration; it was possible to collect SF from ten individuals. The samples were quantified to determine the GAG concentration using dimethylmethylene blue (DMMB) staining. The degree of chondral injury was macroscopically evaluated using the modified Mankin histological scale. Spearman correlation test (< 0.05) was used to evaluate the association between GAG concentration and chondral injury, and Pearson correlation test (< 0.05) was used to evaluate the association between GAG concentration and the time elapsed since rupture. RESULTS: The GAG concentration in SF showed a mean variation of 73.84 &plusmn; 40.75 µg/ml, with a mean time of 40.4 &plusmn; 40.3 months since the rupture. There was no correlation between GAG concentration and time since the rupture (r= -0.09, p= 0.81). The chondral injury grades found were 0, 1, 4 and 5. There was no correlation between chondral injury grade and GAG concentration in SF (r= -0.41, p= 0.24). CONCLUSION: After at least 5 months, the GAG concentration in SF from knees with ACL rupture is independent of the time elapsed since rupture and/or the severity of chondral injury.OBJETIVO: Quantificar a concentração de glicosaminoglicanas sulfatadas (GAGs) no líquido sinovial (LS) de joelhos com ruptura crônica do ligamento cruzado anterior (LCA) e identificar uma possível correlação entre a concentração de GAGs no LS e o tempo pós-ruptura e grau de lesão condral. MÉTODOS: Foram avaliados 14 indivíduos adultos do sexo masculino com ruptura total unilateral do LCA, ocorrida entre cinco a 144 meses. Todos os sujeitos foram puncionados, sendo possível a coleta de LS em dez indivíduos. As amostras foram quantificadas para determinar a concentração de GAGs usando a coloração azul de dimetilmetileno, método descrito por Farndale21. O grau de lesão condral foi macroscopicamente avaliado pela escala histológica de Mankin modificada por Messner14. As correlações entre concentração de GAGs e lesão condral foram feitas pelo teste de correlação de Sperman (p< 0,05) e a concentração de GAGs e tempo pós-ruptura pelo teste de correlação de Pearson (p< 0,05). RESULTADOS: Concentração de GAGs no LS apresentou variação média de 73,84 &plusmn; 40,75µg/mL, sendo o tempo médio pós-ruptura de 40,4 + 40,3 meses. Não houve correlação entre concentração de GAGs e o tempo pós-ruptura (r= -0,09, p= 0,81). Os graus de lesão condral encontrados foram de 0, 1, 4 e 5. Não houve correlação entre grau de lesão condral e a concentração de GAGs no LS (r= -0,41, p= 0,24). CONCLUSÕES: Após no mínimo cinco meses, a concentração de GAGs no LS de joelhos com ruptura do LCA independe do tempo pós-ruptura e/ou do grau de lesão condral.646

Physical Exercise Decreases Fasting Hyperglycemia in Diabetic Mice Through AMPK Activation

Introduction: The deficiency in glucose uptake in peripheral tissues and increased hepatic gluconeogenesis are physiopathological phenomena observed in type 2 diabetes patients. Physical exercise plays an important role in the improvement of glycemic profile in diabetic patients; however, the mechanisms involved in these processes have not been fully elucidated. Objective: to assess the role of AMPK protein in the glycemic control of diabetic mice after exercise. Methods: During fasting condition, the insulin tolerance test (ITT) and Western blot technique, were combined to assess the glucose homeostasis in diabetic mice (ob/ob and db/db) after a single swimming session. Results: Fasting hyperglycemia, severe insulin resistance and deficiency in the AMPk/ACC signaling in muscle and liver observed in the diabetic mice were reversed after the exercise session. The restoration of AMPK/ACC signaling reduced the expression of the gluconeogenic enzyme, PEPCk in the liver, and increased the translocation of GLUT4 in the skeletal muscle. These data indicate that the activation of AMPK/ACC pathway induced by physical exercise is important to reduce fasting glucose levels in experimental models of type 2 diabetes. These data open new insights for determination of physical activity control on the glucose homeostasis in diabetic patients.15317918

IL-6 and IL-10 Anti-Inflammatory Activity Links Exercise to Hypothalamic Insulin and Leptin Sensitivity through IKK beta and ER Stress Inhibition

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)Overnutrition caused by overeating is associated with insulin and leptin resistance through IKK beta activation and endoplasmic reticulum (ER) stress in the hypothalamus. Here we show that physical exercise suppresses hyperphagia and associated hypothalamic IKK beta/NF-k beta activation by a mechanism dependent upon the pro-inflammatory cytokine interleukin (IL)-6. The disruption of hypothalamic-specific IL-6 action blocked the beneficial effects of exercise on the re-balance of food intake and insulin and leptin resistance. This molecular mechanism, mediated by physical activity, involves the anti-inflammatory protein IL-10, a core inhibitor of IKK beta/NF-k beta signaling and ER stress. We report that exercise and recombinant IL-6 requires IL-10 expression to suppress hyperphagia-related obesity. Moreover, in contrast to control mice, exercise failed to reverse the pharmacological activation of IKK beta and ER stress in C3H/HeJ mice deficient in hypothalamic IL-6 and IL-10 signaling. Hence, inflammatory signaling in the hypothalamus links beneficial physiological effects of exercise to the central action of insulin and leptin.88Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq

Central Exercise Action Increases the AMPK and mTOR Response to Leptin

AMP-activated protein kinase (AMPK) and mammalian Target of Rapamycin (mTOR) are key regulators of cellular energy balance and of the effects of leptin on food intake. Acute exercise is associated with increased sensitivity to the effects of leptin on food intake in an IL-6-dependent manner. To determine whether exercise ameliorates the AMPK and mTOR response to leptin in the hypothalamus in an IL-6-dependent manner, rats performed two 3-h exercise bouts, separated by one 45-min rest period. Intracerebroventricular IL-6 infusion reduced food intake and pretreatment with AMPK activators and mTOR inhibitor prevented IL-6-induced anorexia. Activators of AMPK and fasting increased food intake in control rats to a greater extent than that observed in exercised ones, whereas inhibitor of AMPK had the opposite effect. Furthermore, the reduction of AMPK and ACC phosphorylation and increase in phosphorylation of proteins involved in mTOR signal transduction, observed in the hypothalamus after leptin infusion, were more pronounced in both lean and diet-induced obesity rats after acute exercise. Treatment with leptin reduced food intake in exercised rats that were pretreated with vehicle, although no increase in responsiveness to leptin-induced anorexia after pretreatment with anti-IL6 antibody, AICAR or Rapamycin was detected. Thus, the effects of leptin on the AMPK/mTOR pathway, potentiated by acute exercise, may contribute to appetite suppressive actions in the hypothalamus