الملخص
SUMMARY: Nonalcoholic fatty liver disease (NAFLD) might progress the steatosis to nonalcoholic steatohepatitis (NASH), reaching a cirrhosis state and possibly hepatocellular carcinoma. The liver of three-month-old C57BL/6J mice (wild-type, WT group, n=10) and leptin- deficient obese mice (ob/ob group, n=10) were studied, focusing on the mechanisms associated with the activation of the hepatic stellate cells (HSCs) and pro-fibrogenesis. The obese ob/ob animals' liver showed steatosis, increased lipogenesis gene expressions, inflammation, increased pro-inflammatory gene expressions, inflammatory infiltrate, and potential apoptosis linked to a high Caspase 3 expression. In ob/ob mice, liver sections were labeled in the fibrotic zones by anti-alpha-smooth muscle actin (alpha-SMA) and anti-Reelin, but not in the WT mice. Moreover, the alpha-SMA gene expression was higher in the ob/ob group's liver than the WT group. The pro-fibrogenic gene expressions were parallel to anti- alpha-SMA and anti-Reelin immunofluorescence, suggesting HSCs activation. In the ob/ob animals, there were increased gene expressions involved with lipogenesis (Peroxisome proliferator-activated receptor-gamma, Cell death-inducing DFFA-like effector-c, Sterol regulatory element-binding protein-1c, and Fatty acid synthase), pro-fibrogenesis (Transforming growth factor beta1, Smad proteins- 3, Yes-associated protein-1, Protein platelet-derived growth factor receptor beta), pro-inflammation (Tumor necrosis factor-alpha, and Interleukin-6), and apoptosis (Caspase 3). In conclusion, the results in obese ob/ob animals provide a clue to the events in humans. In a translational view, controlling these targets can help mitigate the hepatic effects of human obesity and NAFLD progression to NASH.
RESUMEN: La enfermedad del hígado graso no alcohólico (HGNA) puede progresar de la esteatosis a esteatohepatitis no alcohólica (ENA), alcanzando un estado de cirrosis y posiblemente carcinoma hepatocelular. Se estudió el hígado de ratones C57BL / 6J de tres meses de edad (tipo salvaje, grupo WT, n = 10) y ratones obesos con deficiencia de leptina (grupo ob/ob, n = 10), centrándose en los mecanismos asociados con la activación de las células estrelladas hepáticas (HSC) y profibrogénesis. El hígado de los animales obesos ob/ob mostró esteatosis, aumento de la expresión génica de la lipogénesis, inflamación, aumento de la expresión génica proinflamatoria, infiltrado inflamatorio y posible apoptosis ligada a una alta expresión de Caspasa 3. En ratones ob/ob, las sec- ciones de hígado se marcaron en las zonas fibróticas con anti-alfa- actina de músculo liso (alfa-SMA) y anti-Reelin, pero no en los ratones WT. Además, la expresión del gen alfa-SMA fue mayor en el hígado del grupo ob/ob que en el grupo WT. Las expresiones génicas profibrogénicas fueron paralelas a la inmunofluorescencia anti-alfa-SMA y anti-Reelin, lo que sugiere la activación de las HSC. En los animales ob/ob, hubo un aumento de las expresiones génicas involucradas con la lipogénesis (receptor activado por proliferador de peroxisoma gamma, efector c similar a DFFA inductor de muerte celular, proteína de unión al elemento regulador de esterol-1c y sintasa de ácidos grasos), pro-fibrogénesis (factor de crecimiento transformante beta 1, proteínas Smad-3, proteína-1 asociada a Yes, receptor beta del factor de crecimiento derivado de plaquetas de proteínas), proinflamación (factor de necrosis tumoral alfa e interleucina-6) y apoptosis (caspasa 3). ). En conclusión, los resultados en animales obesos ob/ob proporcionan una pista de los eventos en humanos. Desde un punto de vista traslacional, el control de estos objetivos puede ayudar a mitigar los efectos hepáticos de la obesidad humana y la progresión de HGNA a ENA.
الموضوعات
Animals , Mice , Leptin/deficiency , Fatty Liver/pathology , Photomicrography , Apoptosis , Microscopy, Confocal , Lipogenesis/genetics , Caspase 3/metabolism , Hepatic Stellate Cells/ultrastructure , Fatty Liver/genetics , Real-Time Polymerase Chain Reaction , Non-alcoholic Fatty Liver Disease/pathology , Inflammation/genetics , Liver/ultrastructure , Liver Cirrhosis/genetics , Liver Cirrhosis/pathology , Mice, Inbred C57BL , Obesityالملخص
O treinamento físico aeróbio (TFA) tem sido cada vez mais recomendado para a prevenção e tratamento de comorbidades, que podem ter em comum a resistência periférica à insulina (RI), como o diabetes tipo 2 (DM2) ea obesidade. Caracterizada pelo acúmulo excessivo de tecido adiposo, a obesidade também implica a disfunção endócrina desse tecido. Leptina e adiponectina são hormônios secretados pelos adipócitos que desempenham importante função no metabolismo energético, cujo desequilíbrio está fortemente relacionado à obesidade e à RI. Fisiologicamente, mulheres apresentam maiores níveis de leptina e adiponectina comparados aos de homens. Contudo, em condições como a obesidade e/ou DM2 esses níveis diminuem, deixando o organismo de fêmeas sem os efeitos protetores dessas adipocinas. Assim, o presente estudo teve como objetivo investigar os efeitos do TFA sobre as características morfofuncionais de TAB e pâncreas de camundongos fêmeas com deficiência de leptina. Para isso, camundongos fêmeas com oitosemanas de idade C57BL/J6 selvagem (C57) ou com deficiência de leptina ob/ob (Lepob) foram separadas nos grupos sedentárias (S) e treinadas (T): C57S (n=6), C57T (n=6), LepobS (n=5) e LepobT (n=8). O TFA foi realizadodurante oito semanas, com corrida a 60% da capacidade máxima, 1h/dia, 5x/semana. Água e ração foram administradas ad libitum. Os procedimentos foram aprovados pelo CEUA da EACH-USP (#001/2017). O acompanhamento do peso corporal e consumo de ração foi realizado semanalmente. Na 7ª semana de protocolo foram realizados os testesde tolerância àglicose (TTG) e à insulina (TTI). Ao final do protocolode treinamento foram avaliados parâmetros metabólicos de repouso e de esforço através da calorimetria indireta. A eutanásia dos animais foi realizada cominjeção intraperitoneal de anestésico e, em seguida, foram coletados os tecidos adiposos branco (TAB) subcutâneo e retroperitoneal e a porção esplênica do pâncreas, utilizados, posteriormente, para análises morfológicas. A determinação de adiponectinasérica foi realizada através de kit ELISA. Os resultados foram analisados por ANOVA de duas vias mais o teste post-hoc de Bonferroni. O protocolo de TFA não foi eficaz em reduzir a adiposidade e hiperfagia dos animais LepobS e LepobT, contudo o grupo LepobT apresentou tamanho reduzido de adipócitos doTAB-SC comparado ao grupo LepobS.O TFA não afetouo metabolismo glicêmico dos animais. Os parâmetros metabólicos derepouso não apresentaram alteração, contudo, noteste de esforço físico máximo, o grupo LepobT apresentou maiores tempo em teste até a exaustão e iVO2 comparado ao grupo LepobS. Por fim, embora o TFA não tenha aumentadoa secreção de adiponectina, o que poderia regular positivamente o metabolismo glicêmico de camundongos deficientes em leptina, melhorou a condição metabólica desses animais e reduziu a hipertrofia dos adipócitos do TAB-SC
Aerobic physical training (APT) has become an important strategy for the prevention and treatment of comorbidities, which may have in common the peripheral resistance to insulin(RI), such as type 2 diabetes and obesity. Characterized by the excessive accumulation of adipose tissue, obesity also implies endocrine dysfunction of this tissue.Leptin and adiponectin are hormones secreted by adipocytes that play an important role in energy metabolism, whose imbalance is strongly related to obesity and RI. Physiologically, women have higher levels of leptin and adiponectin compared to men.However,in conditions such as obesity and/or type 2 diabetes these levels decrease, leaving the body of females without the protective effects of these adipokines.Thus, the present study aimed to investigate the effects of APTon the morphofunctional characteristics of white adipose tissue (WAT)and pancreas in leptin-deficientfemale mice. For this, 8-week-old wild C57BL/J6 (C57) or leptin-deficient ob/ob (Lepob) female mice were separated into the groups sedentary (S) and trained (T): C57S (n = 6), C57T (n = 6), LepobS (n = 5) and LepobT (n = 8). Mice were housed in a temperature-controlled (22 ± 2°C) and 12-h light/12-h dark cycle, with free access to tap water and food ad libitum. The APT was performed for 1 h/day at 60% of maximal velocity achieved in the running capacity test, five times per week for eight weeks. The procedures were performed in accordance with the guidelines of the Brazilian College for Animal Experimentation and were approved by the Ethics Committee of the School of Arts, Sciences and Humanities of University of Sao Paulo (Process number001/2017). Body weight and feed intake were evaluated weekly. In the 7th week of protocol, the glucose and insulin tolerance tests (GTT and ITT) were performed. At the end of the training protocol, metabolic parameters of resting and maximal exertion were evaluated through indirect calorimetry. The euthanasia of the animals was performed by intraperitoneal injection of anaesthetic and thenthe subcutaneous and retroperitoneal WAT and the splenic portion of the pancreas were collected, and subsequentlyused for morphological analysis. Serum adiponectin quantification was performed by ELISA kit. The results were analyzed by two-way ANOVA plus the Bonferroni post-hoc test. The APT protocol was not effective in reducing the adiposity and hyperphagia of the animals LepobS and LepobT, however the LepobT group presented a reduced size of WAT-SC adipocytes compared to the LepobS group. Also, APT did not affectthe glycemic metabolism of theseanimals. Although the resting metabolic parameters were not different, in the maximal physical exercise test, the LepobT group had longer time to exhaustion test and iVO2 compared to the LepobS group.Finally, although APTdid not increase adiponectin plasma concentration, which could positively regulate the glycemic metabolism of leptin-deficient mice, it improved the metabolic condition of these animals and reduced the adipocyte hypertrophy of WAT-SC
الموضوعات
Animals , Female , Mice , Exercise/physiology , Leptin/deficiency , Insulin Resistance , Diabetes Mellitus, Type 2/prevention & control , Obesity/prevention & controlالملخص
The clinical manifestations of lipodystrophy syndromes (LS) are hypoleptinemia, hyperglycemia, insulin resistance, dyslipidemia and hepatic steatosis. Leptin replacement therapy (LRT) is effective at improving these pathologies. Currently, there are no data compiling the evidence from the literature, and demonstrating the effect of LRT in LS patients. A systematic review of the MEDLINE and Cochrane Library databases was conducted to identify studies assessing the effect of LRT on metabolic and hepatic endpoints in patients with LS not associated with highly active antiretroviral therapy (HAART) use. Standardized mean differences (SMD) and 95% confidence intervals of pooled results were calculated for overall changes in glucose homeostasis, lipid profile, and hepatic physiology, using an inverse-variance random-effects model. After screening, 12 studies were included for review. Meta-analysis of results from 226 patients showed that LRT decreased fasting glucose [0.75 SMD units (range 0.36‐1.13), p=0.0001], HbA1c [0.49 (0.17‐0.81), p=0.003], triglycerides [1.00 (0.69‐1.31), p<0.00001], total cholesterol [0.62 (0.21‐1.02), p=0.003], liver volume [1.06 (0.51‐1.61), p=0.0002] and AST [0.41 (0.10‐0.73) p=0.01]. In patients with non-HAART LS, LRT improves the outcome of several metabolic and hepatic parameters. Studies were limited by small populations and therefore large prospective trials are needed to validate these findings.
As manifestações clínicas das síndromes lipodistróficas (SL) incluem hipoleptinemia, hiperglicemia, resistência insulínica, dislipidemia e esteatose hepática. A terapia de reposição de leptina (TRL) melhora tais parâmetros, mas atualmente não há dados compilados demonstrando tal efeito. Uma revisão sistemática dos bancos de dados MEDLINE e Cochrane Library identificou estudos avaliando os efeitos da TRL sobre parâmetros metabólicos e hepáticos em pacientes com SL não associadas ao uso de antirretrovirais. Diferenças médias padronizadas (DMP) e intervalos de confiança de 95% foram calculados a partir dos resultados, para os efeitos da TRL sobre a homeostase da glicose, perfil lipídico, e morfologia/função hepática, usando um modelo de variação inversa e efeitos randômicos. Após a triagem, 12 estudos foram incluídos para revisão. A metanálise dos resultados de 226 pacientes mostrou que a TRL reduziu a glicemia de jejum [0,75 DMP (amplitude 0,36‐1,13), p=0,0001], HbA1c [0,49 (0,17‐0,81), p=0,003], triglicerídeos [1,00 (0,69‐1,31), p<0,00001], colesterol total [0,62 (0,21‐1,02), p=0,003], volume hepático [1,06 (0,51‐1,61), p=0,0002] e AST [0,41 (0,10‐0,73), p=0,001]. Em pacientes com SL não associada ao uso de antirretrovirais, a TRL melhora vários parâmetros metabólicos e hepáticos. Os estudos avaliados foram limitados pelo pequeno número de pacientes. Maiores estudos clínicos prospectivos são necessários para validar tais achados.
الموضوعات
Humans , Hormone Replacement Therapy , Leptin/therapeutic use , Lipodystrophy/drug therapy , Antiretroviral Therapy, Highly Active , Blood Glucose/metabolism , Cholesterol/metabolism , Fatty Liver/drug therapy , Fatty Liver/metabolism , Glycated Hemoglobin/metabolism , Insulin/metabolism , Leptin/deficiency , Leptin/metabolism , Lipodystrophy/metabolism , Liver/metabolism , Syndrome , Serum Albumin/metabolism , Transaminases/metabolism , Triglycerides/metabolismالملخص
OBJETIVO: Descrever o perfil genético e metabólico de portadores da síndrome de Berardinelli-Seip (BSCL) acompanhados no Instituto da Criança do HC-FMUSP. SUJEITOS E MÉTODOS: Pacientes com as características clínicas da BSCL (n = 5), todas do sexo feminino, foram avaliadas com dosagens de glicose e insulina, lípides, leptina, enzimas hepáticas, análise de DNA, ultrassonografia abdominal. RESULTADOS: A deficiência de leptina e a hipertrigliceridemia foram constatadas nas cinco pacientes. Três evoluíram para diabetes melito (DM). Quatro tiveram mutação no gene AGPAT2 e uma no gene CAV1. CONCLUSÃO: As alterações metabólicas mais precoces foram a hipertrigliceridemia e a resistência insulínica, culminando no surgimento do DM à época da puberdade, sendo as mutações no gene AGPAT2 as mais frequentes em nossa casuística.
OBJECTIVE: To report the genetic and metabolic profile of patients with Berardinelli-Seip syndrome (BSCL) followed at Instituto da Criança, HC-FMUSP. SUBJECTS AND METHODS: Patients with clinical features of BSCL (n = 5), all female, were evaluated through serum levels of glucose, insulin, lipids, leptin, and liver enzymes. Abdominal sonography and DNA analysis were also performed. RESULTS: Leptin deficiency and hypertriglyceridemia were found in all the patients. Three progressed to diabetes mellitus. Four patients have mutations in AGPAT2 gene and one have a mutation in CAV1 gene. CONCLUSION: The earliest metabolic abnormalities were hypertriglyceridemia and insulin resistance, culminating in the onset of diabetes at the time of puberty. Mutations in the AGPAT2 gene were the most frequent in our patients.
الموضوعات
Adolescent , Child , Female , Humans , Young Adult , Lipodystrophy, Congenital Generalized/genetics , Lipodystrophy, Congenital Generalized/metabolism , /genetics , Caveolin 1/genetics , Diabetes Mellitus/etiology , Hypertriglyceridemia/diagnosis , Hypertriglyceridemia/pathology , Leptin/blood , Leptin/deficiency , Lipodystrophy, Congenital Generalized/complications , Mutation/genetics , Puberty/physiologyالملخص
To describe our 10-year experience in treating leptin-deficient humans. Three adults and one boy presented with childhood-onset morbid obesity, hypogonadism and family history of obesity and early death. Serum leptin was inappropriately low. A recessive C105T leptin gene mutation was identified. Metabolic and endocrine assessments were conducted, before and while on and off leptin. The adults' body mass index decreased from 51.2 ± 2.5 to 29.5 ± 2.8 kg/m². Serum lipids normalized, insulin resistance decreased, and one of the initially diabetic females became normoglycemic. Hypogonadotropic hypogonadism was reversed, and other changes were observed in the adrenal, sympathetic, somatotropic and thyroid functions. Leptin replacement therapy reverses endocrine and metabolic alterations associated with leptin deficiency. Some of these results may be extrapolated to other diseases.
Descrever nossa experiência de 10 anos tratando pacientes deficientes em leptina. Três adultos e um menino apresentaram obesidade mórbida com início na infância, hipogonadismo e história familiar de obesidade e morte precoce. A leptina sérica era inapropriadamente baixa. A mutação recessiva C105T no gene da leptina foi identificada. Avaliações metabólicas e endócrinas foram realizadas antes e durante o tratamento. O índice de massa corporal dos adultos baixou de 51,2 ± 2,5 para 29,5 ± 2,8 kg/m². Houve normalização dos lipídios séricos, a resistência insulínica diminuiu e a paciente que era diabética se tornou normoglicêmica. O hipogonadismo hipogonadotrópico foi revertido e outras alterações foram observadas nas funções adrenal, simpática, somatotrópica e tireoidiana. A reposição de leptina reverte as alterações endócrinas e metabólicas associadas com a deficiência de leptina. Alguns desses resultados podem ser extrapolados para outras doenças.