Suppression of high fat diet-induced liver cell injury by swim exercise / Supresión de lesión en células de hígado inducida por dieta alta en grasas con ejercicio de natación

SUMMARY: The rapid rise in obesity, particularly among children is a major public health concern that adversely affects vital organs including the liver. We sought to investigate the effect of exercise on the healing of liver cells from damage induced by high fat diet (HFD) in a rat model of hepatic steatosis. Rats were randomly divided into four groups (n=6 in each group); control group fed on a low fat diet (LFD), LFD plus exercise group (LFD+EX), model group fed on HFD, and swim exercise treated group (HFD+EX). Training swim exercise started from the 11th week up until the end of week 15. Liver index and body mass index (BMI) were determined, and harvested liver tissues were examined using basic histological staining and visualised under light microscopy. In addition, collected blood samples were assayed for biomarkers of liver injury. Histological images from the model group showed accumulation of lipid droplets in the hepatocytes (steatosis) and damaged liver cells that were inhibited by swimming exercise. Compared to control groups, HFD caused an increase in BMI and liver weight but not in liver index. In addition, HFD significantly (p<0.05) increased liver injury biomarkers; high-sensitivity C-reactive protein (hsCRP) and alkaline phosphatase (ALP) that were effectively (p<0.05) decreased by swimming exercise. Furthermore, a negative correlation between these biomarkers and the antioxidant and anti-inflammatory protein adiponectin was observed. Thus, HFD-induced hepatic steatosis is treated by swim exercise.

RESUMEN: El aumento de la obesidad, especialmente entre los niños, es un problema importante en la salud pública que afecta negativamente los órganos vitales, incluyendo el hígado. En este estudio se investigó el efecto del ejercicio en la curación de las células del hígado y el daño inducido por la dieta alta en grasas (HFD) en un modelo de rata de esteatosis hepática. Las ratas se dividieron aleatoriamente en cuatro grupos (n = 6 en cada grupo); grupo control, alimentado con una dieta baja en grasas (LFD); grupo de ejercicio LFD más (LFD + EX); grupo modelo alimentado con HFD; y grupo tratado con ejercicio de natación (HFD + EX). El entrenamiento con ejercicio de natación comenzó a partir de la semana 11 hasta el final de la semana 15. Se determinaron el índice hepático y el índice de masa corporal (IMC). Los tejidos hepáticos recolectados se examinaron mediante tinción histológica básica y se visualizaron con microscopía óptica. Además, se analizaron las muestras de sangre recogidas para identificar biomarcadores de lesión hepática. Las imágenes histológicas del grupo modelo mostraron acumulación de gotitas de lípidos en los hepatocitos (esteatosis) y células hepáticas dañadas que fueron inhibidas por el ejercicio de natación. En comparación con los grupos control, HFD causó un aumento en el IMC y el peso del hígado, pero no en el índice de hígado. Además, HFD aumentó significativamente (p <0.05) los biomarcadores de lesiones hepáticas; la proteína C reactiva de alta sensibilidad (hsCRP) y la fosfatasa alcalina (ALP) disminuyeron efectivamente (p <0.05) con el ejercicio de natación. Además, se observó una correlación negativa entre estos biomarcadores y la proteína antioxidante y antiinflamatoria adiponectina. Por lo tanto, la esteatosis hepática inducida por HFD puede ser tratada mediante el ejercicio de natación.

Potential role of marine algae extract on 3T3-L1 cell proliferation and differentiation: an in vitro approach

BACKGROUND: From ancient times, marine algae have emerged as alternative medicine and foods, contains the rich source of natural products like proteins, vitamins, and secondary metabolites, especially Chlorella vulgaris (C. vulgaris) contains numerous anti-inflammatory, antioxidants and wound healing substances. Type 2 diabetes mellitus is closely associated with adipogenesis and their factors. Hence, we aimed to investigate the chemical constituents and adipo-genic modulatory properties of C. vulgaris in 3T3-L1 pre-adipocytes. RESULTS: We analysed chemical constituents in ethanolic extract of C. vulgaris (EECV) by LC-MS. Results revealed that the EECV contains few triterpenoids and saponin compounds. Further, the effect of EECV on lipid accumulation along with genes and proteins expressions which are associated with adipogenesis and lipogenesis were evaluated using oil red O staining, qPCR and western blot techniques. The data indicated that that EECV treatment increased differentiation and lipid accumulation in 3T3-L1 cells, which indicates positive regulation of adipogenic and lipogenic activity. These increases were associated with up-regulation of PPAR-γ2, C/EBP-α, adiponectin, FAS, and leptin mRNA and protein expressions. Also, EECV treatments increased the concentration of glycerol releases as compared with control cells. Troglitazone is a PPAR-γ agonist that stimulates the PPAR-y2, adiponectin, and GLUT-4 expressions. Similarly, EECV treatments significantly upregulated PPAR-γ, adiponectin, GLUT-4 expressions and glucose utilization. Further, EECV treatment decreased AMPK-α expression as compared with control and metformin treated cells. CONCLUSION: The present research findings confirmed that the EECV effectively modulates the lipid accumulation and differentiation in 3T3-L1 cells through AMPK-α mediated signalling pathway.

Medidas comparativas do padrão morfométrico e perfil energético de éguas crioulas no terço final da gestação, com diferentes escores corporais / Comparative measurements of the morphometric pattern and energy profile of crioula mares in the final third of gestation, with different body condition scores

O objetivo deste estudo é comparar e descrever medidas morfométricas e perfil energético de éguas da raça Crioula no terço final da gestação. Foram utilizadas vinte éguas gestantes, divididas em 2 grupos conforme o escore corporal (BCS); o GrN (n=10), grupo de éguas com BCS 5 ou moderado, e GrO (n=10), grupo de éguas obesas ou BCS 9, analisadas conforme o mês de gestação (8º, 9º, 10º e 11º). Foram realizadas coletas de sangue e avaliações morfométricas mensais do peso corporal (PC), perímetro torácico (PT), perímetro abdominal (PA), altura da crista do pescoço (AC), espessura de gordura subcutânea na base da cauda (EGBC) e espessura de gordura retroperitoneal (EGRP). Para avaliar o perfil energético, foram mensurados os níveis séricos de triglicerídeos (TAG), ácidos graxos não esterificados (NEFA), Leptina (LEP) e Adiponectina. Os resultados das mensurações analisadas demonstraram no GrN incremento (P<0,05) nas variáveis PC, PT, PA e NEFA do 8º para o 9º mês de gestação. Porém, na variável EGBC, o aumento (P<0,01) foi observado entre o 8º e o 11º mês avaliado. No GrO, o PA demonstrou incremento (P<0,01) entre o 8º e o 11º mês de gestação, e a LEP apresentou aumento (P<0,05) entre o 10º e o 11º mês. Quando os dois grupos foram comparados, as variáveis PC, PT, EGBC e EGRP apresentaram diferença (P<0,05) em todos os meses avaliados. Na análise de correlação, o BCS associou-se com o PC (r=0,691; P<0,0001), PT (r=0,705; P<0,0001), EGBC (r=0,701; P<0,0001), EGRP (r=0,627; P<0,0001), níveis séricos de LEP (r=0,426; P<0,0001) e Adiponectina (r=0,217; P<0,05). Dessa forma, pode-se afirmar que a gordura corporal aumentou progressivamente nas éguas do GrN, diferentemente do observado no GrO. Através dos métodos utilizados neste trabalho, torna-se possível a avaliação prática da morfometria das éguas da raça Crioula no terço final da gestação, possibilitando de forma objetiva a identificação de indivíduos obesos.

The aim of this study was to compare and describe morphometric measurements and energy profile of the Crioula breed mares at the end of gestation. Twenty pregnant mares were used and separated into 2 groups according to body condition score (BCS). The GrN (n= 10) group of mares with BCS 5 or moderate and GrO (n = 10) group of obese mares or BCS 9 were analyzed according to the month of pregnancy (8th, 9th, 10th and 11th). Blood samples and monthly morphometric evaluation were performed for body weight (PC), heart girth (PT), waist circumference (PA), height of the crest of the neck (AC), thickness subcutaneous fat on the base of the tail (EGBC) and retroperitoneal (EGRP). To evaluate the energy profile serum levels of triglycerides (TAG), non-esterified fatty acids (NEFA), leptin (LEP) and adiponectin were measured. The results of the analyzed measurements showed an increase in GrN (P<0.05) in the PC, PT, PA and NEFA variables between the 8th and 9th month of pregnancy. Although there was an increase (P<0.01) between the 8th and 11th month of pregnancy in the EGBC variable, in GrO the PA was increased (P<0.01) between the 8th and 11th month of pregnancy and the LEP showed an increase (P<0.05) between the 10th and 11th month. When both groups were compared, the variables PC, PT, EGBC and EGRP showed differences (P<0.05) in all months evaluated. In the correlation the BCS was associated with PC (r=0.691, P<0.0001), PT (r=0.705, P<0.0001), EGBC (r=0.701, P<0.0001), EGRP (r=0.627, P<0.0001), serum leptin levels (r=0.426, P<0.0001) and adiponectin (r=0.217, P<0.05). Thus, it can be stated that the body fat increased progressively in mares GrN, different from that observed in GrO. Therefore, by the methods used in this study, the morphometry practical evaluation of the Crioula breed mares in the last third of gestation became possible, allowing an objective way to identify obese individuals.

Triiodothyronine modulates the expression of leptin and adiponectin in 3T3-L1 adipocytes / Triiodotironina modula a expressão de leptina e adiponectina em adipócitos 3T3-L1

Objective To study the effect of different doses of triiodothyronine on gene expression of the adipokines leptin and adiponectin, at different times, and to evaluate the difference in expression between the two adipokines in each group. Methods 3T3-L1 adipocytes were incubated with triiodothyronine at physiological dose (10nM) and supraphysiological doses (100nM or 1,000nM), or without triiodothyronine (control, C) for 0.5, 6, or 24 hours. Leptin and adiponectin mRNA was detected using real-time polymerase chain reaction (RT-PCR). One-way analyses of variance, Tukey’s test or Student’s t test, were used to analyze data, and significance level was set at 5%. Results Leptin levels decreased in the 1,000nM-dose group after 0.5 hour. Adiponectin levels dropped in the 10nM-dose group, but increased at the 100nM dose. After 6 hours, both genes were suppressed in all hormone concentrations. After 24 hours, leptin levels increased at 10, 100 and 1,000nM groups as compared to the control group; and adiponectin levels increased only in the 100nM group as compared to the control group. Conclusion These results demonstrated fast actions of triiodothyronine on the leptin and adiponectin expression, starting at 0.5 hour, at a dose of 1,000nM for leptin and 100nM for adiponectin. Triiodothyronine stimulated or inhibited the expression of adipokines in adipocytes at different times and doses which may be useful to assist in the treatment of obesity, assuming that leptin is increased and adiponectin is decreased, in obesity cases. .

Objetivo Examinar o efeito de diferentes doses de triiodotironina sobre a expressão gênica das adipocinas leptina e adiponectina, em diferentes períodos de tempo, além de avaliar a diferença de expressão entre as duas adipocinas em cada grupo. Métodos Adipócitos 3T3-L1 foram incubados com triiodotironina nas doses fisiológica (10nM) e suprafisiológicas (100nM ou 1.000nM), ou na ausência de triiodotironina (controle, C) durante 0,5, 6 ou 24 horas. O mRNA das adipocinas foi analisado em tempo real, utilizando a reação em cadeia de polimerase. Para as análises dos dados, foi utilizada a análise de variância, complementada com o teste de Tukey, ou o teste t de Student com 5% de significância. Resultados Os níveis de leptina diminuíram no grupo com dose de 1.000nM em 0,5 hora. A adiponectina também diminuiu no grupo com dose de 10nM, porém se elevou com a dose de 100nM. Após 6 horas, ambos os genes foram suprimidos em todas concentrações de hormônio. Em 24 horas, os níveis de leptina foram elevados em 10, 100 e 1.000nM, em relação ao grupo controle. No que concerne à adiponectina, observou-se aumento apenas no grupo cuja dose foi de 100nM, em comparação ao controle. Conclusão Foram demonstradas ações rápidas da triiodotironina sobre a expressão da leptina e da adiponectina, iniciando em 0,5 hora na dose de 1.000nM, para a primeira, e na dose de 100nM, para a segunda. A triiodotironina estimulou ou inibiu a expressão de adipocinas em adipócitos em diferentes tempos e doses, o que pode auxiliar no tratamento da obesidade, levando em consideração que, nesta, a leptina está aumentada e adiponectina, diminuída. .

Imunoexpressão hepática de marcadores inflamatórios e de resistência insulínica na esteato-hepatite não alcóolica / Immunohistochemistry pattern of hepatic inflammatory and insulin resistance markers in experimental model of nonalcoholic steatohepatitis

Introduction: The pathophysiology of nonalcoholic steatohepatitis (NAS) includes, basically, insulin resistance, inflammation and oxidative stress. Thus, a study of immunostaining for liver insulin, adiponectin, tumor necrosis factor alpha (TNF-α), and inducible nitric oxide synthase (iNOS) receptors was conducted. Objective: To expand the knowledge about the pathophysiological and molecular mechanisms underlying the experimental model of steatohepatitis in rats fed a high-fat diet. Method: Twenty Wistar rats were divided into two groups: G1 (control, fed a standard diet), and G2 (fed a high-fat diet containing 58% of energy derived from fat, 18% from protein and 24% from carbohydrate). After eight weeks the animals were sacrificed. Blood glucose, insulin, total cholesterol, high-density lipoprotein (HDL), the very low-density lipoproteins (VLDL), triglycerides, aspartate aminotransferase (AST), alanine aminotransferase (ALT), alkaline phosphatase (ALP) and gamma-glutamyl transferase (GGT) were determined. The liver tissue was submitted to histopathological analysis, using a NAS score. In immunohistochemistry, we studied the expression of the insulin receptor, adiponectin, TNF-α and iNOS by tissue microarray method. Results and conclusion: There was marked cytoplasmic immunostaining for TNF-α and iNOS mediators in the group on a fat diet. Regarding insulin and adiponectin molecular markers, a reduction of cytoplasmic immunoreactivity of these antigens was observed in the group on a fat diet, reflecting, respectively, the state of hepatocellular inflammation (steatohepatitis) and insulin resistance in this experimental model of fat liver disease...

Introdução: Os mecanismos fisiopatológicos da esteato-hepatite não alcoólica incluem basicamente resistência insulínica, processo inflamatório e estresse oxidativo. Desta forma, um estudo sobre o padrão de imunoexpressão hepática para receptores de insulina, adiponectina, fator de necrose tumoral alfa (TNF-α) e sintase indutível do óxido nítrico (iNOS) foi conduzido. Objetivo: Ampliar os conhecimentos sobre os mecanismos moleculares subjacentes, em modelo experimental de esteato-hepatite. Método: Vinte ratos Wistar com dois meses de idade, pesando de 250 a 300 mg foram subdivididos em dois grupos: G1 (controle normal, submetido à dieta padrão) e G2 (grupo-controle, submetido à dieta hiperlipídica contendo 58% de energia derivada de gorduras, 18% de proteínas e 24% de carboidratos). Após oito semanas, os animais foram sacrificados; o sangue, submetido à análise bioquímica; e o fígado, removido e fixado em formalina tamponada e emblocado em parafina para estudo histopatológico. Para estudo imuno-histoquímico, foi utilizada a técnica de microarranjo de tecido. As lâminas obtidas foram submetidas à incubação com os anticorpos contra adiponectina, receptor de insulina, TNF-α e iNOS. Resultados e conclusão: Observou-se marcada imunoexpressão citoplasmática para os mediadores TNF-α e iNOS no grupo submetido à dieta hiperlipídica. No que diz respeito aos marcadores moleculares insulina e adiponectina, observou-se uma redução da imunoexpressão citoplasmática desses anticorpos no grupo submetido à dieta hiperlipídica, traduzindo, respectivamente, o estado de inflamação hepatocelular (esteato-hepatite) e resistência insulínica, desenvolvidos nesse modelo experimental de doença hepática gordurosa...

Pathophysiological Role of Hormones and Cytokines in Cancer Cachexia

We investigated the role of fasting hormones and pro-inflammatory cytokines in cancer patients. Hormones (ghrelin, adiponectin, and leptin) and cytokines (TNF-alpha, IFN-gamma, and IL-6) were measured by ELISA or RIA in lung cancer and colorectal cancer patients before the administration of cancer therapy, and measurements were repeated every 2 months for 6 months. From June 2006 to August 2008, 42 patients (19 with colorectal cancer and 23 with lung cancer) were enrolled. In total, 21 patients were included in the cachexia group and the others served as a comparison group. No significant difference in the initial adiponectin, ghrelin, TNF-alpha, IFN-gamma, or IL-6 level was observed between groups, although leptin was significantly lower in cachectic patients than in the comparison group (15.3 +/- 19.5 vs 80.9 +/- 99.0 pg/mL, P = 0.007). During the follow-up, the patients who showed a > 5% weight gain had higher ghrelin levels after 6 months. Patients exhibiting elevated IL-6 levels typically showed a weight loss > 5% after 6 months. A blunted adiponectin or ghrelin response to weight loss may contribute to cancer cachexia and IL-6 may be responsible for inducing and maintaining cancer cachexia.

Relação adiponectina/leptina como método para avaliação da sensibilidade insulínica / Adiponectin/leptin ratio as an evaluation method to insulin sensitivity

Nos últimos anos, ocorreram mudanças na compreensão da fisiopatologia de doenças que cursam com inflamação subclínica, entre elas a obesidade e o diabetes mellitus tipo 2, condições que têm como substrato comum a resistência insulínica. A avaliação da sensibilidade à insulina (SI) é realizada por métodos indiretos como o cálculo do índice HOMA-IR que apresenta boa correlação com o clamp euglicêmico hiperinsulinêmico, considerado padrão ouro. Recentemente, a relação entre adiponectina/leptina (A/L), citocinas envolvidas na fisiopatologia da síndrome metabólica, tem sido proposta como método para avaliação da SI. Revisamos estudos sobre a relação A/L para avaliação da SI e os dados mostraram que, comparada ao HOMA-IR, a relação A/L constitui um marcador confiável de SI em pacientes com diabetes mellitus tipo 2 e em não diabéticos. Estes achados permitem sugerir a relação A/L como método de avaliação da SI, mas necessitam de confirmação em estudos com maior número de indivíduos.

The knowledge of the pathophysiology of inflammatory diseases coursing with insulin resistance such as obesity and type 2 diabetes has changed. The assessment of insulin sensitivity (IS) is carried out through indirect methods such as the HOMA-IR which shows a good correlation with the euglycemic hyperinsulinemic clamp, the gold standard for IS. Recently, the ratio between adiponectin/leptin (A/L), cytokines involved in the pathophysiology of the metabolic syndrome, has been proposed as an alternative method for the assessment of the IS. In this paper the authors reviewed studies that examined the A/L ratio as a tool to estimate IS and the data showed that compared to HOMA-IR, the A/L ratio is a reliable marker for IS in obese patients with and without type 2 diabetes. These findings suggest that the ratio A/L could be used as an evaluation method to estimate IS but should be confirmed in studies with high number of individuals.

Adiponectin muitimers in patients with Barrett's Oesophagus

Barrett's oesophagus is associated with abdominal obesity. Adiponectin is a peptide that is secreted from adipocytes and circulates in three multimeric forms: low molecular weight [LMW], middle molecular weight[MMW], and high molecular weight [HMW]. The anti-inflammatory effects of adiponectin are specific to individual muitimers, with the LMW being most anti-inflammatory. We investigated the possibility that circulating levels of adiponectin and its multimers would be associated with the risk of Barrett's oesophagus. This study comprised patients diagnosed to have Barrett's oesophagus, and control subjects diagnosed to have gastrooesophageal reflux disease [GORD], all were diagnosed in the gastroenterology and endoscopy unit, King Fahd hospital, Riyadh, Saudi Arabia. Plasma adiponectin levels and its multimers were evaluated for patients with Barrett's oesophagus and controls with GORD. There were 120 cases of Barrett's oesophagus and 250 GORD controls. Total adiponectin was not significantly associated with Barrett's oesophagus; low levels of LMW adiponectin and low LMW/total ratio were significantly present in patients with Barrett's oesophagus. Low levels of LMW adiponectin are significantly associated with Barrett's oesophagus

Relationship of body composition and circulatory adiponectin to bone mineral density in young premenopausal women.

OBJECTIVE: Adiponectin is a recently discovered hormone secreted by adipocytes. Adiponectin plays an important role in the regulation of insulin sensitivity as well as the propensity to inflammation and atherosclerosis. In the present study, the authors explore the relationship between adiponectin and bone mass in premenopausal women. The relationship of fat mass compared to lean body mass to bone mass was also investigated MATERIAL AND METHOD: Two hundred premenopausal women aged between 20 and 40 years were studied. Bone mineral density (BMD) was measured at L2-4 and femoral neck by dual-energy X-ray absorptiometry (DEXA). Serum adiponectin concentrations were measured by radioimmunoassay. RESULTS: At the lumbar spines, factors associated with BMD were age (p < 0.01) and lean body mass (p < 0.001). No independent association with fat mass was demonstrated Likewise, at the femoral neck, only lean body mass was related to BMD (p < 0. 01). In terms of the relation of serum adiponectin to BMD, no association of serum adiponectin to BMD at the lumbar spines or femoral neck was found CONCLUSION: Altogether, the present findings do not suggest the independent role of adiponectin in the accrual of bone mass in females, although such a role still cannot be excluded in men or postmenopausal women.