RESUMEN
Peripheral action of irisin improves glucose homeostasis and increases energy expenditure, with no data on a central role of irisin in metabolism. These studies sought to examine 1) presence of irisin in human cerebrospinal fluid (CSF) and banked human hypothalamic tissue, 2) serum irisin in maternal subjects across varying adiposities with or without gestational diabetes (GDM), and 3) their respective neonate offspring. CSF, serum, and neonatal cord serum were collected from 91 pregnant women with and without GDM attending for an elective cesarean section [body mass index (BMI): 37.7 ± 7.6 kg/m(2); age: 32 ± 8.3 yr]. Irisin was assessed by ELISA and correlated with biochemical and anthropometric data. Irisin expression was examined in human hypothalamus by immunohistochemical staining. Serum irisin in pregnant women was significantly lower in nonobese compared with obese and GDM subjects, after adjusting for BMI, lipids, and glucose. Irisin was present in neonatal cord serum (237 ± 8 ng/ml) and maternal CSF (32 ± 1.5 ng/ml). CSF irisin correlated positively with serum irisin levels from nonobese and obese pregnant women (P < 0.01), with CSF irisin significantly raised in GDM subjects (P < 0.05). Irisin was present in human hypothalamic sections in the paraventricular neurons, colocalized with neuropeptide Y. Irisin was detectable in CSF and in paraventricular neurons. Maternal serum irisin was lower in nonobese pregnant women after adjusting for BMI and a number of metabolic parameters. These studies indicate that irisin may have a central role in metabolism in addition to the known peripheral role. Further studies investigating the central action of irisin in human metabolic disease are required.
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Adiposidad/fisiología , Diabetes Gestacional/metabolismo , Fibronectinas/metabolismo , Hipotálamo/metabolismo , Obesidad/metabolismo , Adulto , Biomarcadores/líquido cefalorraquídeo , Biomarcadores/metabolismo , Diabetes Gestacional/líquido cefalorraquídeo , Femenino , Fibronectinas/líquido cefalorraquídeo , Humanos , Neuronas/metabolismo , Neuropéptido Y/metabolismo , Obesidad/líquido cefalorraquídeo , EmbarazoRESUMEN
BACKGROUND: Irisin, a novel myokine, increases energy expenditure and glucose tolerance and, thus, improves carbohydrate homeostasis in humans. This hormone has potential therapeutic applications for weight loss and improvement in insulin resistance in subjects with obesity and diabetes mellitus type 2 (T2DM). In this cross-sectional study, we aimed to associate circulating levels of irisin and several anthropometric and metabolic parameters among Arab children. METHODS: A cohort of 153 Saudi children, 81 boys [age: 12·4 ± 3·2 years; BMI: 19·5 ± 5·9 kg/m(2) ] and 72 girls: [age: 12·9 ± 3·2 years; BMI: 20·6 ± 5·2], were examined. Anthropometry was obtained, and fasted bloods were collected for biochemical analyses. Irisin was assessed by a specific enzyme-linked immunosorbent assay (ELISA). RESULTS: Girls had higher circulating irisin levels than boys (P = 0·04). There were several significant correlations between circulating irisin and fasting blood glucose (FBG) (r = -0·35, P < 0·001), sagittal abdominal diameter (SAD) (r = -0·34, P < 0·001) and HDL cholesterol (r = 0·17, P = 0·04) across the entire cohort studied. Notably in girls, but not in boys, HOMA-IR correlated negatively with irisin levels (r = -0·32, P = 0·02), as previously noted in adults. FBG was a significant predictor of circulating irisin (R(2) = 0·16) followed by SAD. In multivariate linear regression analysis, after controlling for potential confounders such as gender, age and BMI, irisin levels were independently associated with FBG (ß = -0·34, P = 0·01), particularly in girls. CONCLUSION: Serum irisin levels were higher in girls than in boys and correlated negatively with HOMA-IR. They were also independently associated with FBG predominantly in girls, suggesting that this hormone may play a crucial role in glucose metabolism from an early age.
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Glucemia/metabolismo , Fibronectinas/metabolismo , Biomarcadores/metabolismo , Niño , Estudios Transversales , Ensayo de Inmunoadsorción Enzimática , Femenino , Homeostasis/fisiología , Humanos , Masculino , Factores SexualesRESUMEN
PURPOSE OF REVIEW: This article reviews the evidence linking gut bacteria, endotoxin, and its circulating levels with inflammatory induced obesity and metabolic disease (metabolic endotoxaemia). RECENT FINDINGS: Gut flora analyses have allowed gut microbiota signatures (GMS) to be observed in animal studies of obesity/metabolic disease. In these studies, specific GMS result in a change in obesity and metabolic disease state whereas in humans, analysis remains unclear. Serum studies, examining metabolic endotoxaemia as a biomarker, appear to link long-term cardiovascular disease and type 2 diabetes mellitus (T2DM) through activation of inflammatory pathways. More recent studies note the importance of diet, which shows the dramatic rise in endotoxin following acute or long-term high-fat diet, with the effects exacerbated in T2DM. SUMMARY: Gut flora appears to act as an important determinant in the pathogenesis of inflammatory induced obesity/T2DM. Endotoxin may act as the systemic insult, impacted by a high-fat diet, which may regulate this effect, combined with an altered GMS. As such, clinical and dietary intervention to affect this process - on the gut flora, the 'leaky' mucosal membrane and endotoxin coupled lipid absorption or removal of circulating endotoxin - could reduce the progression of inflammatory induced metabolic disease.
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Dieta Alta en Grasa/efectos adversos , Endotoxemia/metabolismo , Tracto Gastrointestinal/microbiología , Inflamación/microbiología , Animales , Bacterias/crecimiento & desarrollo , Bacterias/inmunología , Bacterias/patogenicidad , Biomarcadores/metabolismo , Diabetes Mellitus Tipo 2/patología , Endotoxemia/inmunología , Endotoxemia/microbiología , Tracto Gastrointestinal/inmunología , Tracto Gastrointestinal/metabolismo , Humanos , Inflamación/patología , Mucosa Intestinal/inmunología , Mucosa Intestinal/microbiología , Metabolismo de los Lípidos , Lipopolisacáridos/inmunología , Metagenoma , Obesidad/inmunología , Obesidad/microbiología , Obesidad/patología , Factores de RiesgoRESUMEN
OA (osteoarthritis) is a degenerative condition associated with obesity. A number of metabolic explanations have been proposed to explain the association between obesity and OA in non-weight-bearing joints; however, none of these hypotheses have been demonstrated empirically. In the present Hypothesis article, we recognize that obesity is associated with compromised gut mucosa, translocation of microbiota and raised serum LPS (lipopolysaccharide). The consequent activation of the innate immune response leads to increased serum titres of inflammatory mediators in obese patients, with both local and systemic markers of inflammation associated with onset and progression of OA. Furthermore, a number of workers have shown that articular cartilage repair is impaired by a range of inflammatory mediators, both in vitro and in vivo. We propose that metabolic endotoxaemia, caused by impaired gastric mucosa and low-grade chronic inflammation, may contribute to the onset and progression of OA in obese patients. This may account for the association between obesity and OA at non-weight-bearing joints which cannot be explained by biomechanical factors.
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Endotoxemia/complicaciones , Obesidad/complicaciones , Osteoartritis/etiología , Traslocación Bacteriana , Fenómenos Biomecánicos , Cartílago Articular/metabolismo , Cartílago Articular/patología , Cartílago Articular/fisiopatología , Endotoxemia/sangre , Endotoxemia/fisiopatología , Humanos , Inflamación/sangre , Inflamación/complicaciones , Inflamación/fisiopatología , Mediadores de Inflamación/sangre , Mediadores de Inflamación/metabolismo , Mucosa Intestinal/metabolismo , Mucosa Intestinal/microbiología , Mucosa Intestinal/patología , Lipopolisacáridos/sangre , Modelos Biológicos , Obesidad/sangre , Obesidad/fisiopatología , Osteoartritis/sangre , Osteoartritis/fisiopatología , Factores de Riesgo , Soporte de PesoRESUMEN
INTRODUCTION: Chronic low-grade inflammation is a significant factor in the development of obesity associated diabetes. This is supported by recent studies suggesting endotoxin, derived from gut flora, may be key to the development of inflammation by stimulating the secretion of an adverse cytokine profile from adipose tissue. AIMS: The study investigated the relationship between endotoxin and various metabolic parameters of diabetic patients to determine if anti-diabetic therapies exerted a significant effect on endotoxin levels and adipocytokine profiles. METHODS: Fasting blood samples were collected from consenting Saudi Arabian patients (BMI: 30.2 +/- (SD)5.6 kg/m2, n = 413), consisting of non-diabetics (ND: n = 67) and T2DM subjects (n = 346). The diabetics were divided into 5 subgroups based on their 1 year treatment regimes: diet-controlled (n = 36), metformin (n = 141), rosiglitazone (RSG: n = 22), a combined fixed dose of metformin/rosiglitazone (met/RSG n = 100) and insulin (n = 47). Lipid profiles, fasting plasma glucose, insulin, adiponectin, resistin, TNF-alpha, leptin, C-reactive protein (CRP) and endotoxin concentrations were determined. RESULTS: Regression analyses revealed significant correlations between endotoxin levels and triglycerides (R2 = 0.42; p < 0.0001); total cholesterol (R2 = 0.10; p < 0.001), glucose (R2 = 0.076; p < 0.001) and insulin (R2 = 0.032; p < 0.001) in T2DM subjects. Endotoxin showed a strong inverse correlation with HDL-cholesterol (R2 = 0.055; p < 0.001). Further, endotoxin levels were elevated in all of the treated diabetic subgroups compared with ND, with the RSG treated diabetics showing significantly lower endotoxin levels than all of the other treatment groups (ND: 4.2 +/- 1.7 EU/ml, RSG: 5.6 +/- 2.2 EU/ml). Both the met/RSG and RSG treated groups had significantly higher adiponectin levels than all the other groups, with the RSG group expressing the highest levels overall. CONCLUSION: We conclude that sub-clinical inflammation in T2DM may, in part, be mediated by circulating endotoxin. Furthermore, that whilst the endotoxin and adipocytokine profiles of diabetic patients treated with different therapies were comparable, the RSG group demonstrated significant differences in both adiponectin and endotoxin levels. We confirm an association between endotoxin and serum insulin and triglycerides and an inverse relationship with HDL. Lower endotoxin and higher adiponectin in the groups treated with RSG may be related and indicate another mechanism for the effect of RSG on insulin sensitivity.
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Diabetes Mellitus Tipo 2/sangre , Diabetes Mellitus Tipo 2/tratamiento farmacológico , Endotoxinas/sangre , Hipoglucemiantes/uso terapéutico , Adulto , Anciano , Anciano de 80 o más Años , Glucemia/efectos de los fármacos , Glucemia/metabolismo , Femenino , Humanos , Hipoglucemiantes/farmacología , Masculino , Persona de Mediana Edad , Adulto JovenRESUMEN
CONTEXT: The adipokine leptin has critical importance in central appetite regulation. In contrast to some suggestion of adiponectin influencing energy homeostasis in rodents, there is no evidence for adiponectin or resistin entering the human blood-brain barrier. OBJECTIVE: The objective was to establish the presence of adiponectin or resistin in human cerebrospinal fluid (CSF) and to compare their distribution with leptin. Furthermore, we wished to examine the expression of the adiponectin receptors 1 and 2 (AdipR1, AdipR2) in the human hypothalamus. METHODS: For this purpose, serum and CSF samples were collected from 20 men and 19 women matched for age [men, 69.8 +/- 8.6 yr (mean +/- SD); women, 69.4 +/- 4.3 yr] and BMI (men, 29.4 +/- 3.4 kg/m(2); women, 27.3 +/- 4.8 kg/m(2)) undergoing elective surgery under spinal anesthesia. RESULTS: Adiponectin was identified in CSF with levels 1000-fold less than serum, whereas resistin and leptin levels were 100-fold less. Unlike their serum levels, adiponectin CSF levels showed no gender difference or correlation with insulin resistance, which is similar to resistin CSF levels. The adiponectin and leptin CSF/serum ratios in our study exhibit the same pattern of gender-specific BMI association with inverse correlation in women (r = -0.61; P = 0.02) and no correlation in men (r = 0.026; P = not significant). Furthermore, immunostaining established AdipR1 and -2 in the hypothalamus and increased AdipR2 expression in the paraventricular nucleus, which is involved in energy regulation. CONCLUSION: In summary, our findings show both the presence of adiponectin and resistin in human CSF, with no effect of insulin resistance on CSF levels. The CSF entry of adiponectin and leptin in women appears to be impaired in obesity.
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Adiponectina/líquido cefalorraquídeo , Hipotálamo/metabolismo , Receptores de Superficie Celular/metabolismo , Resistina/líquido cefalorraquídeo , Adiponectina/sangre , Anciano , Índice de Masa Corporal , Estudios de Casos y Controles , Femenino , Humanos , Inmunohistoquímica , Leptina/sangre , Leptina/líquido cefalorraquídeo , Masculino , Persona de Mediana Edad , Receptores de Adiponectina , Resistina/sangreRESUMEN
CONTEXT: Obesity-associated inflammation is a contributory factor in the pathogenesis of type 2 diabetes mellitus (T2DM); the mechanisms underlying the progression to T2DM are unclear. The adipokine resistin has demonstrated proinflammatory properties in relation to obesity and T2DM. OBJECTIVES: The objectives of this study were to characterize resistin expression in human obesity and address the role of resistin in the innate immune pathway; to examine the influence of lipopolysaccharide, recombinant human resistin (rhResistin), insulin, and rosiglitazone in human adipocytes; and, finally, to analyze the effect of rhResistin on the expression of components of the nuclear factor-kappaB pathway and insulin signaling cascade. METHODS: Abdominal sc adipose tissue was obtained from patients undergoing elective liposuction surgery (n = 35; age, 36-49 yr; body mass index, 26.5 +/- 5.9 kg/m2). Isolated adipocytes were cultured with rhResistin (10-50 ng/ml). The level of cytokine secretion from isolated adipocytes was examined by ELISA. The effect of rhResistin on protein expression of components of the innate immune pathway was examined by Western blot. RESULTS: In vitro studies demonstrated that antigenic stimuli increase resistin secretion (P < 0.001) from isolated adipocytes. Proinflammatory cytokine levels were increased in response to rhResistin (P < 0.001); this was attenuated by rosiglitazone (P < 0.01). When examining components of the innate immune pathway, rhResistin stimulated Toll-like receptor-2 protein expression. Similarly, mediators of the insulin signaling pathway, phosphospecific c-Jun NH2-terminal kinase (JNK) 1 and JNK2, were up-regulated in response to rhResistin. CONCLUSION: Resistin may participate in more than one mechanism to influence proinflammatory cytokine release from human adipocytes, potentially via the integration of nuclear factor-kappaB and JNK signaling pathways.
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Adipocitos/inmunología , Inmunidad Innata/efectos de los fármacos , Resistina/farmacología , Transducción de Señal/efectos de los fármacos , Adipocitos/efectos de los fármacos , Adulto , Índice de Masa Corporal , Humanos , Interleucina-6/metabolismo , Persona de Mediana Edad , FN-kappa B/fisiología , Proteínas Recombinantes/farmacología , Piel/citología , Piel/inmunología , Receptor Toll-Like 2/análisis , Receptor Toll-Like 4/análisis , Factor de Necrosis Tumoral alfa/metabolismoRESUMEN
OBJECTIVE: Calcitonin gene-related peptide (CGRP) is a vasoactive, proinflammatory neuropeptide implicated in the pathogenesis of cardiovascular disease. Elevated CGRP levels during hot flushes and pregnancy suggest that reproductive hormones may influence CGRP secretion. CGRP and the related protein adrenomedullin (ADM) may function through adipose tissue-mediated effects, since adipose tissue is an important site of cytokine production and the main site for estrogen production after menopause. This study examined mRNA and protein expression of CGRP, ADM, and the receptor activity-modifying proteins and the effects of menopausal status in human adipose tissue. DESIGN: Protein/mRNA levels were determined in adipose tissue biopsy samples collected from premenopausal (n = 22: follicle-stimulating hormone, <20 IU/L; estradiol [mean +/- SEM], 434.5 +/- 87.81 pmol/L) and postmenopausal (n = 25: follicle-stimulating hormone, >20 IU/L; estradiol, 43.4 +/- 6.95 pmol/L) women. RESULTS: Our studies determined that CGRP, ADM, and receptor activity-modifying proteins were expressed in abdominal fat, adipocytes, and preadipocytes. CGRP and ADM mRNA levels were increased in abdominal subcutaneous fat in postmenopausal women compared with premenopausal women (betaCGRP: premenopause Delta cycle threshold [Ct], 31.07 +/- 0.28 vs postmenopause DeltaCt, 30.35 +/- 0.17, P = 0.035; ADM: premenopause subcutaneous fat DeltaCt, 12.41 +/- 0.2 vs postmenopause subcutaneous fat DeltaCt, 11.55 +/- 0.14, P < 0.001) with CGRP differentially expressed in subcutaneous and omental depots. CGRP protein expression was higher in postmenopausal women (P < 0.05) in both fat depots. CONCLUSIONS: Our findings suggest that adipose tissue represents an important site for CGRP and ADM production and that menopause status alters their expression in abdominal fat. This offers a potential mechanism to explain the role of CGRP in menopausal vasomotor symptoms and the increased risk of cardiovascular disease in postmenopausal women.
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Tejido Adiposo/metabolismo , Adrenomedulina/metabolismo , Péptido Relacionado con Gen de Calcitonina/metabolismo , Moduladores de los Receptores de Estrógeno/metabolismo , Menopausia , Adulto , Estudios de Casos y Controles , Cartilla de ADN , Femenino , Regulación de la Expresión Génica , Humanos , Persona de Mediana Edad , Proteínas/análisis , ARN Mensajero/análisis , Reacción en Cadena de la Polimerasa de Transcriptasa InversaRESUMEN
INTRODUCTION: Inflammation contributes to cardiovascular disease and is exacerbated with increased adiposity, particularly omental adiposity; however, the role of epicardial fat is poorly understood. METHODS: For these studies the expression of inflammatory markers was assessed in epicardial fat biopsies from coronary artery bypass grafting (CABG) patients using quantitative RT-PCR. Further, the effects of chronic medications, including statins, as well as peri-operative glucose, insulin and potassium infusion, on gene expression were also assessed. Circulating resistin, CRP, adiponectin and leptin levels were determined to assess inflammation. RESULTS: The expression of adiponectin, resistin and other adipocytokine mRNAs were comparable to that in omental fat. Epicardial CD45 expression was significantly higher than control depots (p < 0.01) indicating significant infiltration of macrophages. Statin treated patients showed significantly lower epicardial expression of IL-6 mRNA, in comparison with the control abdominal depots (p < 0.001). The serum profile of CABG patients showed significantly higher levels of both CRP (control: 1.28 +/- 1.57 microg/mL vs CABG: 9.11 +/- 15.7 microg/mL; p < 0.001) and resistin (control: 10.53 +/- 0.81 ng/mL vs CABG: 16.8 +/- 1.69 ng/mL; p < 0.01) and significantly lower levels of adiponectin (control: 29.1 +/- 14.8 microg/mL vs CABG: 11.9 +/- 6.0 microg/mL; p < 0.05) when compared to BMI matched controls. CONCLUSION: Epicardial and omental fat exhibit a broadly comparable pathogenic mRNA profile, this may arise in part from macrophage infiltration into the epicardial fat. This study highlights that chronic inflammation occurs locally as well as systemically potentially contributing further to the pathogenesis of coronary artery disease.
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Tejido Adiposo/química , Enfermedad de la Arteria Coronaria/genética , Enfermedad de la Arteria Coronaria/metabolismo , Citocinas/análisis , Pericardio/química , Adiponectina/análisis , Adiponectina/sangre , Adiponectina/genética , Tejido Adiposo/patología , Tejido Adiposo/fisiopatología , Proteína C-Reactiva/análisis , Puente de Arteria Coronaria , Enfermedad de la Arteria Coronaria/tratamiento farmacológico , Enfermedad de la Arteria Coronaria/patología , Citocinas/genética , Perfilación de la Expresión Génica , Regulación de la Expresión Génica/efectos de los fármacos , Glucosa/administración & dosificación , Humanos , Inhibidores de Hidroximetilglutaril-CoA Reductasas/uso terapéutico , Insulina/administración & dosificación , Interleucina-6/análisis , Interleucina-6/genética , Grasa Intraabdominal/fisiopatología , Leptina/sangre , Leptina/genética , Antígenos Comunes de Leucocito/análisis , Antígenos Comunes de Leucocito/genética , Macrófagos/química , Macrófagos/patología , Persona de Mediana Edad , Pericardio/patología , Pericardio/fisiopatología , Reacción en Cadena de la Polimerasa , Potasio/administración & dosificación , ARN Mensajero/análisis , ARN Mensajero/genética , Resistina/análisis , Resistina/sangre , Resistina/genéticaRESUMEN
Lipolysis is an important process determining fuel metabolism, and insulin regulates this process in adipose tissue. The aim of this study was to investigate the long-term effects of insulin, an insulin enhancer (rosiglitazone [RSG]), and insulin in combination with RSG on the regulation of lipolysis and lipogenesis in human abdominal subcutaneous fat. Lipolysis and lipogenesis were assessed by protein expression studies of hormone-sensitive lipase (HSL) (84 kDa) and lipoprotein lipase (LPL) (56 kDa), respectively. In addition, lipolytic rate was assessed by glycerol release assay and tumor necrosis factor (TNF)-alpha release measured by enzyme-linked immunosorbent assay (n = 12). In subcutaneous adipocytes, increasing insulin doses stimulated LPL expression, with maximal stimulation at 100 nmol/l insulin (control, 1.0 +/- 0.0 [mean +/- SE, protein expression relative to control]; 1 nmol/l insulin, 0.87 +/- 0.13; 100 nmol/l insulin, 1.68 +/- 0.19; P < 0.001). In contrast, insulin at the 100 nmol/l dose reduced the expression of HSL (100 nmol/l insulin, 0.49 +/- 0.05; P < 0.05), while no significant reduction was observed at other doses. Higher doses of insulin stimulated both HSL (1,000 nmol/l insulin, 1.4 +/- 0.07; P < 0.01) and LPL (control 1.00 +/- 0.0; 1,000 nmol/l insulin, 2.66 +/- 0.27; P < 0.01) protein expression. Cotreatment with RSG induced an increased dose response to insulin for LPL and HSL (P < 0.05); RSG alone also increased LPL and HSL expression (P < 0.05). Insulin stimulated TNF-alpha secretion in a dose-dependent manner (P < 0.01); the addition of RSG (10(-8) mol/l) reduced TNF-alpha secretion (P < 0.05). In summary, chronic treatment of human adipocytes with insulin stimulates lipolysis and LPL protein expression. The addition of RSG reduced the lipolytic rate and TNF-alpha secretion. The increase in lipolysis is not explained by changes in HSL expression. These data, therefore, may explain in part why hyperinsulinemia coexists with increased circulating nonesterified free fatty acids and increased adiposity in obese and/or type 2 diabetic patients.
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Tejido Adiposo/metabolismo , Hipoglucemiantes/farmacología , Insulina/farmacología , Lipólisis/efectos de los fármacos , Tiazoles/farmacología , Tiazolidinedionas , Adipocitos/efectos de los fármacos , Adipocitos/enzimología , Adipocitos/metabolismo , Tejido Adiposo/efectos de los fármacos , Adulto , Relación Dosis-Respuesta a Droga , Sinergismo Farmacológico , Femenino , Glicerol/metabolismo , Humanos , Técnicas In Vitro , Lipoproteína Lipasa/biosíntesis , Lipoproteína Lipasa/metabolismo , Persona de Mediana Edad , Norepinefrina/farmacología , Rosiglitazona , Esterol Esterasa/biosíntesis , Esterol Esterasa/metabolismo , Simpatomiméticos/farmacología , Factor de Necrosis Tumoral alfa/metabolismoRESUMEN
OBJECTIVE: Central obesity and sub-clinical inflammation increase metabolic risk, this study examined the intracellular inflammatory pathways in adipose tissue (AT) that contribute to this risk. DESIGN AND METHODS: This study therefore addressed the influence of NFκB and JNK activation in human abdominal subcutaneous (AbdSc) and omental (Om) AT, the effect of adiposity, T2DM status and the role of TNFα in vitro, using molecular biology techniques. RESULTS: Our data showed NFκB activity is increased in Om AT versus AbdSc AT (P<0.01), which was reversed with respect to depot specific activation of JNK (P<0.01). However, T2DM status appeared to preferentially activate NFκB (P<0.001) over JNK. Furthermore, in vitro studies showed recombinant human (rh) TNFα treated AbdSc adipocytes increased NFκB activity over time (2-48 h, P<0.05) whilst JNK activity reduced (2 h, 4 h, P<0.05); inhibitor studies supported a preferential role for NFκB as a modulator of TNFα secretion. CONCLUSIONS: These studies suggest distinct changes in NFκB and JNK activation, dependent upon AT depot, adiposity and T2DM status, with in vitro use of rh TNFα leading to activation of NFκB. Consequently NFκB appears to play a central role in inflammatory mediated metabolic disease over JNK, highlighting NFκB as a potential key target for therapeutic intervention.
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Tejido Adiposo/metabolismo , Adiposidad/fisiología , Diabetes Mellitus Tipo 2/genética , FN-kappa B/fisiología , Paniculitis/genética , Factor de Necrosis Tumoral alfa/fisiología , Tejido Adiposo/inmunología , Tejido Adiposo/patología , Adulto , Anciano , Células Cultivadas , Diabetes Mellitus Tipo 2/complicaciones , Diabetes Mellitus Tipo 2/metabolismo , Diabetes Mellitus Tipo 2/patología , Femenino , Humanos , Inflamación/genética , Inflamación/metabolismo , Masculino , Persona de Mediana Edad , Proteína Quinasa 8 Activada por Mitógenos/genética , Proteína Quinasa 8 Activada por Mitógenos/metabolismo , Proteína Quinasa 9 Activada por Mitógenos/genética , Proteína Quinasa 9 Activada por Mitógenos/metabolismo , Paniculitis/metabolismo , Proteínas Recombinantes/farmacología , Factor de Necrosis Tumoral alfa/farmacologíaRESUMEN
BACKGROUND AND PURPOSE: Hyperglycemia induced endoplasmic reticulum (ER) stress in diabetic vascular cells is considered an increasingly important factor for the genesis and development of atherosclerosis and cardiovascular complications. This study investigated firstly, the effect of hyperglycemia in ER stress induction in Human Umbilical Vein Endothelial Cells (HUVECs) and secondly, the impact of Glucagon like petide-1 (GLP-1) analogue, Liraglutide, in reducing ER stress in HUVECs exposed to high glucose (HG). EXPERIMENTAL APPROACH: HUVECs were incubated for 12 hr in 5 mmol/L normal glucose (NG) or in 25 mmol/L (HG) glucose with or without different concentrations of Liraglutide (1 nM, 10 nM or 100 nM) and components of ER stress pathways studied, using western blotting, to assess their expression levels. KEY RESULTS: Our data confirmed that exposure of HUVECs to HG up-regulated both up- (Bip/Grp78, PERK and IRE1α) and downstream (Calnexin, PDI and Ero1-Lα) markers of ER stress compared with control. Furthermore, Liraglutide showed a dose dependent capacity in preventing the onset of ER stress in HUVECs, with a maximum activity at 100 nM. HG also upregulated proapoptotic PUMA protein levels compared to controls. Interestingly, Liraglutide also induced OPA1, a marker of mitochondrial fusion, in a dose dependent manner. CONCLUSIONS AND IMPLICATIONS: Liraglutide prevented the onset of ER stress in human endothelial cells exposed to HG. Our data suggest that Liraglutide may exert its effects by inducing mitochondrial fusion processes, thus preventing HG induced mitochondrial fragmentation and apoptosis in human endothelial cells.
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Estrés del Retículo Endoplásmico/efectos de los fármacos , Péptido 1 Similar al Glucagón/análogos & derivados , Glucosa/antagonistas & inhibidores , Células Endoteliales de la Vena Umbilical Humana/citología , Células Endoteliales de la Vena Umbilical Humana/efectos de los fármacos , Células Cultivadas , Relación Dosis-Respuesta a Droga , Chaperón BiP del Retículo Endoplásmico , GTP Fosfohidrolasas/metabolismo , Péptido 1 Similar al Glucagón/farmacología , Glucosa/farmacología , Células Endoteliales de la Vena Umbilical Humana/metabolismo , Humanos , LiraglutidaRESUMEN
BACKGROUND: Antiretroviral (ARV) treatment has been associated with abnormalities in lipid and mitochondrial metabolism. We compared patterns of gene expression in the subcutaneous adipose tissue (SAT) of HIV-positive subjects before and after 18-24 months of ARV therapy with HIV-negative controls. METHODS: HIV patients naive to ARV were randomized to receive zidovudine (AZT), lamivudine (3TC) with efavirenz (EFV) or tenofovir disoproxil fumarate (TDF) with emtricitabine (FTC) and EFV. Healthy controls (n=15) were matched for age, ethnicity and gender. Patients on a regimen containing abacavir (ABC), 3TC and EFV for 18-24 months were also tested. Genes involved in adipocyte glucocorticoid, lipid and mitochondrial metabolism, and adipocyte differentiation, were profiled with real-time PCR. RESULTS: AZT led to increased visceral adipose tissue (VAT; P=0.012) and VAT:SAT ratio (P=0.036), whereas TDF increased SAT (P=0.047) and peripheral fat/lean body mass ratio (P=0.017). HIV treatment-naive patients had lower plasma lipoprotein lipase (LPL) activity (P=0.0001) versus controls (remaining below controls after ARV; P=0.038-0.0001). The overall pattern of gene expression was similar across all treatment groups, being most marked with AZT and least with TDF. There was up-regulation of peroxisome proliferator-activated receptor-γ coactivator-1α, uncoupling protein-2 and hexose 6-phosphate dehydrogenase, and down-regulation of nuclear respiratory factor-1, cytochrome oxidase B, cytochrome c oxidase-4, uncoupling protein-3, 11ß-hydroxysteroid dehydrogenase type-1, glucocorticoid receptor-α, fatty acid synthase, fatty acid binding protein-4, LPL and hormone sensitive lipase (18-24 months post-treatment versus pretreatment levels and controls; P<0.05 to <0.0001). CONCLUSIONS: The decreased expression of genes involved in lipid and mitochondrial metabolism 18-24 months post-ARV treatment in SAT of HIV patients, in conjunction with the increase in uncoupling protein-2 and decrease in cytochrome oxidase B gene expression, provides evidence of mitochondrial dysfunction and a shift to anaerobic metabolism within SAT in EFV-containing ARV regimens.
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Tejido Adiposo/metabolismo , Fármacos Anti-VIH/efectos adversos , Benzoxazinas/efectos adversos , Infecciones por VIH/tratamiento farmacológico , Lipogénesis/efectos de los fármacos , Inhibidores de la Transcriptasa Inversa/efectos adversos , Adenina/efectos adversos , Adenina/análogos & derivados , Adenina/farmacología , Adenina/uso terapéutico , Tejido Adiposo/efectos de los fármacos , Adulto , Alquinos , Anaerobiosis , Fármacos Anti-VIH/farmacología , Fármacos Anti-VIH/uso terapéutico , Benzoxazinas/farmacología , Benzoxazinas/uso terapéutico , Ciclopropanos , Quimioterapia Combinada , Complejo IV de Transporte de Electrones/genética , Complejo IV de Transporte de Electrones/metabolismo , Femenino , Regulación de la Expresión Génica , Infecciones por VIH/complicaciones , VIH-1/efectos de los fármacos , Humanos , Canales Iónicos/genética , Canales Iónicos/metabolismo , Masculino , Persona de Mediana Edad , Proteínas Mitocondriales/genética , Proteínas Mitocondriales/metabolismo , Organofosfonatos/efectos adversos , Organofosfonatos/farmacología , Organofosfonatos/uso terapéutico , Inhibidores de la Transcriptasa Inversa/farmacología , Inhibidores de la Transcriptasa Inversa/uso terapéutico , Tenofovir , Resultado del Tratamiento , Proteína Desacopladora 2 , Zidovudina/efectos adversos , Zidovudina/farmacología , Zidovudina/uso terapéuticoRESUMEN
A post-prandial increase in saturated fatty acids (SFAs) and glucose (Glc) activates an inflammatory response, which may be prolonged following restoration of physiological SFAs and Glc levels--a finding referred to as 'metabolic memory'. This study examined chronic and oscillating SFAs and Glc on the inflammatory signalling pathway in human adipose tissue (AT) and adipocytes (Ads) and determined whether Ads are subject to "metabolic memory." Abdominal (Abd) subcutaneous (Sc) explants and Ads were treated with chronic low glucose (L-Glc): 5.6 mM and high glucose (H-Glc): 17.5 mM, with low (0.2 mM) and high (2 mM) SFA for 48 h. Abd Sc explants and Ads were also exposed to the aforementioned treatment regimen for 12-h periods, with alternating rest periods of 12 h in L-Glc. Chronic treatment with L-Glc and high SFAs, H-Glc and high SFAs up-regulated key factors of the nuclear factor-κB (NFκB) pathway in Abd Sc AT and Ads (TLR4, NFκB; P<.05), whilst down-regulating MyD88. Oscillating Glc and SFA concentrations increased TLR4, NFκB, IKKß (P<.05) in explants and Ads and up-regulated MyD88 expression (P<.05). Both tumor necrosis factor α and interleukin 6 (P<.05) secretion were markedly increased in chronically treated Abd Sc explants and Ads whilst, with oscillating treatments, a sustained inflammatory effect was noted in absence of treatment. Therefore, SFAs may act as key instigators of the inflammatory response in human AT via NFκB activation, which suggests that short-term exposure of cells to uncontrolled levels of SFAs and Glc leads to a longer-term inflammatory insult within the Ad, which may have important implications for patients with obesity and Type 2 diabetes.
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Tejido Adiposo/metabolismo , Ácidos Grasos/efectos adversos , Inflamación/metabolismo , Receptor Toll-Like 4/metabolismo , Adipocitos/efectos de los fármacos , Adipocitos/metabolismo , Tejido Adiposo/efectos de los fármacos , Adulto , Femenino , Glucosa/farmacología , Humanos , Quinasa I-kappa B/metabolismo , Técnicas In Vitro , Inflamación/inducido químicamente , Interleucina-6/metabolismo , Persona de Mediana Edad , Proteína Quinasa 8 Activada por Mitógenos/metabolismo , Proteína Quinasa 9 Activada por Mitógenos/metabolismo , Factor 88 de Diferenciación Mieloide/metabolismo , FN-kappa B/metabolismo , Transducción de Señal , Factor 6 Asociado a Receptor de TNF/metabolismo , Pruebas de Toxicidad Crónica , Factor de Necrosis Tumoral alfa/metabolismoRESUMEN
OBJECTIVE: To evaluate the changes in circulating endotoxin after a high-saturated fat meal to determine whether these effects depend on metabolic disease state. RESEARCH DESIGN AND METHODS: Subjects (n = 54) were given a high-fat meal (75 g fat, 5 g carbohydrate, 6 g protein) after an overnight fast (nonobese control [NOC]: age 39.9 ± 11.8 years [mean ± SD], BMI 24.9 ± 3.2 kg/m(2), n = 9; obese: age 43.8 ± 9.5 years, BMI 33.3 ± 2.5 kg/m(2), n = 15; impaired glucose tolerance [IGT]: age 41.7 ± 11.3 years, BMI 32.0 ± 4.5 kg/m(2), n = 12; type 2 diabetic: age 45.4 ± 10.1 years, BMI 30.3 ± 4.5 kg/m(2), n = 18). Blood was collected before (0 h) and after the meal (1-4 h) for analysis. RESULTS: Baseline endotoxin was significantly higher in the type 2 diabetic and IGT subjects than in NOC subjects, with baseline circulating endotoxin levels 60.6% higher in type 2 diabetic subjects than in NOC subjects (P < 0.05). Ingestion of a high-fat meal led to a significant rise in endotoxin levels in type 2 diabetic, IGT, and obese subjects over the 4-h time period (P < 0.05). These findings also showed that, at 4 h after a meal, type 2 diabetic subjects had higher circulating endotoxin levels (125.4%↑) than NOC subjects (P < 0.05). CONCLUSIONS: These studies have highlighted that exposure to a high-fat meal elevates circulating endotoxin irrespective of metabolic state, as early as 1 h after a meal. However, this increase is substantial in IGT and type 2 diabetic subjects, suggesting that metabolic endotoxinemia is exacerbated after high fat intake. In conclusion, our data suggest that, in a compromised metabolic state such as type 2 diabetes, a continual snacking routine will cumulatively promote their condition more rapidly than in other individuals because of the greater exposure to endotoxin.
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Diabetes Mellitus Tipo 2/sangre , Dieta Alta en Grasa/efectos adversos , Endotoxinas/sangre , Adulto , Femenino , Humanos , Masculino , Persona de Mediana Edad , Periodo PosprandialRESUMEN
Central adiposity is a significant determinant of obesity-related hypertension risk, which may arise due to the pathogenic inflammatory nature of the abdominal fat depot. However, the influence of pro-inflammatory adipokines on blood pressure in the obese hypertensive phenotype has not been well established in Saudi subjects. As such, our study investigated whether inflammatory factors may represent useful biomarkers to delineate hypertension risk in a Saudi cohort with and without hypertension and/or diabetes mellitus type 2 (DMT2). Subjects were subdivided into four groups: healthy lean controls (age: 47.9±5.1 yr; BMI: 22.9±2.1 Kg/m(2)), non-hypertensive obese (age: 46.1±5.0 yr; BMI: 33.7±4.2 Kg/m(2)), hypertensive obese (age: 48.6±6.1 yr; BMI: 36.5±7.7 Kg/m(2)) and hypertensive obese with DMT2 (age: 50.8±6.0 yr; BMI: 35.3±6.7 Kg/m(2)). Anthropometric data were collected from all subjects and fasting blood samples were utilized for biochemical analysis. Serum angiotensin II (ANG II) levels were elevated in hypertensive obese (p<0.05) and hypertensive obese with DMT2 (p<0.001) compared with normotensive controls. Systolic blood pressure was positively associated with BMI (p<0.001), glucose (p<0.001), insulin (p<0.05), HOMA-IR (p<0.001), leptin (p<0.01), TNF-α (p<0.001) and ANG II (p<0.05). Associations between ANG II and TNF-α with systolic blood pressure remained significant after controlling for BMI. Additionally CRP (p<0.05), leptin (p<0.001) and leptin/adiponectin ratio (p<0.001) were also significantly associated with the hypertension phenotype. In conclusion our data suggests that circulating pro-inflammatory adipokines, particularly ANG II and, TNF-α, represent important factors associated with a hypertension phenotype and may directly contribute to predicting and exacerbating hypertension risk.
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Angiotensina II/sangre , Índice de Masa Corporal , Complicaciones de la Diabetes , Hipertensión/sangre , Obesidad/sangre , Factor de Necrosis Tumoral alfa/sangre , Adulto , Presión Sanguínea , Femenino , Humanos , Hipertensión/complicaciones , Masculino , Persona de Mediana Edad , Obesidad/complicaciones , Factores de Riesgo , Arabia SauditaRESUMEN
South Asians have a higher risk of type 2 diabetes mellitus (T2DM) and cardiovascular disease (CVD) than white Caucasians, for a given BMI. Premature biological ageing, assessed by reduction in telomere length (TL), may be mediated by factors resulting from altered metabolic profiles associated with obesity. We hypothesise that ethnicity and metabolic status represent detrimental factors contributing to premature biological ageing. Therefore we assessed TL in two South Asian, age and BMI-matched cohorts [T2DM (n = 142) versus non-T2DM (n = 76)] to determine the effects of BMI, gender, lipid and CVD profile on biological ageing. Genomic DNA was obtained from the UKADS cohort; biochemical and anthropometric data was collected and TL was measured by quantitative real-time PCR. Our findings indicated a gender-specific effect with reduced TL in T2DM men compared with non-T2DM men (P = 0.006). Additionally, in T2DM men, TL was inversely correlated with triglycerides and total cholesterol (r = -0.419, P < 0.01; r = -0.443, P < 0.01). In summary, TL was reduced amongst South Asian T2DM men and correlated with triglycerides and total cholesterol. This study highlights enhanced biological ageing among South Asian, T2DM men, which appears to be tracked by changes in lipids and BMI, suggesting that raised lipids and BMI may directly contribute to premature ageing.
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Envejecimiento/genética , Pueblo Asiatico/genética , Colesterol/sangre , Diabetes Mellitus Tipo 2/genética , Triglicéridos/sangre , Envejecimiento/sangre , Envejecimiento/etnología , Glucemia/genética , Estudios de Cohortes , Diabetes Mellitus Tipo 2/sangre , Diabetes Mellitus Tipo 2/etnología , Predisposición Genética a la Enfermedad , Humanos , Insulina/sangre , Resistencia a la Insulina/genética , Masculino , Persona de Mediana Edad , Acortamiento del TelómeroRESUMEN
Visfatin has been proposed as an insulin-mimicking adipocytokine, predominantly secreted from adipose tissue and correlated with obesity. However, recent studies suggest visfatin may act as a proinflammatory cytokine. Our studies sought to determine the significance of this adipocytokine and its potential role in the pathogenesis of T2DM. Firstly, we examined the effects of diabetic status on circulating visfatin levels, and several other adipocytokines, demonstrating that diabetic status increased visfatin*, TNF-α*** and IL-6*** compared with non-diabetic subjects (*p<0.05, **p<0.01, ***p<0.001, respectively). We then assessed the effects of an insulin sensitizer, rosiglitazone (RSG), in treatment naïve T2DM subjects, on circulating visfatin levels. Our findings showed that visfatin was reduced post-RSG treatment [vs. pre-treatment (*p<0.05)] accompanied by a reduction in HOMA-IR**, thus implicating a role for insulin in visfatin regulation. Further studies addressed the intracellular mechanisms by which visfatin may be regulated, and may exert pro-inflammatory effects, in human abdominal subcutaneous (Abd Sc) adipocytes. Following insulin (Ins) and RSG treatment, our in vitro findings highlighted that insulin (100 nM), alone, upregulated visfatin protein expression whereas, in combination with RSG (10 nM), it reduced visfatin*, IKKß** and p-JNK1/2*. Furthermore, inhibition of JNK protein exacted a significant reduction in visfatin expression (**p<0.01), whilst NF-κB blockade increased visfatin (*p<0.05), thus identifying JNK as the more influential factor in visfatin regulation. Additional in vitro analysis on adipokines regulating visfatin showed that only Abd Sc adipocytes treated with recombinant human (rh)IL-6 increased visfatin protein (*p<0.05), whilst rh visfatin treatment, itself, had no influence on TNF-α, IL-6 or resistin secretion from Sc adipocytes. These data highlight visfatin's regulation by insulin and RSG, potentially acting through NF-κB and JNK mechanisms, with only rh IL-6 modestly affecting visfatin regulation. Taken together, these findings suggest that visfatin may represent a pro-inflammatory cytokine that is influenced by insulin/insulin sensitivity via the NF-κB and JNK pathways.
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Adipocitos/enzimología , Citocinas/metabolismo , Diabetes Mellitus Tipo 2/enzimología , Proteínas Quinasas JNK Activadas por Mitógenos/metabolismo , FN-kappa B/metabolismo , Nicotinamida Fosforribosiltransferasa/metabolismo , Grasa Subcutánea Abdominal/citología , Tiazolidinedionas/farmacología , Adipocitos/efectos de los fármacos , Adipocitos/metabolismo , Adipoquinas/metabolismo , Adiposidad/efectos de los fármacos , Adulto , Estudios de Casos y Controles , Separación Celular , Medios de Cultivo Condicionados/farmacología , Citocinas/sangre , Citocinas/genética , Diabetes Mellitus Tipo 2/sangre , Diabetes Mellitus Tipo 2/diagnóstico , Diabetes Mellitus Tipo 2/tratamiento farmacológico , Femenino , Regulación de la Expresión Génica/efectos de los fármacos , Humanos , Quinasa I-kappa B/metabolismo , Inmunohistoquímica , Insulina/farmacología , Proteínas Quinasas JNK Activadas por Mitógenos/antagonistas & inhibidores , Masculino , FN-kappa B/antagonistas & inhibidores , Nicotinamida Fosforribosiltransferasa/sangre , Nicotinamida Fosforribosiltransferasa/genética , Fosforilación/efectos de los fármacos , Proteínas Recombinantes/farmacología , Rosiglitazona , Tiazolidinedionas/uso terapéutico , Factor de Necrosis Tumoral alfa/farmacologíaRESUMEN
BACKGROUND: Emerging data indicate that gut-derived endotoxin may contribute to low-grade systemic inflammation in insulin resistant states. This study aimed to examine the importance of serum endotoxin and inflammatory markers in non-alcoholic fatty liver disease (NAFLD) patients, with and without type 2 diabetes mellitus (T2DM), and to explore the effect of treatment with a lipase inhibitor, Orlistat, on their inflammatory status. METHODS: Fasted serum from 155 patients with biopsy proven NAFLD and 23 control subjects were analysed for endotoxin, soluble CD14 (sCD14), soluble tumour necrosis factor receptor II (sTNFRII) and various metabolic parameters. A subgroup of NAFLD patients were re-assessed 6 and 12 months after treatment with diet alone (n = 6) or diet plus Orlistat (n = 8). RESULTS: Endotoxin levels were significantly higher in patients with NAFLD compared with controls (NAFLD: 10.6(7.8, 14.8) EU/mL; controls: 3.9(3.2, 5.2) EU/mL, p < 0.001); NAFLD alone produced comparable endotoxin levels to T2DM (NAFLD: T2DM: 10.6(5.6, 14.2) EU/mL; non-diabetic: 10.6(8.5, 15.2) EU/mL), whilst a significant correlation between insulin resistance and serum endotoxin was observed (r = 0.27, p = 0.008). Both sCD14 (p < 0.01) and sTNFRII (p < 0.001) increased with severity of fibrosis. A positive correlation was also noted between sTNFRII and sCD14 in the NAFLD subjects (r = 0.29, p = 0.004).Sub-cohort treatment with Orlistat in patients with NAFLD showed significant decreases in ALT (p = 0.006), weight (p = 0.005) and endotoxin (p = 0.004) compared with the NAFLD, non-Orlistat treated control cohort at 6 and 12 months post therapy, respectively. CONCLUSIONS: Endotoxin levels were considerably increased in NAFLD patients, with marked increases noted in early stage fibrosis compared with controls. These results suggest elevated endotoxin may serve as an early indicator of potential liver damage, perhaps negating the need for invasive liver biopsy. As endotoxin may promote insulin resistance and inflammation, interventions aimed at reducing endotoxin levels in NAFLD patients may prove beneficial in reducing inflammatory burden.
RESUMEN
BACKGROUND: Circulating endotoxin levels are associated with atherosclerosis. Moreover, ethnic differences in pro-inflammatory markers may be associated with ethnic differences in atherosclerotic and cardiovascular (CVD) and coronary heart disease (CHD) risk. OBJECTIVE AND METHODS: To investigate ethnic differences in circulating plasma endotoxin levels, its soluble receptor (sCD14), and high-sensitivity CRP (hs-CRP). 192 individuals, aged 40-59 years (61 white (30 women), 68 of African origin (33 women) and 63 South Asians (33 women)), free from coronary heart disease (CHD), stroke, CVD and diabetes were randomly selected from the UK 'Wandsworth Heart and Stroke Study'. RESULTS: Age-adjusted endotoxin levels were lower in women than in men (p=0.002) and were highest in South Asians (13.3EU/mL [95% CI 12.0-14.7]) and lowest in individuals of African origin (10.1EU/mL [9.1-11.1]) than in whites (p for linear trend <0.001). Endotoxin levels were positively associated with waist, waist-hip ratio, total cholesterol, serum triglycerides and serum insulin levels and negatively associated with serum HDL-cholesterol. Serum hs-CRP and plasma sCD14 varied by ethnic group (p<0.001) but was not associated with endotoxin. CONCLUSIONS: This study is the first to indicate a graded increase in endotoxin levels from black Africans to whites to South Asians, which is consistent with the ethnic difference in CHD risk. Whilst these findings support the concept that the innate immune system (IIS) may contribute significantly to the metabolic component underlying the development of CVD and CHD risk, further studies are required to see whether endotoxin levels are causally related to the development of CHD.