The Immunometabolic Roles of Various Fatty Acids in Macrophages and Lymphocytes.

Macrophages and lymphocytes demonstrate metabolic plasticity, which is dependent partly on their state of activation and partly on the availability of various energy yielding and biosynthetic substrates (fatty acids, glucose, and amino acids). These substrates are essential to fuel-based metabolic reprogramming that supports optimal immune function, including the inflammatory response. In this review, we will focus on metabolism in macrophages and lymphocytes and discuss the role of fatty acids in governing the phenotype, activation, and functional status of these important cells. We summarize the current understanding of the pathways of fatty acid metabolism and related mechanisms of action and also explore possible new perspectives in this exciting area of research.

Association Between Aerobic Exercise and Rosiglitazone Avoided the NAFLD and Liver Inflammation Exacerbated in PPAR-α Knockout Mice.

Nonalcoholic fatty liver disease (NAFLD) is one of the main liver diseases today, and may progress to steatohepatitis, cirrhosis, and hepatocellular carcinoma. Some studies have shown the beneficial effects of aerobic exercise on reversing NAFLD. To verify whether chronic aerobic exercise improves the insulin resistance, liver inflammation, and steatohepatitis caused by a high fat diet (HF) and whether PPARα is involved in these actions. C57BL6 wild type (WT) and PPAR-α knockout (KO) mice were fed with a standard diet (SD) or HF during 12 weeks; the HF mice were trained on a treadmill during the last 8 weeks. Serum glucose and insulin tolerances, serum levels of aspartate aminotransferase, hepatic content of triacylglycerol, cytokines, gene expression, and protein expression were evaluated in all animals. Chronic exposure to HF diet increased triacylglycerol accumulation in the liver, leading to NAFLD, increased aminotransferase in the serum, increased peripheral insulin resistance, and higher adiposity index. Exercise reduced all these parameters in both animal genotypes. The liver lipid accumulation was not associated with inflammation; trained KO mice, however, presented a huge inflammatory response that was probably caused by a decrease in PPAR-γ expression. We conclude that exercise improved the damage caused by a HF independently of PPARα, apparently by a peripheral fatty acid oxidation in the skeletal muscle. We also found that the absence of PPARα together with exercise leads to a decrease in PPAR-γ and a huge inflammatory response. J. Cell. Physiol. 232: 1008-1019, 2017. © 2016 Wiley Periodicals, Inc.

Effect of an acute moderate-exercise session on metabolic and inflammatory profile of PPAR-α knockout mice.

Peroxisome proliferator-activated receptors (PPARs) play a major role in metabolism and inflammatory control. Exercise can modulate PPAR expression in skeletal muscle, adipose tissue, and macrophages. Little is known about the effects of PPAR-α in metabolic profile and cytokine secretion after acute exercise in macrophages. In this context, the aim of this study was to understand the influence of PPAR-α on exercise-mediated immune metabolic parameters in peritoneal macrophages. Mice C57BL/6 (WT) and PPAR-α knockout (KO) were examined in non-exercising control (n = 4) or 24 hours after acute moderate exercise (n = 8). Metabolic parameters (glucose, non-esterified fatty acids, total cholesterol [TC], and triacylglycerol [TG]) were assessed in serum. Cytokine concentrations (IL-1ß, IL-6, IL-10, TNF-α, and MCP-1) were measured from peritoneal macrophages cultured or not with LPS (2.5 µg/mL) and Rosiglitazone (1 µM). Exercised KO mice exhibited low glucose concentration and higher TC and TG in serum. At baseline, no difference in cytokine production between the genotypes was observed. However, IL-1ß was significantly higher in KO mice after LPS stimulus. IL-6 and IL-1ß had increased concentrations in KO compared with WT, even after exercise. MCP-1 was not restored in exercised KO LPS group. Rosiglitazone was not able to reduce proinflammatory cytokine production in KO mice at baseline level or associated with exercise. Acute exercise did not alter mRNA expression in WT mice. CONCLUSION: PPAR-α seems to be needed for metabolic glucose homeostasis and anti-inflammatory effect of acute exercise. Its absence may induce over-expression of pro-inflammatory cytokines in LPS stimulus. Moreover, moderate exercise or PPAR-γ agonist did not reverse this response.

Palmitoleic acid reduces the inflammation in LPS-stimulated macrophages by inhibition of NFκB, independently of PPARs.

Palmitoleic acid (PM, 16:1n-7) has anti-inflammatory properties that could be linked to higher expression of PPARα, an inhibitor of NFκB. Macrophages play a major role in the pathogenesis of chronic inflammation, however, the effects of PM on macrophages are underexplored. Thus, we aimed to investigate the effects of PM in activated macrophages as well the role of PPARα. Primary macrophages were isolated from C57BL/6 wild type (WT) and PPARα knockout (KO) mice, cultured under standard conditions and exposed to lipopolysaccharides LPS (2.5 µg/ml) and PM 600 µmol/L conjugated with albumin for 24 hours. The stimulation with LPS increased the production of interleukin (IL)-6 and IL-1ß while PM decreased the production of IL-6 in WT macrophages. In KO macrophages, LPS increased the production of tumour necrosis factor (TNF)-α and IL-6 and PM decreased the production of TNFα. The expression of inflammatory markers such NFκB and IL1ß were increased by LPS and decreased by PM in both WT and KO macrophages. PM reduced the expression of MyD88 and caspase-1 in KO macrophages, and the expression of TLR4 and HIF-1α in both WT and KO macrophages, although LPS had no effect. CD86, an inflammatory macrophage marker, was reduced by PM independently of genotype. PM increased PPARγ and reduced PPARß gene expression in macrophages of both genotypes, and increased ACOX-1 expression in KO macrophages. In conclusion, PM promotes anti-inflammatory effects in macrophages exposed to LPS through inhibition of inflammasome pathway, which was independent of PPARα, PPARϒ and AMPK, thus the molecular mechanisms of anti-inflammatory response caused by PM is still unclear.

Macadamia oil supplementation attenuates inflammation and adipocyte hypertrophy in obese mice.

Excess of saturated fatty acids in the diet has been associated with obesity, leading to systemic disruption of insulin signaling, glucose intolerance, and inflammation. Macadamia oil administration has been shown to improve lipid profile in humans. We evaluated the effect of macadamia oil supplementation on insulin sensitivity, inflammation, lipid profile, and adipocyte size in high-fat diet (HF) induced obesity in mice. C57BL/6 male mice (8 weeks) were divided into four groups: (a) control diet (CD), (b) HF, (c) CD supplemented with macadamia oil by gavage at 2 g/Kg of body weight, three times per week, for 12 weeks (CD + MO), and (d) HF diet supplemented with macadamia oil (HF + MO). CD and HF mice were supplemented with water. HF mice showed hypercholesterolemia and decreased insulin sensitivity as also previously shown. HF induced inflammation in adipose tissue and peritoneal macrophages, as well as adipocyte hypertrophy. Macadamia oil supplementation attenuated hypertrophy of adipocytes and inflammation in the adipose tissue and macrophages.

The association between skipping breakfast and biochemical variables in sedentary obese children and adolescents.

OBJECTIVE: To investigate the relationship between skipping meals and biochemical variables in obese children and adolescents. STUDY DESIGN: The sample was composed of 174 obese children and adolescents, aged between 6 and 16 years (80 male and 94 female). Body composition was assessed by dual-energy x-ray absorptiometry, fasting blood glucose, and lipid profile were measured after 12 hours fasting. The frequency of skipping breakfast, lunch, or dinner was assessed through a face-to-face interview carried out with the parents. RESULTS: The prevalence of eating breakfast daily was low in boys (47.5%) and girls (44.7%). A higher frequency of eating breakfast was negatively correlated with glucose (r = -0.16; P = .026), triglycerides (r = -0.19; P = .011), and very low density lipoprotein cholesterol (r = -0.21; P = .005). In the multivariate model, the weekly frequency of eating breakfast remained negatively associated with glucose (ß = -0.975; P = .017), triglycerides (ß = -7.792; P = .017), and very low density lipoprotein cholesterol (ß = -1.870; P = .009) independent of age, sex, trunk fatness, and parents' education. CONCLUSION: Skipping meals, mainly breakfast, is associated with glucose and lipid levels in obese children and adolescents.

Resting heart rate as a predictor of metabolic dysfunctions in obese children and adolescents.

BACKGROUND: Recent studies have identified that a higher resting heart rate (RHR) is associated with elevated blood pressure, independent of body fatness, age and ethnicity. However, it is still unclear whether RHR can also be applied as a screening for other risk factors, such as hyperglycemia and dyslipidemia. Thus, the purpose of the presented study was to analyze the association between RHR, lipid profile and fasting glucose in obese children and adolescents. METHODS: The sample was composed of 180 obese children and adolescents, aged between 7-16 years. Whole-body and segmental body composition were estimated by Dual-energy X-ray absorptiometry. Resting heart rate (RHR) was measured by heart rate monitors. The fasting blood samples were analyzed for serum triglycerides, total cholesterol (TC), high-density lipoprotein cholesterol (HDL-C), low-density lipoprotein cholesterol (LDL-C), and glucose, using the colorimetric method. RESULTS: Fasting glucose, TC, triglycerides, HDL-C, LDL-C and RHR were similar in both genders. The group of obese subjects with a higher RHR presented, at a lower age, higher triglycerides and TC. There was a significant relationship between RHR, triglycerides and TC. In the multivariate model, triglycerides and TC maintained a significant relationship with RHR independent of age, gender, general and trunk adiposity. The ROC curve indicated that RHR has a high potential for screening elevated total cholesterol and triglycerides as well as dyslipidemia. CONCLUSION: Elevated RHR has the potential to identify subjects at an increased risk of atherosclerosis development.

Doxorubicin modulated clock genes and cytokines in macrophages extracted from tumor-bearing mice.

Circadian rhythm is essential for cellular regulation of physiological, metabolic, and immune functions. Perturbations of circadian rhythms have been correlated with increased susceptibility to cancer and poor prognosis in the cancer treatment. Our aim is to investigate the role of doxorubicin (DOX) treatment on clock genes expression and inflammation in intraperitoneal macrophages and the antitumoral response. METHODS: Macrophages were extracted from intraperitoneal cavity of mice without or with Lewis lung carcinoma (LLC) and treated with DOX totaling four groups (CTL, LLC, LLC+DOX and DOX) and analyzes of clock genes in six time points (ZT02, ZT06, ZT10, ZT14, ZT18 AND ZT22). Intraperitoneal macrophages cell culture was stimulated with LPS and DOX and clock genes and inflammatory profile were analyzed. In tumor were analyzed macrophages markers. RESULTS: The expression of F4/80 (ZT22) and CD11c (ZT06) tumor tissue was significantly differed between LLC and LCC+DOX groups. In the intraperitoneal macrophages, DOX increased Clock (ZT10), Rev-Erbα (ZT18 and ZT22) and Per2 expressions (ZT18); in the LLC+DOX group was increased Bmal1 (ZT10), Per2 (ZT18) and NF-kB (ZT22) expressions; IL-6 expression increased in the LCC group (ZT02). In intraperitoneal macrophages cell culture stimulated with DOX and LPS after 24 h decreased Clock and Per1. DOX causes depression after 6 and 24 h in TNF-α content and Per2 gene expression after 24 h IL-1ß expression was reduced also. CONCLUSION: DOX treatment in vivo disrupted cytokine and clock genes expression in intraperitoneal macrophages suppressing immune response. Moreover, macrophages cultured with DOX had decreased expression of LPS-stimulated inflammatory cytokines.

Tributyrin in Inflammation: Does White Adipose Tissue Affect Colorectal Cancer?

Colorectal cancer affects the large intestine, leading to loss of white adipose tissue (WAT) and alterations in adipokine secretion. Lower incidence of colorectal cancer is associated with increased fibre intake. Fructooligosaccharides (FOS) are fibres that increase production of butyrate by the intestinal microbiota. Tributyrin, a prodrug of butyric acid, exerts beneficial anti-inflammatory effects on colorectal cancer. Our aim was to characterise the effects of diets rich in FOS and tributyrin within the context of a colon carcinogenesis model, and characterise possible support of tumorigenesis by WAT. C57/BL6 male mice were divided into four groups: a control group (CT) fed with chow diet and three colon carcinogenesis-induced groups fed either with chow diet (CA), tributyrin-supplemented diet (BUT), or with FOS-supplemented diet. Colon carcinogenesis decreased adipose mass in subcutaneous, epididymal, and retroperitoneal tissues, while also reducing serum glucose and leptin concentrations. However, it did not alter the concentrations of adiponectin, interleukin (IL)-6, IL-10, and tumour necrosis factor alpha (TNF)-α in WAT. Additionally, the supplements did not revert the colon cancer affected parameters. The BUT group exhibited even higher glucose tolerance and levels of IL-6, VEGF, and TNF-α in WAT. To conclude our study, FOS and butyrate supplements were not beneficial. In addition, butyrate worsened adipose tissue inflammation.

Metformin Mitigates Fibrosis and Glucose Intolerance Induced by Doxorubicin in Subcutaneous Adipose Tissue.

Doxorubicin (DX) is a chemotherapeutic drug that is used in clinical practice that promotes deleterious side effects in non-tumor tissues such as adipose tissue. We showed that DX leads to extensive damage in adipose tissue via a disruption in 5'-adenosine monophosphate-activated protein kinase (AMPK) and PPAR-gamma signaling. Thus, we investigated whether co-treatment with the biguanide drug metformin (MET) could prevent the side effects of DX through the activation of AMPK in adipose tissue. The goal of the present study was to verify the effects of DX and adjuvant MET treatment in subcutaneous adipose tissue (SAT) and to determine whether MET could protect against chemotherapy-induced side effects. C57/BL6 mice received DX hydrochloride (2.5 mg/kg) intraperitoneally 2 times per week for 2 weeks (DX), concomitantly or not, with MET administration (300 mg/kg oral daily) (DX + MET). The control group (CTRL) was pair-fed according to the food consumption of the DX group. After euthanasia, adipose tissue fat pads were collected, and SAT was extracted so that adipocytes could be isolated. Glucose uptake was then measured, and histological, gene, and protein analyses were performed. One-way analysis of variance was also performed, and significance was set to 5%. DX reduced retroperitoneal fat mass and epididymal pads and decreased glycemia. In cultured primary subcutaneous adipocytes, mice in the DX group had lower glucose uptake when stimulated with insulin compared with mice in the CTRL group. Adipocytes in the DX group exhibited a reduced area, perimeter, and diameter; decreased adiponectin secretion; and decreased fatty acid synthase gene expression. SAT from MET-treated mice also showed a reduction in collagen deposition. Treatment with MET prevented fibrosis and restored glucose uptake in SAT after insulin stimulation, yet the drug was unable to prevent other side effects of DX such as tissue loss and inflammatory response.

The role of moderate-to-vigorous physical activity in mediating the relationship between central adiposity and immunometabolic profile in postmenopausal women.

OBJECTIVES: To analyze the role of moderate-to-vigorous physical activity (MVPA) in mediating the relationship between central adiposity and immune and metabolic profile in postmenopausal women. MATERIALS AND METHODS: Cross-sectional study comprising 49 postmenopausal women (aged 59.26 ± 8.32 years) without regular physical exercise practice. Body composition was measured by dual-energy X-ray absorptiometry. Fasting blood samples were collected for assessment of nonesterified fatty acids, tumor necrosis factor-α (TNF-α), interleukin-6 (IL-6), adiponectin, insulin and estimation of insulin resistance (HOMA-IR). Physical activity level was assessed with an accelerometer (Actigraph GTX3x) and reported as a percentage of time spent in sedentary behavior and MVPA. All analyses were performed using the software SPSS 17.0, with a significance level set at 5%. RESULTS: Sedentary women had a positive relationship between trunk fat and IL-6 (rho = 0.471; p = 0.020), and trunk fat and HOMA-IR (rho = 0.418; p = 0.042). Adiponectin and fat mass (%) were only positively correlated in physically active women (rho = 0.441; p = 0.027). Physically active women with normal trunk fat values presented a 14.7% lower chance of having increased HOMA-IR levels (ß [95%CI] = 0.147 [0.027; 0.811]). CONCLUSIONS: The practice of sufficient levels of MVPA was a protective factor against immunometabolic disorders in postmenopausal women.

Morphological and metabolic determinants of nonalcoholic fatty liver disease in obese youth: a pilot study.

BACKGROUND: Nonalcoholic fatty liver disease (NAFLD) related to obesity has been rising in the last decades, though the morphological and metabolic determinants are remain unclear in children. The aim of this study was to analyze the morphological determinants and metabolic abnormalities in obese children and adolescents, classified either as with (P-NAFLD) or without (N-NAFLD). The sample comprised 190 individuals, aged 6 to 16 years-old, assigned into one of 4 groups according to sex and presence or absence of NAFLD. Obesity was obtained according to body mass index (BMI) cut-points. Body composition variables was estimated by Dual-Energy X-ray Absorptiometry (DEXA). Total cholesterol (TC), triglycerides (TG), HDL-cholesterol (HDL-C), LDL-cholesterol (LDL-C), insulin, fasting glucose (FG) and blood pressure were also analyzed. The diagnosis of NAFLD, as well as the measurement of intra-abdominal fat tissue thickness (IAF) and subcutaneous abdominal fat tissue thickness (SCF), was carried-out by ultrasound. RESULTS: Males and females belonging to P-NAFLD group showed, respectively, higher TFM and IAF. When data were adjusted for sex, age and total fat mass, those in P-NAFLD showed statistically higher IAF, TFM and TG. CONCLUSION: Our study showed that obese youngsters who were assigned to P-NAFLD group were twice as likely to present higher concentration of triglycerides, higher levels of trunk fat, as well as intra-abdominal fat compared to their N-NAFLD counterparts even after adjustments for sex, age, pubertal stage and total body fat mass.

The role of moderate-to-vigorous physical activity in mediating the relationship between central adiposity and immunometabolic profile in postmenopausal women

ABSTRACT Objectives To analyze the role of moderate-to-vigorous physical activity (MVPA) in mediating the relationship between central adiposity and immune and metabolic profile in postmenopausal women. Materials and methods Cross-sectional study comprising 49 postmenopausal women (aged 59.26 ± 8.32 years) without regular physical exercise practice. Body composition was measured by dual-energy X-ray absorptiometry. Fasting blood samples were collected for assessment of nonesterified fatty acids, tumor necrosis factor-α (TNF-α), interleukin-6 (IL-6), adiponectin, insulin and estimation of insulin resistance (HOMA-IR). Physical activity level was assessed with an accelerometer (Actigraph GTX3x) and reported as a percentage of time spent in sedentary behavior and MVPA. All analyses were performed using the software SPSS 17.0, with a significance level set at 5%. Results Sedentary women had a positive relationship between trunk fat and IL-6 (rho = 0.471; p = 0.020), and trunk fat and HOMA-IR (rho = 0.418; p = 0.042). Adiponectin and fat mass (%) were only positively correlated in physically active women (rho = 0.441; p = 0.027). Physically active women with normal trunk fat values presented a 14.7% lower chance of having increased HOMA-IR levels (β [95%CI] = 0.147 [0.027; 0.811]). Conclusions The practice of sufficient levels of MVPA was a protective factor against immunometabolic disorders in postmenopausal women.

Efeito de um protocolo de treinamento concorrente sobre fatores de risco para o acúmulo de gordura hepática de adolescentes obesos / Effect of a concurrent training on risk factors for the accumulation of hepatic fat of obese adolescents

Objetivo: Avaliar o efeito de um protocolo de treinamento concorrente com duração de 16 semanas sobre fatores de risco para o acúmulo de gordura hepática de jovens obesos. Metodologia: A amostra foi formada por 38 indivíduos obesos de ambos os sexos e com idade entre 12 e 15 anos. A obesidade foi atestada pelo percentual de gordura corporal, o qual foi estimado pela absortometria radiológica de dupla energia (DEXA). Adicionalmente, a quantidade de gordura localizada no tronco (kg) foi estimada também. Antes e após a intervenção, os jovens foram submetidos a exames bioquímicos de sangue(perfil lipídico completo em jejum [mg/dL]) e a ultrassonografia do fígado (tamanho dos lobos direito [LDem cm] e esquerdo [LE em cm]). A Intervenção consistiu de treinamento concorrente (treino resistido [30 minutos] e aeróbio [30 minutos]) com três sessões semanais, totalizando 180 minutos por semana. A análise estatística foi composta pelo teste t de Student para dados pareados, utilizando o software SPSS (17.0), e significância estatística fixada em p<5%. Resultados: Após a intervenção, foram observadas melhoras significantes no percentual de gordura total (PRÉ: 45,1±5,3 e PÓS: 41,7±5,6; p= 0,001) ena região do tronco (PRÉ: 46,5±5,6 e PÓS: 42,9±6,3; p= 0,001). Para o perfil lipídico, houve redução no colesterol total (PRÉ: 164±34 e PÓS: 148±29; p= 0,001), triglicérides (PRÉ: 118±59 e PÓS: 104±53; p=0,002) e lipoproteínas de baixa densidade (PRÉ: 100±29 e PÓS: 85±26; p= 0,001), porém, não para as de alta densidade (p= 0,981). Tanto o LE (PRÉ: 8,8±1,4 e PÓS: 7,8±1,3; p= 0,001) como o LD (PRÉ:13,6±1,3 e PÓS: 12,9±1,1; p= 0,001) sofreram diminuição em suas proporções. Conclusão: Em jovens obesos, o treinamento concorrente foi eficiente no combate a alguns fatores de risco ao acúmulo de gordura no fígado, bem como, na redução da gordura em ambos os lobos do órgão.

Objective: To evaluate the effect of a protocol of concurrent training lasting 16 weeks on risk factors for the accumulation of hepatic fat in obese youth. Methods: 38 obese children and adolescents of both sexes, between 12 and 15 years old. The obesity was attested by the percentage of body fat, which was estimated by dual-energy X-ray absorptiometry (DEXA). Additionally, the amount of fat located in the trunk (kg) was estimated too. Before and after the intervention, the youths underwent biochemical blood tests (fasting complete lipid profile [mg / dL]) and ultrasonography of the liver (right size Wolves [LD cm] and left [LE in cm]). The intervention consisted of concurrent training (strength training [30 minutes] and endurance training [30 minutes]) with three sessions per week, totaling 180 minutes a week, for ten weeks. Statistical analysis was made by the test t of Student for paired data using SPSS software (17.0) and significance statistical fixed at p <5%. Results: After the intervention, significant improvements were observed in the percentage of total fat (PRE: 45.1 ± 5.3 and POST: 41.7 ± 5.6, p = 0.001) and in the trunk region (PRE: 46, 5 ± 5.6 and POST: 42.9 ± 6.3, p = 0.001). For lipid profile, reduction in total cholesterol (PRE: 164 ± 34 and POST: 148 ± 29, p = 0.001), triglycerides (PRE: 118 ± 59 and POST: 104 ± 53, p =0.002) and lipoproteins density (PRE: 100 ± 29 and POST: 85 ± 26, p = 0.001), but not for high-density (p= 0.981). Both the LE (PRE: 8.8 ± 1.4 and POST: 7.8 ± 1.3, p = 0.001) and LD (PRE: 13.6 ± 1.3 and POST:12.9 ± 1, 1, p = 0.001) experienced a decrease in its proportions. Conclusion: The concurrent training was effective in combating some risk factors to the accumulation of fat in the liver, as well as in reducingfat in both lobes of the organ in young obese.