Project description:Dietary consumption of long-chain omega-3 polyunsaturated fatty acids (n-3 PUFA) may protect against cardiometabolic disease through modulation of systemic and adipose inflammation. However, it is often difficult to detect the subtle effects of n-3 PUFA on inflammatory biomarkers in traditional intervention studies. We aimed to identify novel n-3 PUFA modulated gene expression using unbiased adipose transcriptomics during evoked endotoxemia in a clinical trial of n-3 PUFA supplementation. We analyzed adipose gene expression using RNA sequencing in the fenofibrate and omega-3 fatty acid modulation of endotoxemia (FFAME) trial of healthy individuals at three timepoints: before and after n-3 PUFA supplementation (n=8; 3600mg/day EPA/DHA) for 6weeks compared with placebo (n=6), as well as during a subsequent evoked inflammatory challenge (lipopolysaccharide 0.6ng/kg i.v.). As expected, supplementation with n-3 PUFA vs. placebo alone had only modest effects on adipose tissue gene expression. In contrast, the transcriptomic response to evoked endotoxemia was significantly modified by n-3 PUFA supplementation, with several genes demonstrating significant n-3 PUFA gene-nutrient interactions. These data highlight potential mechanisms whereby n-3 PUFA consumption may enhance the immune response to an inflammatory challenge.
Project description:Gene expresion profiles from the scAT following 6 week LC n-3 PUFA and 6 week placebo supplementation were compared Women with PCOS were supplemented with 4g n-3 PUFA (containing 1.8g EPA and DHA) daily for 6 weeks and changes in subcutaneous adipose tissue gene expression was compared with 6 week placebo supplementation.
Project description:Gene expresion profiles from the scAT following 6 week LC n-3 PUFA and 6 week placebo supplementation were compared Women with PCOS were supplemented with 4g n-3 PUFA (containing 1.8g EPA and DHA) daily for 6 weeks and changes in subcutaneous adipose tissue gene expression was compared with 6 week placebo supplementation. This was a cross-over placebo, controlled dietary intervention wherein women with PCOS received both treatments.
Project description:We report that the HF/HS-mediated functional enrichment of genes of immunity and inflammation is driven toward normal by the AOF supplementation Obesity may not constantly associate with metabolic disorders and mortality later in life, raising the challenging concept of healthy obesity. Here, high fat-high sucrose (HF/HS) feeding produces hyperglycaemia and hypercholesterolemia, increases oxidative stress, elevates endotoxemia, expands adipose tissue (with enlarged adipocytes, macrophage infiltration and accumulation of cholesterol and oxysterols), and reduces lifespan of obese mice. Despite persistence of obesity, supplementation with an antioxidant formulation normalizes plasma lipids and endotoxemia, prevents macrophage recruitment in adipose tissue, reduces adipose accumulation of cholesterol and cholesterol oxides, and extends lifespan. The HF/HS-mediated functional enrichment of genes of immunity and inflammation (in particular response to lipopolysaccharides) is driven towards normal by the antioxidant formulation. It is concluded that the limitation of immune cell infiltration in adipose tissue on the long term by an antioxidant formulation can increase lifespan independently of body weight and fat storage. It constitutes the hallmark of a healthy adiposity trait.
Project description:We assessed the effects of supplementing milk fat globules (MFG) on the growth and development of the skeleton in rats fed a Western unbalanced diet (UBD). The UBD is high in sugar and fat, low in protein, fiber, and micronutrients, and negatively impacts health. The MFG—a complex structure secreted in milk—has a unique proteome and lipidome, and differs significantly from isolated dietary ingredients. Rats consuming the UBD exhibited growth retardation and disrupted bone structural and mechanical parameters; these were improved by supplementation with small MFG. The addition of small MFG increased the efficiency of protein utilization for growth, and improved trabecular and cortical bone parameters. Furthermore, consumption of UBD led to a decreased concentration of saturated fatty acids and increased levels of polyunsaturated fatty acids (PUFA), particularly omega-6 PUFA, in the serum, liver, and adipose tissue. The addition of small MFG restored PUFA concentration and the ratio of omega-6 to omega-3 PUFA in bone marrow and adipose tissue. Finally, large but not small MFG supplementation affected the cecal microbiome in rats. Overall, our results suggest that natural structure MFG supplementation can improve metabolism and bone development in rats fed an UBD, with the effects depending on MFG size, whereas the detrimental effects of an UBD on the microbiome were not mitigated by MFG supplementation.
Project description:Inappropriate or sustained activation of innate immunity is a pathologic feature of several common cardio-metabolic disorders. Little is known, however, about transcriptomic modulation during inflammatory stress in disease-relevant human tissues. We applied deep RNA sequencing (RNA-seq) during low-dose experimental endotoxemia (LPS) in healthy humans to interrogate, in an unbiased manner, inflammatory tissue-level transcriptome responses of relevance to complex cardio-metabolic diseases. We utilized adipose and blood samples from three individuals who underwent a standardized inpatient endotoxemia protocol. Our comprehensive analysis revealed substantial, highly tissue- and subject-specific LPS-modulated changes in the expression of protein-coding genes and linc-RNAs as well as alternative splicing (AS). We also confirmed adipocytes and macrophages as potential cell sources of selective LPS-modulated linc-RNAs and AS events. Finally, we defined disease relevance of a subset of findings in obese adipose tissue and through interrogation of overlap with genome-wide association study loci for cardio-metabolic traits. Our findings provide novel insights into tissue-level genomic regulation, not detectable through analysis of DNA variations alone, of relevance to common cardio-metabolic diseases. Using RNA-seq data to study LPS-modulated changes in lincRNA expression for adipose and blood of a healthy individual.
Project description:The objectives of this study were to examine the effect of (i) dietary supplementation with n-3 PUFA and (ii) post-insemination plane of nutrition on the global transcriptome of uterine endometrial tissue, with particular focus on key genes and pathways that may affect pregnancy outcome. A total of 60 estrous-synchronised continental crossbred heifers were offered a concentrate-based diet supplemented (n-3 PUFA; n=32) or not (Control; n=28) with an n-3 PUFA-enriched product for 30 days prior to insemination. Following insemination, animals were allocated, within treatment, to one of two post insemination diets, either remaining on their existing high plane of nutrition (Control + High (n = 14); n-3 PUFA + High (n = 15)), or fed a low plane of nutrition diet (Control + Low (n = 14); n-3 PUFA + Low (n = 17)). Uterine endometrial tissue was collected, RNA isolated and gene expression analysis conducted using RNAseq. Comparison of transcript abundance across groups, highlighted a significant effect of diet on the endometrial transcriptome. The mTOR signalling pathway as well as pathways involved in early pregnancy/fertility was significantly enriched as a consequence of diet.
Project description:Omega-3 polyunsaturated fatty acids (n-3 PUFA), such as the eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA), are reported to beneficially affect intestinal immunity. The intestinal epithelial layer (IEL), made up of absorptive cells as the enterocytes, is the primary site of fatty acid absorption. So far, the biological pathways modulated by n-3 PUFA in IEL during infection remain elusive. In the present study, shotgun proteomics analysis integrated with RNA sequencing (RNA-seq) technology was employed to unveil the proteomic changes induced by n-3 PUFA in enterocytes, in the presence and absence of lipopolysaccharide (LPS) stress conditions. A total of 33, 85, and 88 differential proteins were identified in the IPEC-J2 cells exposed to (i) n-3 PUFA (DHA:EPA, 1:2, 10 µM, 24 h), (ii) LPS (10 µg/mL, 24 h), or (iii) n-3 PUFA pre-treatment followed by LPS stimulation, respectively. Functional annotation and pathway analysis of the differential proteins revealed the modulation of central carbon metabolism, including glycolysis/gluconeogenesis, pentose phosphate pathway, and oxidative phosphorylation. Specifically, LPS caused metabolic dysregulation in the IPEC-J2 cells, abated upon prior treatment with n-3 PUFA. Indeed, n-3 PUFA supplementation facilitated enterocyte development and lipid homeostasis. This work provided a comprehensive picture of the biological pathways modulated by n-3 PUFA in enterocytes, both in the absence and presence of a state of endotoxin-induced metabolic dysregulation. Our findings provide important insights into the role of n-3 PUFA in the intestinal barrier, which could support better planning of nutritional strategies in managing intestinal health and immunity.
Project description:Inappropriate or sustained activation of innate immunity is a pathologic feature of several common cardio-metabolic disorders. Little is known, however, about transcriptomic modulation during inflammatory stress in disease-relevant human tissues. We applied deep RNA sequencing (RNA-seq) during low-dose experimental endotoxemia (LPS) in healthy humans to interrogate, in an unbiased manner, inflammatory tissue-level transcriptome responses of relevance to complex cardio-metabolic diseases. We utilized adipose and blood samples from three individuals who underwent a standardized inpatient endotoxemia protocol. Our comprehensive analysis revealed substantial, highly tissue- and subject-specific LPS-modulated changes in the expression of protein-coding genes and linc-RNAs as well as alternative splicing (AS). We also confirmed adipocytes and macrophages as potential cell sources of selective LPS-modulated linc-RNAs and AS events. Finally, we defined disease relevance of a subset of findings in obese adipose tissue and through interrogation of overlap with genome-wide association study loci for cardio-metabolic traits. Our findings provide novel insights into tissue-level genomic regulation, not detectable through analysis of DNA variations alone, of relevance to common cardio-metabolic diseases.
Project description:The purpose of this study was to evaluate the effect of endotoxemia on brown adipose tissue by injection of ultrasonicated P. gingivalis to mice