Project description:C57Bl6/J male mice were put on different diets at 5 weeks of age, with a standard diet (SD) or a High-Fat High-Sucrose Diet (HFHS) or a Choline-Deficient High-Fat Diet (CDHFD) during 6 months. Primary hepatocytes cultures from 3 different models were synchronized in the cell cycle. Transcriptomic analysis was perfomed at 48hours of culture when HFHS and CDHFD hepatocytes harbor replication stress.
Project description:Genome wide DNA methylation profiling of liver tissues from Pemt-/- and Pemt+/+ under HFHS diets. Pemt-/- and Pemt+/+ mice were fed HFHS from 5 to 25 weeks of age and liver tissues were obtained at 25 weeks. Bisulphite converted DNAs were enriched with EpiXplore Methylation DNA Enrichment Kit (Clontech). Genomic DNA libraries was prepared by TruSeq ChIP Sample Preparation Kit.
Project description:We used microarrays to detail the global programme of gene expression in the liver of Pemt-/- and Pemt+/+ under HFHS diets. Liver sample of Pemt-/- and Pemt+/+ mice under HFHS for 1, 2, 3, 4 weeks were used for RNA extraction and hybridization on Affymetrix microarrays. Genes in liver of Pemt-/- mice, successively suppressed compared with Pemt+/+ mice, were obtained.
Project description:In order to study the heart disorder that the long term, high energy diet caused, Bama miniature pigs were fed a high-fat, high-sucrose diet for 23 months. These pigs developed symptoms of metabolic syndrome and showed cardiac steatosis and hypertrophy with a greatly increased heart weight (1.82-fold, P<0.05) and heart volume (1.60-fold, P<0.05) compared with the control pigs. To understand the molecular mechanisms of cardiac steatosis and hypertrophy, nine pig heart cRNA samples were hybridized to porcine GeneChips. The control group consisted of 6 Bama pigs fed a control diet, and the HFHSD group comprised 6 pigs that were induced with a HFHS diet, which included 37% sucrose, 53% control diet and 10% pork lard. The pigs were fed twice every day and provided water ad libitum for 23 months. The pigs were fasted for 12 hours and euthanized with ketamine and xylazine. Pig hearts from the HFHSD group pigs (120, 126, 138, 140, 144, and 146) and three control group pigs (157, 159, and 161) were sampled and preserved in liquid nitrogen and then for RNA extraction and hybridization on Affymetrix microarrays.
Project description:There is growing evidence that energy metabolism and insulin action are regulated by mechanisms that follow a diurnal rhythm and it has been proposed that defects in Akt signalling are associated with the pathophysiology of metabolic disease. It is therefore important to investigate these parameters under physiology of the free-living state. We therefore examined the insulin action in muscle of chow or high fat, high sucrose diet-fed (HFHS) rats during the normal diurnal cycle. HFHS animals displayed hyperinsulinemia, however had reduced systemic glucose disposal and impaired muscle glucose uptake during the feeding period. Proteomics and phosphoproteomics was performed over the diurnal cycle in chow and HFHS rats.
Project description:We have employed whole genome microarray expression profiling in the liver of mice fed a high-fat, high-sucrose diet for 15 weeks to search genes with N-terminal signal sequences whose expression is altered concomitantly with changes in islet size. At 15 weeks, this analysis revealed a significant >4-fold upregulation in 118 genes and downregulation by >75% in 22 genes encoding putative secreted factors in liver of mice fed a HFHS, although few changes (<2-fold upregulated or >one-half downregulated) were observed at 10 weeks. Expression of four genes (NRG1, GPNMB, SERPINB1 and HGF) from this signature was quantified in the different RNA samples by real-time PCR, confirming low variability between mice.
Project description:Excessive energy intake in modern society has led to an epidemic surge in metabolic diseases, such as obesity and type 2 diabetes. These disorders pose profound threats to women’s reproductive health. However, the precise impact and underlying pathogenesis of energy excess on female reproduction remain unclear. In this study, we established an obese and hyperglycemic female mouse model induced by a high-fat and high-sucrose (HFHS) diet, mimicking the modern lifestyle of overnutrition. These mice displayed marked reproductive dysfunction, including elevated serum testosterone levels, irregular estrous cycles, and impaired folliculogenesis. Analysis of the ovarian metabolome of the HFHS mice revealed a pronounced disturbance in amino acid metabolism. Transcriptomic profiling of these ovaries identified dysregulated genes involved in steroid hormone biosynthesis and glucolipid metabolism. Further multi-omic analysis unveils aberrations in ovarian arginine biosynthesis metabolism induced by HFHS diet. Notably, comparisons between HFHS mice and a cohort of women with polycystic ovary syndrome (PCOS) identified analogous reproductive and metabolic signatures. Therefore, our results provide direct in vivo evidence for the detrimental effects of overnutrition on female reproduction and offer insights into the metabolic underpinnings of PCOS.