High-fat High-sucrose Diet Research Articles

The Slc25a47 locus is a novel determinant of hepatic mitochondrial function implicated in liver fibrosis.

Transporters of the SLC25 mitochondrial carrier superfamily bridge cytoplasmic and mitochondrial metabolism by channeling metabolites across mitochondrial membranes and are pivotal for metabolic homeostasis. Despite their physiological relevance as gatekeepers of cellular metabolism, most of the SLC25 family members remain uncharacterized. We undertook a comprehensive tissue distribution analysis of all Slc25 family members across metabolic organs and identified SLC25A47 as a liver-specific mitochondrial carrier. We used a murine loss-of-function model to unravel the role of this transporter in mitochondrial and hepatic homeostasis. We performed extensive metabolic phenotyping and molecular characterization of newly generated Slc25a47hep-/- and Slc25a47-Fgf21hep-/- mice. Slc25a47hep-/- mice displayed a wide variety of metabolic abnormalities, as a result of sustained energy deficiency in the liver originating from impaired mitochondrial respiration. This mitochondrial phenotype was associated with an activation of the mitochondrial stress response (MSR) in the liver, and the development of fibrosis, which was exacerbated upon feeding a high-fat high-sucrose diet. The MSR induced the secretion of several mitokines, amongst which FGF21 played a preponderant role on systemic physiology. To dissect the FGF21-dependent and -independent physiological changes induced in Slc25a47hep-/- mice, we generated a double Slc25a47-Fgf21hep-/- mouse model and demonstrated that several aspects of the hypermetabolic state were driven by hepatic secretion of FGF21. On the other hand, the metabolic fuel inflexibility observed in Slc25a47hep-/- mice could not be rescued with the genetic removal of Fgf21. Collectively, our data place the Slc25a47 locus at the center of mitochondrial homeostasis, which upon dysfunction triggers robust liver-specific and systemic adaptive stress responses. The prominent role of the Slc25a47 locus in hepatic fibrosis identifies this carrier, or its transported metabolite, as a potential target for therapeutic intervention. Herein, we report the importance ofa locus containing a liver-specific gene coding for a mitochondrial transport protein called SLC25A47. Mitochondria are the powerhouses of cells. They are crucial for metabolism and energy generation. We show that mice with genetic disruption of the Slc25a47 locus cannot maintain mitochondrial homeostasis (balance), leading to wide-ranging problems in the liver that have far-reaching physiological consequences.

Cepharanthine action in preventing obesity and hyperlipidemia in rats on a high-fat high sucrose diet

BackgroundIt was demonstrated that cepharanthine (CEP), derived from Stephania cepharantha hayata, is a potent inhibitor of the ABCC10 transmembrane protein. It is approved to be a natural product or remedy. The present study focuses on investigating whether cepharanthine effectively reduces hyperlipidemia and obesity in an experimental hyperlipidemic rat model. MethodFour groups of Wistar rats were assigned randomly to the following groups: a high-fat high sucrose diet (HFHS), normal-fat diet (NFD), HFHS plus cepraranthine (10 mg/kg) (HFHS-C), and a HFHS diet with atorvastatin (HFHS-A). The responses of rats were observed on the basis of serum and hepatic biochemical parameters, food intake, and body weight after CEP treatment, and assessing the histopathological modifications by the optical microscope in the liver and its cells. ResultsSignificant improvement in the serum total cholesterol (TC), serum triglycerides (TG), and serum low-density lipoprotein (LDL) levels were observed following CEP treatment. We have also observed significant improvement in the structure of liver tissue and reduced-fat droplets in the cytoplasm. Moreover, CEP had a significant effect in preventing the gain in body weight of animals, and food intake was not significantly affected. ConclusionOur research results revealed that CEP significantly improved dyslipidemia and prevented the accumulation of fatty deposits in the rats' liver tissue fed an HFHS diet. In addition, CEP exerted an anti-obesity effect.

Abstract 109: Mitochondrial Akap1 Protein Is Required For Diet-induced Obesity, Glucose Dysregulation And Hypertension

Mitochondria are best known as the powerhouse of the cell playing a critical role in energy metabolism with important implications in the development of obesity, a major cause of type 2 diabetes and hypertension. A-kinase anchoring protein 1 (AKAP1) is a mitochondrial scaffold protein that promote protein kinase A (PKA)-mediated phosphorylation of Drp1(Ser637) by increasing the local concentration of PKA at the outer mitochondrial membrane. However, the role of AKAP1 in the regulation of body weight, glucose homeostasis and blood pressure is not known. We used AKAP1 deficient mice to understand the physiological significance of this protein. Male and female AKAP1 -/- and AKAP1 +/- mice fed normal chow exhibit normal body weight relative to littermate controls. In contrast, AKAP1 -/- and AKAP1 +/- mice fed high fat high/sucrose diet (HFHSD) display attenuated weight gain compared to controls (male: 39.5 + 1.7 and 42.5 + 1.6 vs 47.3 + 2.3g, and female: 29.7 + 1.3 and 29.2 + 1.8 vs 32.5 + 1.5g). This was associated with significant decreased in fat mass in AKAP1 -/- (male:16.2 + 0.9g and female: 8.7 + 1.1g) and AKAP1 +/- (male:15.0 + 2.5g and female: 9.2 + 1.0g) mice compared to controls (male: 21.2 + 1.7g and female: 13.9 + 1.6g) whereas lean mass was not different between the three groups. Glucose tolerance test revealed that female AKAP1 -/- mice have improved glucose handling, and insulin tolerance test showed that insulin sensitivity is better in male AKAP1 -/- mice than controls. Notably, blood pressure was significantly lower in HFHSD-fed male AKAP1 -/- (systolic: 124.4 + 6 mmHg) and AKAP1 +/- (116.4 + 3 mmHg) mice vs control mice (146.4 + 5 mmHg). These findings demonstrated the importance of AKAP1 in the development of obesity and associated diabetes and hypertension. Our data also point to mitochondria function as a potential therapeutic target for treatment of common obesity and related diseases.

The Potential of Sorafenib in Preventing Metabolic Syndrome in Rats Fed a High-Fat High-Sucrose Diet

The purpose of this study was to investigate the possible preventive role of sorafenib in preventing obesity, hyperlipidemia, and diabetes by exploring its preventive effects. As there is a rapid increase in the number of people being affected by these diseases, there is a need to find new methods to manage them. Sorafenib has been demonstrated to be a potent inhibitor of ABCC10 transmembrane protein in several studies. A recent study has shown that ABCC10 contributes to the pathogenesis of both hyperglycemia and hyperlipidemia. Additionally, many patients who have cancer also are affected by the metabolic syndrome. Thus, cancer patients must take medicines to control high cholesterol and glucose levels, thus increasing the burden of medicines on cancer patients. As sorafenib is an anti-cancer drug and a potent inhibitor of ABCC10, it may prevent metabolic syndrome in cancer patients, thus reducing the burden of additional medications and their adverse effects. This study’s objective was to determine whether sorafenib can lower high lipid and high glucose levels in rats given a high-fat high-sugar diet. There were four groups of the rats: Group I: control (Standard Diet); Group II: diabetic (type-2) rats were given high-fat diet feed and sucrose through drinking water (25% sucrose) for 60 days (HFSD); Group III: diabetic (type-2) rats were given sorafenib 10 mg/kg/d (orally) for 60 days (HFS-S); Group IV: diabetic (type-2) rats were given metformin (50 mg/kg/d). Blood glucose, insulin, triglyceride, cholesterol, and liver enzyme levels were measured. Histopathological analysis of the liver was also conducted using an optical microscope. There was a significant weight reduction when sorafenib was administered to rats. The treatment produces a significant improvement in triglycerides (TG), total cholesterol (TC), and low-density lipoproteins (LDL) (P < 0.05). Furthermore, it also lowers blood glucose levels and improves insulin sensitivity (P < 0.05). Moreover, hepatic steatosis was also prevented by sorafenib in the histopathological analysis of the liver. According to our study, the effects of sorafenib were significant in improving dyslipidemia, hyperglycemia, and insulin sensitivity, as well as preventing the accumulation of fatty deposits in the rats' liver tissue when sorafenib was administered with a high-fat sucrose diet.

Exercise performed during pregnancy positively modulates liver metabolism and promotes mitochondrial biogenesis of female offspring in a rat model of diet-induced gestational diabetes

Gestational diabetes mellitus (GDM) is associated with a high-risk for metabolic complications in offspring. However, exercise is recognized as a non-pharmacological strategy against metabolic disorders and is recommended in GDM treatment. This study aimed to investigate whether gestational exercise (GE) could modulate maternal high-fat high-sucrose (HFHS) diet-related hepatic metabolic and mitochondrial outcomes in female offspring of mothers with HFHS-induced GDM.Female Sprague-Dawley rats were fed with control or HFHS diet and kept sedentary or submitted to GE. Their female offspring were fed with control diet and kept sedentary. Hepatic lipid accumulation, lipid metabolism regulators, mitochondrial biogenesis and dynamics markers, and microRNAs associated to the regulation of these markers were evaluated.Female offspring of GDM mothers showed increased body weight at early age, whereas GE prevented this effect of maternal HFHS-feeding and reduced hepatic lipid accumulation. GE stimulated hepatic mRNA transcription and protein expression of mitochondrial biogenesis markers (peroxisome proliferator-activated receptor-gamma co-activator-1alpha and mitochondrial transcription factor A) and mRNA transcription of mitochondrial dynamics markers (mitofusin-1, mitofusin-2, and dynamin-related protein-1) that were altered by maternal GDM, while mitochondrial dynamics markers protein expression was not affected by maternal diet/GE except for optic atrophy-1. MicroRNAs associated with these processes (miR-122, miR-34a, miR-130b, miR-494), and the expression of auto/mitophagy- and apoptosis-related proteins were not substantially influenced by altered intrauterine environment.Our findings suggest that GE is an important regulator of the intrauterine environment positively affecting liver metabolism and promoting liver mitochondrial biogenesis in female offspring despite eventual effects of maternal HFHS-feeding and related GDM.

Altered macronutrient composition and genetics influence the complex transcriptional network associated with adiposity in the Collaborative Cross

BackgroundObesity is a serious disease with a complex etiology characterized by overaccumulation of adiposity resulting in detrimental health outcomes. Given the liver’s critical role in the biological processes that attenuate adiposity accumulation, elucidating the influence of genetics and dietary patterns on hepatic gene expression is fundamental for improving methods of obesity prevention and treatment. To determine how genetics and diet impact obesity development, mice from 22 strains of the genetically diverse recombinant inbred Collaborative Cross (CC) mouse panel were challenged to either a high-protein or high-fat high-sucrose diet, followed by extensive phenotyping and analysis of hepatic gene expression.ResultsOver 1000 genes differentially expressed by perturbed dietary macronutrient composition were enriched for biological processes related to metabolic pathways. Additionally, over 9000 genes were differentially expressed by strain and enriched for biological process involved in cell adhesion and signaling. Weighted gene co-expression network analysis identified multiple gene clusters (modules) associated with body fat % whose average expression levels were influenced by both dietary macronutrient composition and genetics. Each module was enriched for distinct types of biological functions.ConclusionsGenetic background affected hepatic gene expression in the CC overall, but diet macronutrient differences also altered expression of a specific subset of genes. Changes in macronutrient composition altered gene expression related to metabolic processes, while genetic background heavily influenced a broad range of cellular functions and processes irrespective of adiposity. Understanding the individual role of macronutrient composition, genetics, and their interaction is critical to developing therapeutic strategies and policy recommendations for precision nutrition.

Role of the major terpenes of Callistemon citrinus against the oxidative stress during a hypercaloric diet in rats

1,8-Cineole, limonene and α-terpineol are the major terpenes present in Callistemon citrinus. This study reports for the first time that terpenes attenuate the oxidative stress in rats fed with high-fat-sucrose diet (HFSD) via antioxidant and anti-inflammatory mechanisms. Thirty-six male Wistar rats were divided into six groups (n = 6). Control (fed standard food, HFSD (fed with 41.7% fat and 16.6% sucrose), HFSD + 1,8-cineole (0.88 mg/kg body weight), limonene (0.43 mg/kg body weight), α-terpineol (0.32 mg/kg body weight) and a mixture of the three terpenes, given daily by gavage for 15 weeks. Morphometric and biochemical parameters were taken. Paraoxonase (PON1), reduced glutathione (GSH), lipid peroxidation products malondialdehyde (MDA) and hydroxyalkenals (HNE), advanced oxidation protein products (AOPP) and pro-inflammatory cytokines were measured in liver homogenates. All terpenes showed a remarkable reduction in weight gain, fat deposition, serum glucose and, triacylglycerol levels. However, terpenes presented different effects on the hepatic cell and the oxidative biomarkers. Conversely, the three terpenes and the mixture showed the same positive effect on the tumor necrosis factor alpha (TNFα), interleukin-6 (IL-6), leptin and adiponectin levels. Finally, 1,8-cineole, limonene and α-terpineol demonstrate significant anti-inflammatory effects and differential effects on the oxidative stress, suggesting the importance of these terpenes in Callistemon citrinus activities.

Beneficial effects of probiotic supplementation on glucose and triglycerides in a mouse model of metabolic syndrome

The present study aimed to examine the effect of Lactobacillus rhamnosus Rosell 11 and Lactobacillus helveticus Rosell 52 on glucose (blood level and tolerance), lipids (cholesterol and triglycerides), transaminases (AST and ALT), ALP, urea, and creatinine, along with body weight, food intake, liquid consumption, and gross pathology in a mouse model of metabolic syndrome. Male C57BL/6J mice were fed a high-fat high-sucrose diet and treated by oral gavage with a probiotic mixture in three different concentrations (107, 108, and 109 CFU/mL) once daily for 2 months. Probiotic supplementation, particularly at a concentration of 109 CFU, significantly decreased blood glucose and serum triglyceride levels, improved glucose tolerance, and promoted body weight loss in mice fed a high-fat high-sucrose diet. According to the obtained results, probiotic supplementation is useful for controlling glucose and triglyceride levels and could be used as an adjunctive therapeutic approach in patients with metabolic syndrome.

Rats with high aerobic capacity display enhanced transcriptional adaptability and upregulation of bile acid metabolism in response to an acute high-fat diet.

Rats selectively bred for the high intrinsic aerobic capacity runner (HCR) or low aerobic capacity runner (LCR) show pronounced differences in susceptibility for high‐fat/high sucrose (HFHS) diet‐induced hepatic steatosis and insulin resistance, replicating the protective effect of high aerobic capacity in humans. We have previously shown multiple systemic differences in energy and substrate metabolism that impacts steatosis between HCR and LCR rats. This study aimed to investigate hepatic‐specific mechanisms of action via changes in gene transcription. Livers of HCR rats had a greater number of genes that significantly changed in response to 3‐day HFHS compared with LCR rats (171 vs. 75 genes: >1.5‐fold, p < 0.05). HCR and LCR rats displayed numerous baseline differences in gene expression while on a low‐fat control diet (CON). A 3‐day HFHS diet resulted in greater expression of genes involved in the conversion of excess acetyl‐CoA to cholesterol and bile acid (BA) synthesis compared with the CON diet in HCR, but not LCR rats. These results were associated with higher fecal BA loss and lower serum BA concentrations in HCR rats. Exercise studies in rats and mice also revealed higher hepatic expression of cholesterol and BA synthesis genes. Overall, these results suggest that high aerobic capacity and exercise are associated with upregulated BA synthesis paired with greater fecal excretion of cholesterol and BA, an effect that may play a role in protection against hepatic steatosis in rodents.

Microbial and Transcriptomic Profiling Reveals Diet-Related Alterations of Metabolism in Metabolic Disordered Mice.

Metabolic disorders are the prelude of metabolic diseases, which are mainly due to the high-energy intake and genetic contribution. High-fat diet (HFD) or high-sucrose diet is widely used for inducing metabolic disorders characterized by increased body weight, insulin resistance, hepatic steatosis, and alteration of gut microbiome. However, the triangle relationship among diets, gut microbiome, and host metabolism is poorly understood. In our study, we investigated the dynamic changes in gut microbiota, and host metabolism in mice that were fed with either chow diet, HFD, or chow diet with 30% sucrose in drinking water (HSD) for continued 12 weeks. The gut microbiota was analyzed with 16S rDNA sequencing on feces. Hepatic gene expression profile was tested with transcriptomics analysis on liver tissue. The host metabolism was evaluated by measuring body weight, insulin sensitivity, serum lipids, and expression of proteins involved in lipid metabolism of liver. The results showed that HFD feeding affected body weight, insulin resistance, and hepatic steatosis more significantly than HSD feeding. 16S rRNA gene sequencing showed that HFD rapidly and steadily suppressed species richness, altered microbiota structure and function, and increased the abundance of bacteria responsible for fatty acid metabolism and inflammatory signaling. In contrast, HSD had minor impact on the overall bacteria structure or function but activated microbial bile acid biosynthesis. Fecal microbiota transplantation suggested that some metabolic changes induced by HFD or HSD feeding were transferrable, especially in the weight of white adipose tissue and hepatic triglyceride level that were consistent with the phenotypes in donor mice. Moreover, transcriptomic results showed that HFD feeding significantly inhibited fatty acid degradation and increase inflammation, while HSD increased hepatic de novo lipogenesis and inhibited primary bile acid synthesis alternative pathway. In general, our study revealed the dynamic and diversified impacts of HFD and HSD on gut microbiota and host metabolism.

Wheat alkylresorcinol increases fecal lipid excretion and suppresses feed efficiency in mice depending on time of supplementation

ObjectiveThe regular consumption of whole grains is linked to a lower likelihood of developing metabolic disorders. We previously found that chronic supplementation with wheat alkylresorcinols (ARs) prevents obesity and its associated metabolic symptoms induced by a high-fat high-sucrose diet (HFHSD) in mice. The aim of this study was to examine the time-of-day-dependence of these effects in mice. MethodsEight-wk-old male C57 BL/6 J mice were individually housed under a 12-h light/dark cycle (Zeitgeber time; ZT0, lights on; ZT12, lights off) and given access to a HFHSD from ZT12–16 (activity onset) and ZT20–24 (activity offset) to respectively represent breakfast and dinner times for 3 wk. Thereafter, the HFHSD was replaced with the same diet containing 0.4% ARs at either ZT12–16 or ZT20–24 for 8 wk. Control mice received the HFHSD without ARs at both feeding times. ResultsSupplementation with ARs significantly suppressed feed efficiency when given at breakfast, but not at dinner. ARs consumed at breakfast increased fecal lipid content and decreased the expression of Fat/Cd36 in enterocytes that enhances lipid uptake, but did not affect hepatic and blood lipid levels. The consumption of ARs at breakfast also upregulated the expression of Irs1, a key gene for insulin signaling in white adipose tissue and attenuated elevated blood leptin levels induced by the diet. This led to high scores for the homeostasis model assessment of insulin sensitivity, and the adiponectin/leptin ratio, a negative index of adipose tissue dysfunction. ConclusionsThese findings suggested that ARs ameliorate feed efficiency by decreasing dietary lipid absorption more effectively at the time of activity onset than offset. Further studies are needed to elucidate the molecular mechanism of the time-of-day-dependent effects of ARs on diet-induced metabolic disorders.

The effect of high-intensity interval training on serum and adipose tissues vaspin levels in rats fed a high-fat high-sucrose diet.

Vaspin is an adipocytokine with insulin-sensitizing and anti-inflammatory traits. The purpose of this study was to evaluate the effect of high-intensity interval training (HIIT) on serum, visceral and subcutaneous adipose tissue vaspin levels in rats exposed to a diet high in fat and sugar (HFS). Thirty-two male Wistar rats were randomly divided into HFS and standard diet (SD) groups. After 12weeks, each group was divided into sedentary and HIIT groups. HIIT program was performed 3times/week for 8weeks. Retroperitoneal adipose tissue, inguinal adipose tissue and serum were collected to analyze vaspin levels. Also, serum glucose and insulin levels, insulin resistance index (HOMA-IR) and retroperitoneal and inguinal fat weights were measured. HFS significantly increased weight gain, weight of inguinal (p=0.001) and retroperitoneal fat depots (p<0.001), serum glucose levels (p<0.001) and HOMA-IR (p<0.001). The HIIT was able to decline weight gain and fat mass (p<0.05) but did not affect inguinal and retroperitoneal fat depots' vaspin levels. Eight weeks' HIIT significantly increased serum vaspin (p=0.002) and decreased insulin (p=0.001) levels only in rats fed with SD. Although the HIIT program can cause significantly reducing effects on weight gain and fat depots' weights, it does not effect on circulating and fat depots' vaspin levels in rats fed an HFS.

10,12-Conjugated linoleic acid supplementation improves HDL composition and function in mice

Obesity is associated with inflammation, insulin resistance, and type 2 diabetes, which are major risk factors for CVD. One dietary component of ruminant animal foods, 10,12-conjugated linoleic acid (10,12 CLA), has been shown to promote weight loss in humans. Previous work has shown that 10,12 CLA is atheroprotective in mice by a mechanism that may be distinct from its weight loss effects, but this exact mechanism is unclear. To investigate this, we evaluated HDL composition and function in obese LDL receptor (Ldlr−/−) mice that were losing weight because of 10,12 CLA supplementation or caloric restriction (CR; weight-matched control group) and in an obese control group consuming a high-fat high-sucrose diet. We show that 10,12 CLA-HDL exerted a stronger anti-inflammatory effect than CR- or high-fat high-sucrose-HDL in cultured adipocytes. Furthermore, the 10,12 CLA-HDL particle (HDL-P) concentration was higher, attributed to more medium- and large-sized HDL-Ps. Passive cholesterol efflux capacity of 10,12 CLA-HDL was elevated, as was expression of HDL receptor scavenger receptor class B type 1 in the aortic arch. Murine macrophages treated with 10,12 CLA in vitro exhibited increased expression of cholesterol transporters Abca1 and Abcg1, suggesting increased cholesterol efflux potential of these cells. Finally, proteomics analysis revealed elevated Apoa1 content in 10,12 CLA-HDL-Ps, consistent with a higher particle concentration, and particles were also enriched with alpha-1-antitrypsin, an emerging anti-inflammatory and antiatherosclerotic HDL-associated protein. We conclude that 10,12 CLA may therefore exert its atheroprotective effects by increasing HDL-P concentration, HDL anti-inflammatory potential, and promoting beneficial effects on cholesterol efflux.

Effects of agavins in high fat-high sucrose diet-fed mice: an exploratory study

ABSTRACT The present study was to investigate whether agavins supplementation might reduce obesity in mice fed with a high fat-high sucrose (HF-HS) diet. Mice were fed with a HF-HS diet with (HF-HS+A) or without agavins (HF-HS) supplementation. Body weight, white adipose tissue (WAT), biomarkers inflammation, gastrointestinal hormones, microbiota, and their excreted metabolites were evaluated. HF-HS+A mice significantly reduced body weight, WAT, and leptin levels compared to the HF-HS group. Furthermore, pro-inflammatory cytokines and insulin levels tended to be lower in the HF-HS+A group. Moreover, the genera Allobaculum, Akkermansia, and Sutterella, linked with positives effects on host health, were identified as possible biomarkers for agavins treatment; meanwhile, ethyl oleate, thymine, hypoxanthine, uracil, and some fatty acids were substantially enriched with agavins and negative associated with pro-inflammatory biomarkers. Collectively, these results demonstrate that agavins can ameliorates many of the harmful effects induced by intake of a diet with a high fat and sucrose content.

Lipomodulatory and anti-oxidative stress effects of a polyherbal formulation based on garlic and avocado seed extracts on high fat high sucrose diet fed rats

ObjectiveTo determine antioxidant potentials of Allium sativum and Persea americana seeds extracts and three formulation-based extracts in vitro, and to evaluate the effects of the best formulation on oxidative stress and dyslipidemia on rats fed with high fat and high sucrose diet (HFHSD). MethodsAqueous extracts of Allium sativum, Persia. americana and three formulations were mixed at various portions (A. s/P. a; w/w): F (1:1), F (3: 1), and F(1:3). They were then tested for their antioxidant potentials in vitro using FRAP, DPPH and NO radicals to identify the best formulation. Four hundred (400) mg/kg b.w. of formulation F(1:1) were administered once daily for 21 days to rats previously fed with HFHSD for 8 weeks. Standard diet, vitamin E, and Atorvastatin were used as controls. After 21 days, body weight, blood glucose, lipid markers, activities of transaminases and markers of the antioxidant systems were assessed. ResultsThe Formulation F(1:1) showed the best in vitro activity with IC50 values of 6.5 and 2.23 mg/mL respectively for FRAP and DPPH- radical scavenging capacity. HFHSD caused a depletion of antioxidants associated with an increase of pro-oxidants and all the lipid markers except HDL-c Treatment with F(1:1) significantly increased TAC, SOD, and catalase activities, while MDA, protein carbonyls, and NO levels decreased (p < 0.05). Formulation F(1:1) decreased triglycerides (119.88 ± 4.25 mg/dL) and LDL-c (3.78 ± 0.66 mg/dL) levels and significantly increased the HDL-c level: (108.07 ± 6.29 mg/mL). Furthermore, Formulation F(1:1) significantly caused weight loss (2.31%), reduced blood glucose levels (27.38%) and ALT activity. ConclusionThe formulation F(1:1) could be a good candidate for the prevention and treatment of oxidative stress, dyslipidemia and features of metabolic syndrome.

COMBINED EFFECT OF HORSE GRAM AND FENUGREEK SEED POWDERS IN HIGH FAT-HIGH SUCROSE DIET-INDUCED PRE-DIABETES IN RATS

Objective: The objective of the study was to investigate the combined effect of horse gram (HG) and fenugreek (FG) seed powders in high fat-high sucrose (HFHS) diet-induced pre-diabetes in rats. Methods: Wistar rats (140–200 g) of either sex were randomly divided into four groups (n=6). Group 1, normal control, received normal pellet diet; Group 2, pre-diabetic control, received HFHS diet; Group 3, prophylactic group, received HFHS diet along with HG (200 mg/kg) and FG (200 mg/kg) combination; and Group 4, therapeutic group, received HFHS diet for 9 weeks followed by HG (200 mg/kg) and FG (200 mg/kg) combination for 28 days. Fasting blood glucose, lipid profiles, and OGTT were carried out on the 9th week in Group 1, Group 2, and Group 3, and at the end of the study in Group 1, Group 2, and Group 4. Results: Treatment with HFHS diet for 9 weeks has significantly increased fasting blood glucose and lipid profiles in the pre-diabetic group when compared to the normal control group indicating induction of pre-diabetes in the pre-diabetic group. In the prophylactic group, horse gram and fenugreek are given simultaneously with HFHS has significantly decreased fasting blood glucose, lipid profiles when compared to the pre-diabetic group. In the therapeutic group, treatment was given after induction of pre-diabetes for 28 days. In the therapeutic group, there was a significant decrease in fasting blood glucose, lipid profiles compared to the pre-diabetic group. Conclusion: HFHS diet-induced pre-diabetes or insulin resistance is an effective model to study and evaluate various new therapeutic modalities. The findings of the present study suggest that consumption of FG and HG as food supplements could reduce the risk of getting diabetes and progression of pre-diabetes to diabetes.

NFB-02. Maternal high-fat diet exposure increases the proliferation and glial differentiation of the cells of origin for NF1-optic glioma

Some pediatric cancers arise in the context of tumor predisposition syndromes caused by germline mutations in genes that regulate cell growth. The most common of these syndromes, Neurofibromatosis type 1 (NF1), results in the development of low-grade tumors of the optic pathway (optic pathway gliomas; OPGs) in ~15% of affected children. However, risk assessment for each individual child is difficult, as it is currently challenging to predict which children with NF1 will develop OPG. Emerging human population-based evidence has raised the possibility that patient-specific risk factors, including infant birth weight, may modulate the risk of glioma development. High-infant birth weight can be modeled in mice by maternal exposure to a high-fat high-sucrose (HFHS) diet. To address the hypothesis that maternal high-fat diet exposure will increase the risk of OPG formation through intrinsic effects on the putative tumor cell of origin, we evaluated the effect of maternal exposure to a HFHS diet on the proliferation and differentiation of these cells (neural stem cells within the third ventricular zone; TVZ NSCs). Progeny from obese dams (MatOb) exposed to HFHS diet during and preceding gestation demonstrated increased proliferation and glial differentiation of TVZ NSCs in vivo. Similar results were observed with gestational HFHS diet exposure (GE) that did not result in maternal weight gain, suggesting these effects were related to maternal diet rather than weight. These effects were not observed in NSCs from other regions of the brain, supporting the regional specificity of the effect of diet on these tumor-initiating stem cells. Finally, we found that maternal HFHS diet resulted in elevated levels of leptin and insulin, both of which increase the proliferation of TVZ NSCs. Taken together, these findings provide early experimental evidence that maternal environmental exposure affects developing neural stem cells relevant to the risk of glioma formation.

Raspberry Protects Against High-Fat, High-Sucrose Diet-Induced Vascular Oxidative Stress via NRF2

ObjectivesElevated free fatty acids in obesity and high-fat diets can negatively impact artery function, increasing the risk of cardiovascular diseases. Raspberries (RB) are a concentrated source of polyphenols which can mediate cellular pathways in a protective manner. Thus, the objective of this study was to examine whether RB consumption could mitigate the detrimental effects of a high-fat high-sucrose (HFHS) diet in the aorta of mice in vivo and human aortic endothelial cells (HAECs) in vitro. MethodsEight-week-old male C57BL/6 mice were fed an AIN-93M control diet with or without 10% w/w freeze-dried RB supplementation for 4 weeks. At week 4, animals were randomized into three groups: control, HFHS, or RB + HFHS. After 24 weeks of dietary treatment, mice were sacrificed and aortas dissected for immunohistochemistry analysis. HAECs were cultured in endothelial cell growth medium and treated with 200 μg/ml of RB extract in starvation medium. After 2 h, cells were treated with 100 μM of palmitic acid (PA) for 6 h. Cells were then collected for protein expression analysis via western blot. Data were normalized to control. Statistical analysis was conducted with ANOVA followed by Dunnett’s multiple comparisons test. ResultsIn aorta, nitrotyrosine was increased in the HFHS group compared with control and RB + HFHS group. Additionally, inducible nitric oxide synthase was increased in both HFHS and RB + HFHS compared to control, but expression was not different between HFHS groups. In HAECs, PA alone had no effect on phosphorylation of endothelial nitric oxide synthase (p-eNOSSer1177) compared to control. However, p-eNOSSer1177 expression was significantly increased in RB + PA -treated cells compared to both control and PA alone. Nuclear factor erythroid 2–related factor 2 (NRF2) and heme oxygenase (HO)-1 expression was also significantly increased with RB + PA treatment compared to control and PA alone. ConclusionsRaspberry consumption may reduce vascular oxidative stress induced by a HFHS diet via reduced NO oxidation despite increased iNOS in vivo. Further in vivo investigation is needed to confirm whether these effects are mediated by increased NRF2 activity. Funding SourcesLewis College Foundation and Agriculture and Food Research Initiative (grant no. 2019–67,017-29,257/project accession no. 1,018,642) from the USDA National Institute of Food and Agriculture.

250-OR: Role of Ezh2 in Hepatic Inflammation and Developmental Programming of Liver Disease

Maternal obesity predisposes offspring to the development of fatty liver and nonalcoholic steatohepatitis. Using a well-established mouse model (C57BL6/J) of maternal high-fat, high sucrose diet (HFD) exposure, we characterized epigenomic alterations in adult offspring liver. ChiP-seq for six histone modifications were performed on offspring livers from dams fed either a AIN93G (CON, 7% fat calories, n=11) or HFD (45% fat calories, n=9) for 12 wk prior to pregnancy and through weaning. Offspring of HFD dams, showed a robust gain in repressive H3K27me3 modification mediated by the histone methyltransferase, Ezh2. Since Ezh2 has been implicated in modulating inflammatory signaling we examined if Ezh2 in macrophages contributes to hepatic inflammation following a high-fat palmitate/cholesterol (FPC) diet. We generated myeloid cell-specific Ezh2-knockouts (Ezh2-CKO) by crossing Ezh2-floxed (Ezh2f/f) mice to LysM-Cre expressing mice. At 5 weeks of age Ezh2-CKO and flox-control litter mates were provided ad libitum CON (n=22) or FPC diet (52% fat calories, n=22) for 14 days. At 7 weeks of age, body and liver weights were collected and liver tissue was flash frozen for RNA-seq analyses. Two-way ANOVA of liver to body weight ratios showed a significant diet effect (p&amp;lt;0.0001) , with increased ratios in the FPC group without effects of genotype. Transcriptomic analyses showed that the effects of FPC diet were diminished in CKO mice relative to flox controls (223 vs.132 DEGs in flox and CKO, respectively) with 2DEGs shared between genotypes. Ingenuity Pathway Analysis revealed signaling pathways and processes related to inflammation, macrophage and leukocyte activation, reactive oxygen species, and PPARα that were increased in the flox with FPC diets compared to CKO animals. Collectively, these findings reveal a role for Ezh2 in developmental programing of liver disease and hepatic inflammation, uncovering a novel epigenetic process influencing hepatic disease risk in offspring. Disclosure M.L.Ruebel: None. P.Jambal: None. S.V.Chintapalli: None. J.Chen: None. K.Shankar: None. Funding U.S. Department of Agriculture-ARS-CRIS (6251-51000-005-00D) NIH (P30DK048520-27, R01HD102726-01A1) Funds from the Department of Pediatrics, University of ColoradoAnschutz Medical Campus and the Anschutz Health and Wellness Center.

Metformin does not prevent the alteration of reticulum-mitochondria Ca2+ coupling and the progression towards early diabetic cardiomyopathy with HFpEF in a diet-induced mouse model of T2D

Diabetic cardiomyopathy, as a major complication of type 2 diabetes (T2D), shows a rising prevalence nowadays. Reticulum-mitochondria Ca2+ uncoupling was recently reported as an early but reversible trigger of cardiac dysfunction using a diet-induced murine model of DCM with heart failure and preserved ejection fraction (HFpEF). Metformin is the first-line treatment prescribed to treat T2D patients, with known protective effects against myocardial infarction. We here aimed to determine if metformin could hinder the alteration of the reticulum-mitochondria Ca2+ coupling and the cardiac dysfunction induced by a high-fat high-sucrose diet (HFHSD) in mice. Mice were either fed a HFHSD or a standard diet for 16 weeks. During the last 6 weeks of diet, HFHSD mice received a daily oral gavage with either metformin or vehicle. Reticulum-mitochondria Ca2+ coupling was assessed: 1) structrally by proximity ligation assay between IP3R and VDAC; and 2) functionally by Ca2+ imaging of the mitochondrial Ca2+ sensor, 4mtD3cpv, expressed by intramyocardial adenoviral injection. Echocardiography was perfomed on lightly anesthetized animals to assess systolic function and hypertrophy. Metformin rescued the glucose tolerance and insulinemia in HFHSD mice, but partially for the cardiac insulin resistance. However, in the HFHSD cardiomyocyte, metformin did not preclude the reduction of the proximity between IP3R and VDAC, and of the histamine-driven reticulum-mitochondria Ca2+ transfer through the IP3R-VDAC complex. Echocardiography also revealed no prevention of HFHSD-induced cardiac hypertrophy and strain rate reduction. Our data report that 6 weeks of metformin treatment did not preserve the reticulum-mitochondria Ca2+ coupling in the diabetic heart and did not prevent the progression towards early DCM with HFpEF.