High-fat High-sugar Diet Research Articles

Gamma-oryzanol as a potential modulator of oxidative stress and inflammation via PPAR-y in adipose tissue: a hypothetical therapeutic for cytokine storm in COVID-19?

The literature has reported a higher prevalence of negative clinical outcomes due to Coronavirus disease 19 (COVID-19) in obese individuals. This can be explained by the cytokine storm, result from the cytokine production from both obesity and viral infection. Gamma-oryzanol (γOz) is a compound with anti-inflammatory and antioxidant activities. However, little is known about the γOz action as a possible agonist of peroxisome proliferator-activated receptor gamma (PPAR-γ). The aim of this study was to test the hypothesis that γOz attenuates the cytokine storm by stimulating PPAR-γ in the adipose tissue. Methods: Male Wistar rats were randomly divided into three experimental groups and fed ad libitum for 30 weeks with control diet (C, n = 6), high sugar-fat diet (HSF, n = 6) or high sugar-fat diet + γOz (HSF + γOz, n = 6). HSF groups also received water + sucrose (25%). The γOz dose was 0.5% in the chow. Evaluation in animals included caloric intake, body weight, adiposity index, plasma triglycerides, and HOMA-IR. In adipose tissue was evaluated: PPAR-γ gene and protein expression, inflammatory and oxidative stress parameters, and histological analysis. Results: Adipose tissue dysfunction was observed in HSF group, which presented remarkable PPAR-γ underexpression and increased levels of cytokines, other inflammatory markers and oxidative stress. The γOz treatment prevented adipose tissue dysfunction and promoted PPAR-γ overexpression. Conclusion: Natural compounds as γOz can be considered a coadjutant therapy to prevent the cytokine storm in COVID-19 patients with obesity conditions.

Metaplasticity in the Ventral Pallidum as a Potential Marker for the Propensity to Gain Weight in Chronic High-Calorie Diet.

A major driver of obesity is the increasing palatability of processed foods. Although reward circuits promote the consumption of palatable food, their involvement in obesity remains unclear. The ventral pallidum (VP) is a key hub in the reward system that encodes the hedonic aspects of palatable food consumption and participates in various proposed feeding circuits. However, there is still no evidence for its involvement in developing diet-induced obesity. Here we examine, using male C57BL6/J mice and patch-clamp electrophysiology, how chronic high-fat high-sugar (HFHS) diet changes the physiology of the VP and whether mice that gain the most weight differ in their VP physiology from others. We found that 10-12 weeks of HFHS diet hyperpolarized and decreased the firing rate of VP neurons without a major change in synaptic inhibitory input. Within the HFHS group, the top 33% weight gainers (WGs) had a more hyperpolarized VP with longer latency to fire action potentials on depolarization compared with bottom 33% of weight gainers (i.e., non-weight gainers). WGs also showed synaptic potentiation of inhibitory inputs both at the millisecond and minute ranges. Moreover, we found that the tendency to potentiate the inhibitory inputs to the VP might exist in overeating mice even before exposure to HFHS, thus making it a potential property of being an overeater. These data point to the VP as a critical player in obesity and suggest that hyperpolarized membrane potential of, and potentiated inhibitory inputs to, VP neurons may play a significant role in promoting the overeating of palatable food.SIGNIFICANCE STATEMENT In modern world, where highly palatable food is readily available, overeating is often driven by motivational, rather than metabolic, needs. It is thus conceivable that reward circuits differ between obese and normal-weight individuals. But is such difference, if it exists, innate or does it develop with overeating? Here we reveal synaptic properties in the ventral pallidum, a central hub of reward circuits, that differ between mice that gain the most and the least weight when given unlimited access to highly palatable food. We show that these synaptic differences also exist without exposure to palatable food, potentially making them innate properties that render some more susceptible than others to overeat. Thus, the propensity to overeat may have a strong innate component embedded in reward circuits.

Exercise and/or Genistein Treatment Impact Gut Microbiota and Inflammation after 12 Weeks on a High-Fat, High-Sugar Diet in C57BL/6 Mice.

Genistein (Gen) and exercise (Exe) have been postulated as potential strategies to ameliorate obesity, inflammation, and gut microbiota (GM) with promising results. However, the impact of the combination of both Exe and Gen is yet to be investigated. We aimed to analyze the impacts of Exe, Gen, and their combined effects on GM and inflammation in mice after a 12-week high-fat, high-sugar diet (HFD). Eighty-three C57BL/6 mice were randomized to control, HFD, HFD + Exe, HFD + Gen, or HFD + Exe + Gen. The V4 region of the 16S rRNA gene was analyzed with Illumina MiSeq. Serum samples were used to analyze interleukin (Il)-6 and Tumor Necrosis Factor alpha (TNF-alpha). The HFD + Exe and HFD + Exe + Gen treatments resulted in significantly greater microbial richness compared to HFD. All the treatments had a significantly different impact on the GM community structure. Ruminococcus was significantly more abundant after the HFD + Exe + Gen treatment when compared to all the other HFD groups. Exe + Gen resulted in serum Il-6 concentrations similar to that of controls. TNF-alpha concentrations did not differ by treatment. Overall, Exe had a positive impact on microbial richness, and Ruminococcus might be the driving bacteria for the GM structure differences. Exe + Gen may be an effective treatment for preventing HFD-induced inflammation.

Dysregulated bile acid receptor-mediated signaling and IL-17A induction are implicated in diet-associated hepatic health and cognitive function

BackgroundChronic consumption of high sugar and high fat diet associated with liver inflammation and cognitive decline. This paper tests a hypothesis that the development and resolution of diet-induced nonalcoholic fatty liver disease (NAFLD) has an impact on neuroplasticity and cognition.MethodsC57BL/6 wild-type mice were fed with either a healthy control diet (CD) or a fructose, palmitate, and cholesterol (FPC)-enriched diet since weaning. When mice were 3-months old, FPC diet-fed mice were randomly assigned to receive either FPC-enriched diet with or without 6% inulin supplementation. At 8 months of age, all three groups of mice were euthanized followed by analysis of inflammatory signaling in the liver and brain, gut microbiota, and cecal metabolites.ResultsOur data showed that FPC diet intake induced hepatic steatosis and inflammation in the liver and brain along with elevated RORγ and IL-17A signaling. Accompanied by microglia activation and reduced hippocampal long-term potentiation, FPC diet intake also reduced postsynaptic density-95 and brain derived neurotrophic factor, whereas inulin supplementation prevented diet-reduced neuroplasticity and the development of NAFLD. In the gut, FPC diet increased Coriobacteriaceae and Erysipelotrichaceae, which are implicated in cholesterol metabolism, and the genus Allobaculum, and inulin supplementation reduced them. Furthermore, FPC diet reduced FXR and TGR5 signaling, and inulin supplementation reversed these changes. Untargeted cecal metabolomics profiling uncovered 273 metabolites, and 104 had significant changes due to FPC diet intake or inulin supplementation. Among the top 10 most affected metabolites, FPC-fed mice had marked increase of zymosterol, a cholesterol biosynthesis metabolite, and reduced 2,8-dihydroxyquinoline, which has known benefits in reducing glucose intolerance; these changes were reversible by inulin supplementation. Additionally, the abundance of Barnesiella, Coprobacter, Clostridium XIVa, and Butyrivibrio were negatively correlated with FPC diet intake and the concentration of cecal zymosterol but positively associated with inulin supplementation, suggesting their benefits.ConclusionTaken together, the presented data suggest that diet alters the gut microbiota and their metabolites, including bile acids. This will subsequently affect IL-17A signaling, resulting in systemic impacts on both hepatic metabolism and cognitive function.

Neuroprotective effects of vildagliptin on drug induced Alzheimer's disease in rats with metabolic syndrome: Role of hippocampal klotho and AKT signaling pathways

Growing evidences suggest the presence of several similarities in the molecular mechanisms underlying the neurodegenerative diseases and metabolic abnormalities. Adults who develop Metabolic Syndrome (MS) are at a higher risk of developing Alzheimer's disease (AD). Pharmacological agents, like dipeptidyl peptidase-4 (DPP-4) inhibitors that increase the levels of glucagon like peptide 1 (GLP-1) and ameliorate symptoms of MS, have become an auspicious candidate as disease modifying agents in the treatment of AD. The present study investigates the beneficial effects of Vildagliptin, a DPP-4 inhibitor in counteracting cognitive decline in different models of dementia targeting the AKT, JAK/STAT signaling pathways and hippocampal Klotho expression, to judge the neuroprotective, anti-apoptotic and anti-inflammatory effects of the drug. Cognitive decline was induced by either administration of high fat high sugar (HFHS) diet for 45 days alone, or with oral administration of AlCl3 (100 mg/kg/day) for 60 days. Rats were orally administered Vildagliptin (10 mg/kg) for 60 days along with AlCl3 administration. Vildagliptin treatment improved spatial memory and activities in morris water maze (MWM) test and open field test respectively. Results revealed an increase of both hippocampal klotho and Bcl-2 expressions along with an increase in both AKT and ERK1/2 phosphorylation. In contrast, Vildagliptin treatment decreased hippocampal contents of inflammatory, apoptotic and oxidative stress biomarkers as TNF-α, caspase-3 and FOXO1 along with restoring metabolic abnormalities. A significant decrease in BAX expressions with JAK2/STAT3 inhibition was observed. These findings demonstrate that the neuroprotective role of vildagliptin is possibly via modulating Klotho protein together with AKT pathway.

Long-term intake of 9-PAHPA or 9-OAHPA modulates favorably the basal metabolism and exerts an insulin sensitizing effect in obesogenic diet-fed mice.

Fatty acid esters of hydroxy fatty acids (FAHFAs) are a large family of endogenous bioactive lipids. To date, most of the studied FAHFAs are branched regioisomers of Palmitic Acid Hydroxyl Stearic Acid (PAHSA) that were reported to possess anti-diabetic and anti-inflammatory activity in humans and rodents. Recently, we have demonstrated that 9-PAHPA or 9-OAHPA intake increased basal metabolism and enhanced insulin sensitivity in healthy control diet-fed mice but induced liver damage in some mice. The present work aims to explore whether a long-term intake of 9-PAHPA or 9-OAHPA may have similar effects in obesogenic diet-fed mice. C57Bl6 mice were fed with a control or high fat-high sugar (HFHS) diets for 12 weeks. The HFHS diet was supplemented or not with 9-PAHPA or 9-OAHPA. Whole-body metabolism was explored. Glucose and lipid metabolism as well as mitochondrial activity and oxidative stress status were analyzed. As expected, the intake of HFHS diet led to obesity and lower insulin sensitivity with minor effects on liver parameters. The long-term intake of 9-PAHPA or 9-OAHPA modulated favorably the basal metabolism and improved insulin sensitivity as measured by insulin tolerance test. On the contrary to what we have reported previously in healthy mice, no markedeffect for these FAHFAs was observed on liver metabolism of obese diabetic mice. This study indicates that both 9-PAHPA and 9-OAHPA may have interesting insulin-sensitizing effects in obese mice with lower insulin sensitivity.

Lepidium meyenii Supplemented Diet Modulates Neurobehavioral and Biochemical Parameters in Mice Fed High-Fat High-Sugar Diet.

Metabolic syndrome has been associated with an increased risk of cardiovascular disease, diabetes mellitus, and neurodegenerative disorders. Known side-effects of currently- available drugs necessitate the search for possibly better treatment options. This study examined the effects of dietary lepidium meyenii (MACA) supplementation on neurobehaviour, metabolic profile, levels of inflammatory markers, and oxidative stress parameters in a mouse model of metabolic syndrome. Mice were randomly assigned into 8 groups of ten animals each. Groups consist of standard diet (SD) control, high fat/high sugar (HFHS) control and three groups each of lepidium meyenii incorporated into either SD or HFHS diet at 0.1, 0.2 and 0.4%. Mice were fed for seven weeks, and body weight was measured weekly. Open-field behaviors and radial-arm/Y-maze spatial memory were scored at the end of the study. Twenty-four hours after the last behavioral test, fasting blood glucose levels were estimated. Animals were then euthanized, and blood was drawn for estimation of serum lipid profile. Whole brains were excised, weighed and homogenized to estimate the levels of lipid peroxidation, inflammatory markers, antioxidant status, and acetylcholinesterase activity. MACA-supplemented diet was associated with a decrease in body weight gain, an increase in food intake (at lower concentrations), suppression of grooming behavior, and decrease in acetylcholinesterase activity. MACA-supplement also reversed HFHS-induced memory impairment, anxiety, hyperglycaemia, lipid derangement, oxidative stress, and derangement of inflammatory markers. Dietary supplementation with MACA shows beneficial effects in mitigating the effects of metabolic syndrome on the brain in mice.

Ghrelin-induced multi-organ damage in mice fed obesogenic diet.

Ghrelin has a key role in modulating energy metabolism and weight gain. The present study aimed at studying the potential role of ghrelin in the development and/or exacerbation of organ damage in a mouse model of diet-induced obesity. Adult mice were fed one of two diets for 20weeks: standard high carbohydrate (HC) or high-fat high-sugar (HFHS). Starting week 17, the animals were given regular intraperitoneal ghrelin (160µg/kg) or saline injections Abdominal fat, serum creatinine, and glucose levels, as well as kidney, liver and heart weight and pathology were assessed. Ghrelin-injected mice showed significant organ damage, which was more exacerbated in HFHS-fed animals. While the HFHS diet was associated with significant liver damage, ghrelin administration did not reverse it. Interestingly, ghrelin administration induced moderate kidney damage and significantly affected the heart by increasing perivascular and myocardium fibrosis, steatosis as well as inflammation. Moreover, serum creatinine levels were higher in the animal group injected with ghrelin. Ghrelin administration was associated with increased functional and structural organ damage, regardless of diet. The present study provides novel evidence of multi-organ physiologic alterations secondary to ghrelin administration.

Intact vagal gut-brain signalling prevents hyperphagia and excessive weight gain in response to high-fat high-sugar diet.

AimThe tools that have been used to assess the function of the vagus nerve lack specificity. This could explain discrepancies about the role of vagal gut‐brain signalling in long‐term control of energy balance. Here we use a validated approach to selectively ablate sensory vagal neurones that innervate the gut to determine the role of vagal gut‐brain signalling in the control of food intake, energy expenditure and glucose homoeostasis in response to different diets.MethodsRat nodose ganglia were injected bilaterally with either the neurotoxin saporin conjugated to the gastrointestinal hormone cholecystokinin (CCK), or unconjugated saporin as a control. Food intake, body weight, glucose tolerance and energy expenditure were measured in both groups in response to chow or high‐fat high‐sugar (HFHS) diet. Willingness to work for fat or sugar was assessed by progressive ratio for orally administered solutions, while post‐ingestive feedback was tested by measuring food intake after an isocaloric lipid or sucrose pre‐load.ResultsVagal deafferentation of the gut increases meal number in lean chow‐fed rats. Switching to a HFHS diet exacerbates overeating and body weight gain. The breakpoint for sugar or fat solution did not differ between groups, suggesting that increased palatability may not drive HFHS‐induced hyperphagia. Instead, decreased satiation in response to intra‐gastric infusion of fat, but not sugar, promotes hyperphagia in CCK‐Saporin‐treated rats fed with HFHS diet.ConclusionsWe conclude that intact sensory vagal neurones prevent hyperphagia and exacerbation of weight gain in response to a HFHS diet by promoting lipid‐mediated satiation.

EFFECTS OF DIET ALTERATIONS, WITH OR WITHOUT FECAL MICROBIAL TRANSPLANTS, ON BONE INTEGRITY

High fat feeding exerts a negative impact on bone quality and strength. Gut microbiota have been strongly linked to bone outcomes in several models, though a clear mechanism linking alterations in gut microbiota, diet composition, and bone has not yet been elucidated. PURPOSE: To determine if alterations in diet, with or without fecal microbial transplants (FMT) can rescue bone integrity in diet-induced obesity. METHODS: 6-wk old male C57Bl/6 mice (n=10/group) were randomized to a low-fat (LF) or high fat, high sugar (HFS) diet ad libitum for 13 wks. HFS mice were randomized to one of three groups for 4 wks: LF diet with FMT from the LF mice (HFS/LF+), LF diet with sham FMT using PBS (HFS/LF), or HFS diet with FMT from the LF mice (HFS/HFS+) to simulate the impact of combined diet alteration and addition of “healthy” microbes, diet alterations only, and addition of “healthy” microbes only, respectively. Animals had free access to a running wheel until terminated at 23 wks of age. Statistical analyses were performed using a two-way ANOVA and Tukey’s post-hoc test. RESULTS: HFS/HFS+ mice showed greater absolute femoral neck (FN) strength versus HFS/LF and HFS/LF+ mice (p<0.003), while LF/LF mice had the greatest relative FN strength versus all groups (p<0.016). Whole femur bone mineral density (BMD, g/cm2) was greater in HFS/HFS+ versus all groups (p<0.023); LF/LF mice had the highest BMD when normalized to body weight (p<0.029). Immunostaining for tumor necrosis factor alpha, sclerostin, insulin-like growth factor 1, and interferon gamma in cortical and cancellous bone revealed no differences between groups. Despite no difference in marrow adipocyte number (#/mm2) between groups, HFS/HFS+ mice had greater marrow adipocyte size (μm2) versus LF/LF mice; diet alteration, with and without transplanted “healthy” microbes, was able to partially attenuate increased marrow adipocyte size. CONCLUSIONS: A change in diet from HFS to LF led to reduced absolute FN strength compared to HFS/HFS+ and LF/LF mice, with no apparent effect from transplanted “healthy” gut microbes. LF/LF mice had greater relative femoral neck strength. Altering diet, with and without FMT’s, was sufficient to partially rescue the detrimental impacts of high fat, high sugar feeding on bone integrity.

Expression Of Tyrosine Hydroxylase In The Nucleus Accumbens Are Not Altered By Diet Or Fecal Transplantation In Male C57bl/6j Mice

Multiple studies have indicated that regulation of physical activity may be largely controlled by central neural factors, such as dopamine (DA) signaling in the nucleus accumbens (NAc). It was hypothesized that metabolites produced from the gut microbiome influence DA signaling by altering the rate-limiting enzyme tyrosine hydroxylase (TH). PURPOSE: To determine whether microbial transplants from high active C57Bl/6J male mice eating a chow diet (CH) altered TH expression in the NAc of C57Bl/J male mice made low active through intake of a high fat high sugar (HF) diet. METHODS: Mice were randomly assigned to one of four groups: a chow diet (CH/CH), a high fat to chow diet plus a microbial transplant (HF/CH+), a high fat diet to chow diet (HF/CH), or a high fat diet plus a microbial transplant (HF/HF+). Changes to group base diets and microbial transplants began at week 14. Transplants were completed once a week using sample donated from the CH/CH group via oral gavage. Mice were sacrificed at the end of 17 weeks, the NAc was dissected on ice, and flash frozen in liquid nitrogen. Immunoblotting was performed using NAc lysate probed with TH antibody. Bands were normalized using β-actin and an analysis of variance was conducted. RESULTS: There were no significant differences between group means for CH/CH (1.023 ± 0.59), HF/CH+ (0.8810± 0.21), HF/CH (0.882± 0.27), or HF/HF+ (1.069± 0.32) as determined by a one-way ANOVA, (p = 0.51). CONCLUSION: TH expression in the NAc was not altered by diet or microbial transplantation from the active CH/CH group. Funding for this study was provided by the Omar Smith Endowment at Texas A&M University.

Effect Of Aerobic Exercise On Cardiac Diastolic Dysfunction: Role Of Heat Shock Protein 27

PURPOSE: To evaluate the role of heat shock protein 27 (HSP27) in the effect of aerobic exercise on cardiac diastolic dysfunction in type 2 diabetic rats. METHODS: Forty male Sprague-Dawley rats were randomly divided into four groups: control (C), aerobic exercise (A), diabetes (D), and diabetes plus aerobic exercise (DA). Type 2 diabetes was induced by feeding with a high-fat high-sugar diet for 7 weeks followed by a single intraperitoneal injection of streptozotocin (30mg/kg) in the rats. Aerobic exercise was performed on a rodent treadmill at 21m/min for 60 min, 5 days per week for 8 weeks. Metabolic factors, such as fasting blood glucose (FBG), triglycerides (TG), cholesterol (CHOL) and lipoproteins, were determined by a standard procedure. Cardiac structure and function, such as left ventricular diastolic upper diameter (LVIDd), left ventricular diastolic terminal volume (EDV) and ejection fraction (EF), were measured using echocardiography. Myocardial HSP27 protein expression and phosphorylation were determined using western blot, and myocardial HSP27-titin colocalization were determined using double immunofluorescence. Two-way ANOVAs with post-hoc tests were used to assess differences between groups. RESULTS: No pathological changes were observed in the myocardial structure in all four groups. Compared to the C group, the D group had significantly higher levels of FBG, TG, and CHOL (all p<0.01), and significantly lower LVIDd and EDV (both p<0.01). Compared to the D group, the DA group had significantly lower levels of FBG, TG, and CHOL (all p<0.01) and significantly higher LVIDd and EDV (both p<0.05). Interestingly, compared to the C group, the D group showed significantly lower myocardial HSP27 phosphorylation and HSP27-titin colocalization (both p<0.05), while the DA group showed significantly higher myocardial HSP27 protein expression and phosphorylation, and HSP27-titin colocalization than the D group (all p<0.01). CONCLUSION: Decreases in myocardial HSP27 phosphorylation and HSP27-titin colocalization are likely involved in early diastolic dysfunction in diabetic rats. Increased HSP27 phosphorylation and titin colocalization may be an important mechanism by which aerobic exercise restores diastolic function in diabetic rats.

Nullification of Hepatic Cannabinoid 1 Receptor (CB1R) Ameliorates Fatty Liver and Improves Glucose Homeostasis via Regulation of PI3K-mTORC1 axis

Abstract Objectives Cannabinoid 1 receptors (CB1Rs) are G protein-coupled receptors that are present in peripheral organs such as liver, pancreas, adipose tissue and skeletal muscle where they are involved in fine-tuning many metabolic functions. It was previously reported that liver-specific genetic deletion of CB1R (hCNR1−/−) mice fed high fat diets (HFD) had similar body weight to HFD-fed hCNR1+/+ mice, but they retained insulin sensitivity comparable to normal chow-fed (NCD) hCNR1+/+ mice. Therefore, this study was undertaken to uncover how hCB1Rs impact the insulin signaling pathway. Methods Male hCNR1−/− and hCNR1+/+ mice were fed with a high fat high sugar diet (HFSD: N = 8–12) for 15 weeks. Results In contrast with the previous study of HFD only, HFSD-fed hCNR1−/− mice had less body weight gain than HFSD-fed hCNR1+/+ mice (25.1 ± 2.7 vs 19.8 ± 1.7 g), less fat accumulation in the liver, 1.3-fold increase in glucose disposal and 1.7-fold increase in insulin sensitivity compared to HFSD-fed hCNR1+/+ mice. Further study demonstrated that the lack of hepatic CB1R resulted in upregulated phosphorylation in liver of protein kinase B (AKT), causing activation of downstream target molecules, such as proline-rich AKT substrate 40 (PRAS40) in the mammalian target of rapamycin complex 1 (mTORC1). Similarly, primary hepatocytes isolated from hCNR1−/− mice had increased amounts of phosphorylated AKT and PRAS40 in comparison to hepatocytes from hCNR1+/+ mice. The co-immunoprecipitation studies clearly revealed hepatic CB1R modulates insulin signaling by the association or dissociation of the components of mTORC1. In support of these data, the deficiency of hepatic CB1R leads to upregulation of phosphoinositide 3-kinase (PI3K) class IB, p110γ, resulting in increased association between p110γ and G-protein beta. Conclusions These findings indicate that genetic deletion of hepatic CB1R contributes to improved glucose homeostasis. Therefore, modulation of CB1R activity in liver may be a useful therapeutic in obese and diabetic individuals. Funding Sources Intramural Research Program of the National Institute on Aging.

The experimental application of steam explosion-pre-treated flaxseed meal with hypoglycaemic and lipid-lowering functions in rats on a high-fat-sugar diet

With the increase in populations with suboptimal health and chronic diseases, consumer demands for health foods, especially food products with health functions. The flaxseed was a complex mixture mainly consisting of flaxseed oil, flaxseed protein, dietary fibre (including cellulose and flaxseed gum), carbohydrates and secoisolariciresinol diglucoside (SDG). To investigate the effects of expanded flaxseed meal (EFM) intervention on the hypoglycaemic and lipid-lowering functions of rats on a high-fat, high-sugar (HFHS) diet, EFM was fed to six groups (z = 8) of Sprague–Dawley (SD) rats: normal control, HFHS model, positive treatment control, and low-, medium-, and high-dose EFM (5%, 10% and 20% mass fraction EFM, respectively) groups. The results (8 weeks later) were as follows: (1) medium- and high-dose EFM achieved similar inhibitive effects on blood glucose levels compared with positive treatment (5.94±0.32 mmol/L); (2) all EFM doses significantly reduced total cholesterol, triglyceride and low-density lipoprotein cholesterol levels in the blood plasma of SD rats and (3) EFM significantly increased glutathione peroxidase, superoxide dismutase and catalase activities, increased glutathione levels, and reduced the malondialdehyde content in the plasma of SD rats (P &lt; 0.05). Therefore, EFM mitigated the adverse effects produced by HFHS diet on physiological functions, effectively regulated blood glucose and lipid metabolism, enhanced plasma antioxidant capacity and alleviated oxidative stress in SD rats with a certain dose–effect relationship among the EFM dose groups. Hepatic histological morphologies indicated that EFM also improved the size and density of lipid droplets in the liver of the rats and inhibited the onset of fatty liver. These results suggest that EFM is a potentially useful raw material in the development of blood glucose and lipid-lowering food products.

Baricitinib counteracts metaflammation, thus protecting against diet-induced metabolic abnormalities in mice

ObjectiveRecent evidence suggests the substantial pathogenic role of the Janus kinase (JAK)/signal transducer and activator of transcription (STAT) pathway in the development of low-grade chronic inflammatory response, known as “metaflammation,” which contributes to obesity and type 2 diabetes. In this study, we investigated the effects of the JAK1/2 inhibitor baricitinib, recently approved for the treatment of rheumatoid arthritis, in a murine high-fat-high sugar diet model. MethodsMale C57BL/6 mice were fed with a control normal diet (ND) or a high-fat-high sugar diet (HD) for 22 weeks. A sub-group of HD fed mice was treated with baricitinib (10 mg/kg die, p.o.) for the last 16 weeks (HD + Bar). ResultsHD feeding resulted in obesity, insulin-resistance, hypercholesterolemia and alterations in gut microbial composition. The metabolic abnormalities were dramatically reduced by chronic baricitinib administration. Treatment of HD mice with baricitinib did not change the diet-induced alterations in the gut, but restored insulin signaling in the liver and skeletal muscle, resulting in improvements of diet-induced myosteatosis, mesangial expansion and associated proteinuria. The skeletal muscle and renal protection were due to inhibition of the local JAK2-STAT2 pathway by baricitinib. We also demonstrated that restored tissue levels of JAK2-STAT2 activity were associated with a significant reduction in cytokine levels in the blood. ConclusionsIn summary, our data suggest that the JAK2-STAT2 pathway may represent a novel candidate for the treatment of diet-related metabolic derangements, with the potential for EMA- and FDA-approved JAK inhibitors to be repurposed for the treatment of type 2 diabetes and/or its complications.

SAT-659 Micro/Nanoparticles from Antidiabetic Chinese Herbal Medicine Soup Increase Bioavailability of Phytochemicals and Regulate Glucose and Lipid Metabolism

Herbal soups have always been a popular option for treating oxidative stress-related chronic diseases including diabetes. Various components of these soups have been studied in the hope to identify the active principles, mainly focusing on the individual phytochemicals. As we have revealed previously, the micro/nanoparticles (MNPs) formed incidentally during the boiling water extraction of herbal soups may be bioactive and functional. This study aims to elucidate the underlying mechanism of the biological functions of these MNPs. A Chinese herbal medicine soup prepared by Radix Puerariae lobatae, ginger and other three herbs, namely Ge-Gen-Qin-Lian-Tang, was employed here, as it was proven to be effective in treating type 2 diabetes clinically.The soup was separated with high-speed centrifuge (15600×g) to obtain the supernatant (solutes and nanoparticles) and sediments (MNPs), and determined for the content of three bioactive phytochemicals, e.g. puerarin, berberine and baicalin. Their hypoglycemic, hypolipidemic and antioxidant effects were determined on streptozotocin (STZ)-induced Type 2 diabetic Wistar male rats fed on high fat-high sugar diet. The animals were divided into six groups (normal control, diabetic model, whole soup, supernatant, MNPs and metformin, 8 rats each), recording weight, diet, excretion, mental status, etc. The fasting blood glucose and oral glucose tolerance test were conducted regularly. Eight weeks after the administration, the rats were sacrificed after anesthesia. Abdominal aorta blood and tissue samples of pancreas, heart, skeletal muscle, liver, kidney, spleen were collected. The glycated hemoglobin, glucose, lipid, insulin, glucagon, AMPK, SOD, puerarin, berberine, baicalin in blood plasma, insulin in pancreas, SOD in tissues, AMPK in skeletal muscle were measured. Liver tissue sections were observed with HE staining. Statistical analysis (t-test) were performed.The MNPs reduced blood glucose, ameliorated glucose tolerance, elevated insulin secretion and significantly improve glucose and lipid metabolism (P <0.05), showing stronger effects than the supernatant components. Notably, MNPs elevated the AMPK level in skeletal muscle, even more potently than the whole soup. The therapeutic effects of MNPs on the liver damage were even stronger than metformin. Meanwhile, MNPs promoted absorption of puerarin, berberine and baicalin and increase their concentration in blood (P <0.05). Therefore, the MNPs from the herbal soup exhibited more potent effects than the soluble components on ameliorating glucose and lipid metabolism and pancreatic functions of diabetic rats. The actions of these MNPs provide a new perspective for understanding the antidiabetic effects of herbal soups and serve as a vehicle for the multiple phytochemicals to synergistically possess therapeutic effects.

Sex differences in response to a high fat, high sucrose diet in both the gut microbiome and hypothalamic astrocytes and microglia

ABSTRACT Objectives: Obesity is a major epidemic in our population and has emerged as a primary health concern. Consumption of a high fat, high sugar (HFHS) diet can specifically lead to gut dysbiosis, increased inflammation, and neuroinflammation. Interestingly, sex differences in the response to a HFHS diet are emerging. In this study, we investigated the effects of a HFHS diet compared to a low fat, low sugar (LFLS) diet in 8 week old male and female C57Bl/6 mice. Methods: The diet was administered for 14 weeks; body weights and food consumption were evaluated weekly. Results: Male and female mice fed the HFHS diet gained significantly more weight than LFLS-fed mice. However, in agreement with previous studies, males gained significantly more weight on the HFHS diet compared to females fed the same diet. Importantly, we determined significant sex and diet-induced differences to gut microbiome composition using next generation Illumina sequencing. We also observed significantly less astrocyte densitometry and no significant change to microglial morphology in the hypothalamus of Female HFHS compared to Female LFLS. On the other hand, Male HFHS revealed no change to hypothalamic astrogliosis, but increased microgliosis compared to Male LFLS. Discussion: In this study, we determined sex and diet-induced differences in both the gut and the brain, however, future studies will need to be performed in order to test the direct role of the gut microbiome to weight gain and neuroinflammation in male and female mice.

Influence of exercise and/or genistein on gut microbiota after 12 weeks of Western diet in C57BL/6 adult mice

Diet and exercise have been studied independently as potential treatments to revert the negative effects of Western diets on host health through the modification of gut microbiota. The potential synergic effect of diet and exercise has not been well studied. This study aimed to analyze the impact of exercise (Exe), genistein (Gen), and their combined effect on gut microbiota after 12 wks of a high‐fat, high‐sugar diet (HFD).MethodsEighty‐three C57BL/6 mice (male n=41, female n=42) aged 4–5 wks were randomized to one of the following groups for a 12‐wk period: control (CON), HFD, HFD + Exe, HFD + Gen, or HFD + Exe + Gen. The HFD diet contained 20% carbohydrate, 20% protein, and 60% fat with sugar‐supplemented drinking water (42 g sugar/L). Gen containing diets had 600 mg Gen/kg of diet. Exe groups underwent moderate‐intensity aerobic exercise 30 min/d for 5 d/wk (wk 1 at 10 m/min for 15 min, wk 2 at 10 m/min increased to 30 min, wks 3–12 12 m/min for 30 min). Microbial DNA was extracted from fecal pellets collected at 12 wks and sequenced on the Illumina miSeq platform using primers for the V4 region of the 16S rRNA gene. The QIIME 2019.7 pipeline was used to align genomic sequences (DADA2) and assign taxonomy (Greengenes 13_8 database). a‐ (Shannon, Observed Operational Taxonomical Units (Ob OTUs), Faith PD, Pielou’s evenness) and b‐diversity (Jaccard, Bray‐Curtis, weighted and unweighted UniFrac) metrics were compared (Kruskal‐Wallis (KW) sum‐rank test and pairwise permutational multivariate analysis of variance, respectively) by treatment group. All‐against‐all LEfSe analysis was used to identify differential microbial abundance group differences (KW sum‐rank test).ResultsMicrobial abundance (Shannon, Pielou’s Evenness) did not differ by study group but the CON group had a higher species richness (Ob OTUs, Faith PD; p&lt;0.001) after 12 wks when compared to all other groups. HFD + Exe, and HFD + Gen + Exe groups also had a higher microbial richness (Ob OTUs, Faith PD; p&lt;0.01) when compared to HFD + Gen suggesting that exercise may increase the number of unique microbes present in the community. All b‐diversity metrics suggested that the microbial community structure differed significantly (p&lt;0.001), such that Exe, Gen, and Gen + Exe treatments each had unique effects on microbial diversity. LEfSe analysis (LDA score &gt; 2; p&lt;0.05) identified that the genus Ruminococcus was more abundant in both CON and HFD + Exe + Gen groups, when compared to all other groups.ConclusionThese data suggest that exercise may have a greater impact on species richness than genistein but that each treatment elicits unique effects on the overall microbial community structure. Abundance of Ruminococcus, a lactic acid producer, common among CON mice was rescued only in HFD + Exe + Gen mice. The lack of an effect with Exe or Gen alone is interesting given that Ruminococcus has independently been shown to convert Gen to active secondary metabolites and has also been positively associated with Exe. This may suggest that a synergistic approach is required to overcome the negative gut microbial effects of a HFD. Further study is needed to understand how gut microbiome changes relate to metabolic health outcomes.Support or Funding InformationMidwestern Arizona Alzheimer’s Consortium (AZ0049)Alpha diversity boxplot for Observed OTUs.(*) denotate significant group differences (p&lt;0.05) after Benjamini‐Hochberg correction for multiple comparisons. Treatment groups are as follows: (*) Control differed to HFD and HFD + Gen; (**) HFD vs HFD + Exe; (***) HFD vs HFD + Exe + Gen. High Fat Diet (HFD); (****) HFD+ Exe vs HFD + Gen; (*****) HFD + Exe + Gen vs HFD+ gen. Exercise (Exe); Genistein (gen).Figure 1Effect size of differentially abundant genera of Rominococcus all‐against‐all Linear discriminate analysis of effect size (LEfSe) between a) Control, High‐Fat Diet (HFD), HFD + Exercise (Exe), HFD +Genistein (Gen), and HFD + Exe + Gen groups b) between High‐Fat Diet (HFD), HFD + Exercise (Exe), HFD +Gcnistein (Gen), and HFD + Exe + Gen groups The phylogenetic features were tested by LEfSe to reveal the genera which significantly differed between the different groups.Figure 2

Role Of SDF:CXCR4 In Diet‐Induced Cardiac Injury

IntroductionDiabetic cardiomyopathy (DCM) is a major cardiovascular complication in patients with diabetes and is defined as ventricular dysfunction (in diabetes) independent of coronary artery disease. In this study, we define a novel role for the SDF‐1: CXCR4 axis in diabetes‐associated myocardial dysfunction.MethodsWild‐type mice were randomly assigned to a high fat high sugar diet (HFHS) or control diet (LF) for 14 months. Serial echocardiography was used to assess cardiac function. The hSDF‐1 plasmid was injected into the LV wall of HFHS mice 7 months and 14 months after HFHS diet. The mRNA levels in the hearts were quantified by qPCR.ResultsHFHS‐fed mice (vs. control diet) showed significantly increased deceleration time (diastolic dysfunction) 7 months after HFHS diet and decreased Ejection Fraction (EF) (systolic dysfunction) 14 months after the HFHS diet. We observed a significant increase in cardiac myocytes surface area and a decrease in vascular density in the hearts of HFHS mice. HFHS‐fed mice showed decreased P‐selectin, E‐selectin expression and increased CXCR4, Resistin, and DDR2 expression in the heart. There is no significant improvement in diastolic function after the SDF‐1 injection at 7 month. However, direct myocardial injection of hSDF‐1 plasmid led to a significant improvement in EF compared to the control group. To determine the role of CM CXCR4 in hyperglycemia associated cardiac injury, we quantified cardiac function and survival rate in HFHS‐fed CM‐CXCR4 null mice. We observed a significantly increased mortality rate before 14 months on HFHS fed CM‐CXCR4 null mice compare to HFHS‐fed control mice.ConclusionHFHS diet induces diastolic dysfunction in the short term then systolic dysfunction after a long term. SDF‐1 treatment ameliorates systolic dysfunction but not diastolic dysfunction. Our data suggest that cardiac myocyte expression of CXCR4 has an important role in diabetes‐associated cardiac injury.

A43 β-DEFENSINS AS MARKERS OF INTESTINAL DYSBIOSIS: THE NATURE OF CHANGES IN β-DEFENSINS IS DEPENDENT ON THE PROCESS UNDERLYING THE INDUCTION OF DYSBIOSIS

Abstract Background Dysbiosis may be defined as a change in the microbial composition or function that results in altered host function. Defensins are antimicrobial peptides, are part of innate immunity, and are important in host defense and maintaining homeostasis. Dysbiosis is a putative mechanism underlying the expression of many functional GI disorders like Irritable Bowel Syndrome (IBS) for which no biomarkers exist. Previous studies have revealed increased β-defensin (β-Def) levels in IBS patients, most likely due to changes in the microbiota. Aims We examined the hypotheses that: 1) Changes in β-Def are dependent on the manner in which dysbiosis is induced, and that 2) the direction of the change in β-Def depends on how dysbiosis was induced. Methods We used 4 models of experimentally induced dysbiosis to determine changes in fecal β-Def and to characterize the microbiota composition before and during the induction of dysbiosis. We used: 1) an antimicrobial cocktail (AC) in water; 2) a high-fat/ high-sugar diet (HFHSD); 3) a high salt diet (HSD) that we previously showed to induce a pro-inflammatory microbiota; and 4) mild restraint stress (MRS). All studies were performed in C57/BL6 mice except studies using MRS that were performed in NIH Swiss mice. In the AC or dietary studies, we employed a one-week intervention preceded by one-week baseline and recovery periods. In MRS studies, mice comparisons were made between a control and a stressed group. Stool samples were collected every 24 hours and were assayed for fecal β-Def levels analysis by an ELISA and microbial composition by 16S gene profiling. Results Exposure to AC or dietary change, but not MRS, resulted in significant decreases in fecal β-Def. Additionally, bacterial composition and diversity profiles were different in all mice except MRS mice (control vs. MRS males: p=0.414; control vs. MRS females: p=0.96). In contrast, mice exposed to the HSD revealed a significant increase in β-Def during treatment compared to baseline in both males (p=0.025) and females (p= 0.0019). The AC mice showed the largest changes and significant correlations between changes in β-Def levels and bacterial diversity (males: p=0.013, r=0.6; females: p=0.007, r=0.6) and richness (males: p=0.0008, r=0.70; females: p=0.003, r=0.62). However, no significant correlations were found between specific bacteria and β-Def levels in the HFHSD group. Conclusions We conclude that directional changes in fecal β-Def levels are dependent on the manner in which dysbiosis is induced. The use of β-Def as a biomarker requires comparisons with baseline levels obtained during remission in order to identify dysbiosis presence in microbiota-associated chronic GI conditions like IBS. Such an approach will identify patient subgroups that may benefit from microbiota-directed therapies. Funding Agencies CIHR