High-fat Diet Animals Research Articles

Oral Administration of CardioAid and Lunasin Alleviates Liver Damage in a High-Fat Diet Nonalcoholic Steatohepatitis Model

Background: Several of the drugs in development for treatment of nonalcoholic steatohepatitis (NASH) target liver fibrosis or have side effects that prohibit their long-term use in patients with mild to moderate disease. Lunasin is a soy-derived peptide with anti-inflammatory properties. ADM's CardioAid™ is a plant sterol extract that exerts cholesterol- and triacylglycerol-lowering effects. Aim: To determine the immunomodulatory effects of CardioAid and lunasin in a high-fat diet (HFD) animal model of NASH. Methods: C57BL/6 mice on an HFD were orally administered CardioAid or lunasin for 25 weeks. The effects on the immune system, liver function, insulin resistance and lipid profile were studied. Results: Treatment with CardioAid and lunasin was associated with a significant decrease in the CD4/CD8 ratio and an increase in CD4+CD25+ lymphocytes. A decrease in interleukin 1-alpha serum levels and an increase in transforming growth factor beta serum levels were noted. These were associated with alleviation of liver damage as indicated by a significant decrease in liver enzymes and improvement in the histological nonalcoholic fatty liver disease activity score (NAS). Decreases in both serum triglyceride and serum glucose levels were observed in treated mice. A decrease in total body fat measured by EchoMRI was also observed in treated mice. Conclusions: CardioAid and lunasin exerted hepatoprotective and glucose-protective effects in an HFD NASH model. These data and the high-safety profiles of CardioAid and Lunasin support their use in patients in the early stages of NASH to prevent deterioration due to the disease.

Omega-3 fatty acids revert high-fat diet-induced neuroinflammation but not recognition memory impairment in rats

Neuroinflammation is a consequence of overeating and may predispose to the development of cognitive decline and neurological disorders. This study aimed to evaluate the impact of omega-3 supplementation on memory and neuroinflammatory markers in rats fed a high-fat diet. Male Wistar rats were divided into four groups: standard diet (SD); standard diet + omega-3 (SD+O); high fat diet (HFD); and high fat diet + omega-3 (HFD+O). Diet administration was performed for 20weeks and omega-3 supplementation started at the 16th week. HFD significantly increased body weight, while omega-3 supplementation did not modify the total weight gain. However, animals from the HFD+O group showed a lower level of visceral fat along with an improvement in insulin sensitivity following HFD. Thus, our results demonstrate a beneficial metabolic role of omega-3 following HFD. On the other hand, HFD animals presented an impairment in object recognition memory, which was not recovered by omega-3. In addition, there was an increase in GFAP-positive cells in the cerebral cortex of the HFD group, showing that omega-3 supplementation can be effective to decrease astrogliosis. However, no differences in GFAP number of cells were found in the hippocampus. We also demonstrated a significant increase in gene expression of pro-inflammatory cytokines IL-6 and TNF-α in cerebral cortex of the HFD group, reinforcing the anti-inflammatory role of this family of fatty acids. In summary, omega-3 supplementation was not sufficient to reverse the memory deficit caused by HFD, although it played an important role in reducing the neuroinflammatory profile. Therefore, omega-3 fatty acids may play an important role in the central nervous system, preventing the progression of neuroinflammation in obesity.

Capsanthin Inhibits both Adipogenesis in 3T3-L1 Preadipocytes and Weight Gain in High-Fat Diet-Induced Obese Mice.

Adipogenesis in murine preadipocyte 3T3L-1 has been used as a model system to study anti-obese bioactive molecules. During adipogenesis in 3T3-L1 preadipocytes, we found that capsanthin inhibited adipogenesis (IC50; 2.5 μM) and also showed lipolytic activity in differentiated adipocytes from the preadipocytes (ED50; 872 nM). We identified that the pharmacological activity of capsanthin on adipogenesis in 3T3-L1 was mainly due to its adrenoceptor-β2-agonistic activity. In high-fat diet animal model study, capsanthin significantly enhanced spontaneous locomotive activities together with progressive weight-loss. The capsanthin-induced activation of kinetic behavior in mice was associated with the excessive production of ATP initiated by both the enhanced lipolytic activity together with accelerated oxidation of fatty acids due to the adrenoceptor β2-agonistic activity of capsanthin. Capsanthin also dose-dependently increased adiponectin and p-AMPK activity in high fat diet animals, suggesting that capsanthin has both anti-obesity and insulin sensitizing activities.

Effects Of Physical Exercise On The Modulation Of Aquaglyceroporin 7 From Visceral Adipose Tissue

A role for aquaglyceroporin 7 (AQP7) in the control of lipid accumulation in white adipose tissue (WAT) has been hypothesized; however limited information is available regarding the impact of physical exercise. PURPOSE: We aimed to analyze the role of voluntary physical activity (VPA) and endurance training (ET) on WAT AQP7 expression of HFD-fed rats. METHODS: Male Sprague-Dawleys rats were assigned into sedentary (S), VPA and ET groups fed an isoenergetic Lieber-DeCarli-liquid diet: a standard diet (35% fat-derived Kcal) or a high fat-diet (HFD), (70% fat-derived Kcal), as follows: SS, SVPA, SET, HS, HVPA and HET, during 17 wks. VPA groups had free access to running wheel throughout the entire protocol. After 9 weeks of HFD, SET and HET animals were submitted to 8-wks of ET on treadmill while maintained dietary treatments. Plasma non-esterified fatty acid (NEFA), glycerol and insulin levels were determined and epididymal white adipose tissue (eWAT) was used to determine gene and protein expression of AQP7 and fatty acid translocase (FAT/CD36). Diet and exercise effects were performed using 2-way ANOVA. RESULTS: The relative caloric intake was constant between groups. HFD increased visceral adiposity index (9.0±0.2 vs. 11.8±0.4; p<0.0001) and adipocyte area mean (3716.1±301.4 vs. 5348.6±471.5; p<0.001), ET reduced these obesity-related anatomical features (p<0.001). Both AQP7 and FAT/CD36 protein and gene expression remained unchanged after HFD regimen. VPA decreased plasma glycerol levels (12.6±0.8 vs. 12.5±0.6; p<0.001) and eWAT AQP7 gene expression (p<0.001) in S diet-fed animals and had no impact neither on plasma NEFA levels nor FAT/CD36 protein content. Eight-wks of ET decreased NEFA (SS vs. SET, 16.4±0.5 vs. 12.0±0.4; HS vs. HET, 14.7±0.7 vs. 10.8±0.7, p<0.001) and glycerol (SS vs. SET, 17.4±0.7 vs. 12.5±0.6; HS vs. HET, 15.6±0.5 vs. 10.3±0.9; p<0.001) plasma levels. ET decreased gene and protein expression of AQP7 (p<0.001) in eWAT in both diet types and increased FAT/CD36 gene (p<0.001) in SET group and its protein expression (p<0.001) in both diet types. CONCLUSION: Our data suggest that 8-wks of ET decreased AQP7 expression, which might play an important role preventing lipid overaccumulation in visceral adipose tissue of obese rats. Supported by: UID/DTP/00617/2013; POCI-01-0145-FEDER-016690

Gene therapy for follistatin decreases systemic inflammation and pain sensitivity following knee injury in high-fat diet induced obese mice

Purpose: Both obesity and joint trauma are risk factors for osteoarthritis (OA) and OA-related pain. In addition, muscle may have a role in knee OA development as changes in muscle strength could affect kinematics of joint loading. Furthermore, because of its key function in glucose storage and utilization, muscle strength and physiology are particularly vulnerable to obesity-associated inflammation. However, the mechanical, inflammatory, and metabolic links among obesity, muscle, and OA remain unclear. Follistatin (FST), an activin-binding protein, has been used as an effective treatment for muscle degenerative diseases by increasing muscle growth due to its ability to inhibit myostatin, a negative regulator of muscle mass. Moreover, FST has been reported to reduce the infiltration of inflammatory cells in the synovial membrane of the knee joint. Therefore, we hypothesized that overexpression of FST using a gene therapy approach will mitigate obesity-associated inflammation and pain sensitization in a high-fat diet (HFD) induced obesity model of mouse OA. Methods: Six-week old male C57BL/6 mice were fed either a chow control-diet or HFD (60% fat by kcal, n = 16/dietary group). At 7-weeks of age, either an AAV9-mediated FST 314 gene vector or GFP-labeled control vector (n = 10 for each dietary groups) was delivered via tail vein at a final dose of 6 × 1011 vg/ mouse. At 16-weeks of age, mice in each group (n = 10) underwent surgery to destabilize the medial meniscus (DMM) on the left knee. The remaining mice served as non-surgical controls. At time of surgery, Dual Energy X-ray Absorptiometry was conducted to measure body composition and bone mineral density (BMD). Six-weeks post-surgery (22-weeks of age), blood was collected and analyzed using a 31-plex Luminex® assay. Peripheral thermal sensitivity was determined using a hot/cold plate, while mechanical pain algesia at the knee was evaluated by a pressure-pain test. A two-way ANOVA between groups or a paired t-test within each group was performed to determine statistical significance, as appropriate (α = 0.05). Results: Prior to surgery, HFD resulted in increased body weight for both GFP- and FST-treated animals compared to chow (p < 0.05). However, overexpression of FST significantly decreased percentage of body fat in both chow and HFD treated mice as compared to their own corresponding GFP-controls. There was a main effect of both diet (p = 0.028) and FST-treatment (p = 0.007) on bone mineral density, such that HFD decreased BMD (HFD-GFP: 0.46 ± 0.23g/cm2 vs. chow-GFP: 0.51 ± 0.15g/cm2) while FST increased BMD (HFD-FST: 0.52g/cm2 ± 0.17, chow-FST: 0.56g/cm2 ± 0.17). Six weeks post-surgery, serum inflammatory profiles for HFD-GFP animals demonstrated increased systemic inflammation compared to chow diet, which was attenuated in the HFD-FST animals across fourteen markers (Table 1). HFD increased hot-plate paw-withdrawal latency time in GFP-treated animals compared to all other groups (p < 0.05), and the phenotype was rescued by FST-treatment in the HFD animals. Similarly, reduced cold plate responsiveness was also observed in HFD animals, which was rescued by FST-treatment. No differences in thermal peripheral pain were found with surgery. Additionally, FST treatment protected HFD animals from mechanical algesia at the knee post-DMM surgery (Fig. 1). Conclusions: Overexpression of FST decreased body fat, pain sensitivity, and obesity-associated systemic inflammation, while increasing whole body BMD in mice with HFD-induced obesity. These findings further support the important role of muscle in obesity and inflammation, which are well-known risk factors associated with OA onset and progression. We are currently evaluating the relationship among these factors in these mice, and whether FST treatment will mitigate OA development due to obesity and joint injury.Tabled 1Serum Markers Decreased with FST Treatment in HFD Animals compared to HFD animals with GFP.MarkerHFD-GFP Mean ± SEM (ng/mL)HFD-FST Mean ± SEM (ng/mL)p-valueCXCL1/KC0.25 ± 0.030.16 ± 0.020.050CXCL10/IP-100.09 ± 0.010.06 ± 0.040.005, #IL-1α1.03 ± 0.170.53 ± 0.080.022*IL-1β0.06 ± 0.020.03 ± 0.010.028*Leptin137.93 ± 17.4317.69 ± 4.640.001*Insulin25.22 ± 4.884.30 ± 0.650.001** indicates p < 0.05; # indicates p < 0.05 between DMM animals and control surgery animals. Open table in a new tab * indicates p < 0.05; # indicates p < 0.05 between DMM animals and control surgery animals.

Cacao Polyphenols Potentiate Anti-Platelet Effect of Endothelial Cells and Ameliorate Hypercoagulatory States Associated with Hypercholesterolemia.

Platelets are related to the formation of blood clots that play a crucial role in thrombosis and other cardiovascular diseases. Cocoa, derived from Theobroma cocoa, has been widely used as functional food for improving cardiovascular health. In the present study, the direct and indirect effects of cacao polyphenols (CPs) were investigated on human platelet aggregation associated with endothelial cells (ECs) senescence. In addition, the effect of CPs on high-fat diet- (HFD-) induced hypercoagulatory states in rats was evaluated. CPs directly inhibited the human platelet aggregation induced by thromboxane analogue, U46619, and treatment of CPs on senescent endothelial cells markedly restored inhibitory effect of ECs on platelet aggregation. Nitric oxide (NO) from ECs is known as modulator of platelet aggregation and CPs increased eNOS activity in ECs and coronary artery. In animal model, increased TG level induced by high fat diet (HFD) was significantly decreased by CPs administration. In addition, the HFD animal had shorter bleeding time, and CPs administration attenuated the HFD-induced changes. Taken together, the present study indicates that CPs have potent anti-platelet effects most likely by direct and indirect effect via ECs and have the potential for lowering the risk of cardiovascular disease-related hypercoagulation due to hypercholesterolemia.

Presence of an Inhibitory Glycinergic Current in Dorsal Motor Nucleus of the Vagus Neurons Regulating Gastric Functions in Offspring of Maternal High Fat Diet Rats

The perinatal period is critically important in the development of autonomic neurocircuits innervating the gastrointestinal (GI) tract influencing gastric emptying and appetite regulation. Maternal overnutrition during this perinatal time period has been shown to predispose offspring to obesity, possibly due to developmental alterations in autonomic neurocircuits. Previous studies suggest there is an increased inhibitory tone of gastric projecting dorsal motor nucleus of the vagus nerve (DMV) neurons of rats fed a maternal high fat diet (HFD), but the underlying mechanisms have yet to be determined. The aims of this study were to assess the effects of a perinatal HFD on (i) inhibitory synaptic inputs to gastric‐projecting DMV neurons and (ii) gastric tone and motility.Whole cell patch clamp recordings (n=6–8 per group) were made from gastric projecting DMV neurons in brainstem slices from Sprague‐Dawley rats (4–6 weeks of age) fed either a control or HFD (14 or 60%kcal from fat, respectively) from embryonic day 13. Gastric motility and tone (n=4–6 rats per group) was measured via miniature strain gauges attached to the antrum and corpus of anesthetized rats.Superfusion of the non‐selective GABAA receptor antagonist, bicuculline (50μM), induced a larger inward current in DMV neurons from maternal HFD vs control animals (44.6±7.9pA vs 21.7±1.7pA; P&lt;0.05). In contrast, inward currents of similar magnitude were evoked in maternal HFD vs control neurons in response to the phasic (synaptic) GABAA receptor antagonist, gabazine (25μM), (10.5±1.0pA vs 8.6±1.4pA); subsequent addition of bicuculline uncovered a larger tonic (extrasynaptic) GABAA current (26.2±2.0pA vs 15.3±1.2pA), suggesting a greater overall inhibition of maternal DMV neurons due to a larger tonic GABAA current. Since bicuculline may be non‐selective at higher concentrations, we investigated whether other inhibitory neurotransmitters were also playing a role. The glycine receptor antagonist, strychnine (1μM), induced an inward current in maternal HFD, but not control, DMV neurons (22.8±2.1pA vs 2.9±1.5pA). Strychnine also increased the rise time (3.8±0.3ms vs 2.8±0.2ms), decay time (6.3±0.8ms vs 4.1±0.5ms), and half width (5.9±0.7ms vs 3.9±0.4ms; P&lt;0.05 for each) of miniature inhibitory postsynaptic currents (mIPSCs) in maternal HFD neurons, but had no effect in control neurons (mIPSC rise time, 3.0±0.2ms vs 3.1±0.2ms; decay time 3.6±0.2ms vs 3.7±0.4ms; half‐width 3.5±0.2ms vs 3.6±0.4ms; P&gt;0.05 for each). Microinjection of strychnine (300pmoles/60nl) into the DMV increased gastric tone and motility in the antrum in maternal HFD rats (364±130mg increase in tone and 233±199% increase in motility) but not in control rats (29.5±29.5mg increase in tone and 13±26% increase in motility), suggesting that tonic glycinergic inputs onto DMV neurons decreases vagal efferent outflow and reduces basal gastric motility and tone in maternal HFD rats.The studies suggest that perinatal overnutrition results in the expression of a novel tonic glycinergic input onto gastric projecting DMV neurons, increasing the inhibitory drive to the stomach. Such alterations in brainstem neurocircuit development as a consequence of maternal over‐nutrition may induce gastric dysmotility and dysregulated gastointestinal reflexes, hence influencing offspring to develop obesity.Support or Funding InformationFunded by NIH 078364 and NSF IOS1148978

Nigella sativa oil extracts improves dyslipidaemia in high fat diet (HFD) animal model

Introduction: Nigella sativa has been reported to have anti-inflammatory and antioxidative properties. We aimed to investigate the effects of Nigella sativa oil extract on the high fat diet (HFD) fed animal model. Methods: Thirty male Sprague-Dawley rats were used. Four groups (n=7) were fed with high fat diet (HFD) rat pellet orally for 10 weeks. The rats with HFD groups were subsequently randomly divided again into 4 groups. One group was continued with HFD while the other 3 groups were continued with the HFD in addition to Nigella sativa oil extract treatment at three different concentrations (0.5, 1 and 1.5 ml/kg/day) for another 4 weeks. Blood biochemical analysis and histological assessment of liver were subsequently performed. Results: Animals fed with HFD had increased aspartate aminotransferase (AST), alkaline phosphatase (ALP), albumin, cholesterol/HDL ratio, triglyceride (TG) levels, but decreased globulin, albumin/globulin ratio and high density lipoprotein (HDL) levels. Meanwhile, HFD animals treated with Nigella sativa oil extract showed a significant increase in the globulin and HDL levels, but reduced ALP, albumin, cholesterol/HDL ratio and TG levels. Nigella sativa oil extract at the higher doses displayed much better effect. Histologically the liver in all groups exhibited micro vesicular steatosis. Conclusions: Nigella sativa oil extracts improve the dyslipidaemia in animal models with HFD.

Hepatic mTORC1 Opposes Impaired Insulin Action to Control Mitochondrial Metabolism in Obesity

Dysregulated mitochondrial metabolism during hepatic insulin resistance may contribute to pathophysiologies ranging from elevated glucose production to hepatocellular oxidative stress and inflammation. Given that obesity impairs insulin action but paradoxically activates mTORC1, we tested whether insulin action and mammalian target of rapamycin complex 1 (mTORC1) contribute to altered invivo hepatic mitochondrial metabolism. Loss of hepatic insulin action for 2weeks caused increased gluconeogenesis, mitochondrial anaplerosis, tricarboxylic acid (TCA) cycle oxidation, and ketogenesis. However, activation of mTORC1, induced by the loss of hepatic Tsc1, suppressed these fluxes. Only glycogen synthesis was impaired by both loss of insulin receptor and mTORC1 activation. Mice with a double knockout of the insulin receptor and Tsc1 had larger livers, hyperglycemia, severely impaired glycogen storage, and suppressed ketogenesis, as compared to those with loss of the liver insulin receptor alone. Thus, activation of hepatic mTORC1 opposes the catabolic effects of impaired insulin action under some nutritional states.

Early High-Fat Feeding Induces Alteration of Trace Element Content in Tissues of Juvenile Male Wistar Rats.

The primary objective of the current study was to assess the influence of early high-fat feeding on tissue trace element content in young male Wistar rats. Twenty weanling male Wistar rats were divided into two groups fed standard (STD) or high-fat diet (HFD) containing 10 and 31.6% of total calories from fat, respectively, for 1month. Serum lipid spectrum, apolipoproteins, glucose, insulin, adiponectin, and leptin levels were assessed. The level of trace elements was estimated using inductively coupled plasma mass spectrometry. High-fat feeding significantly increased epidydimal (EDAT) and retroperitoneal adipose tissue (RPAT), as well as total adipose tissue mass by 34, 103, and 59%, respectively. Serum leptin levels in HFD animals were twofold higher than those in the control rats. No significant difference in serum lipid spectrum, apolipoproteins, glucose, adiponectin, and insulin was detected between the groups. HFD significantly altered tissue trace element content. In particular, HFD-fed animals were characterized by significantly lower levels of Cu, I, Mn, Se, and Zn in the liver; Cr, V, Co, Cu, Fe, and I content of EDAT; Co, Cu, I, Cr, V, Fe, and Zn concentration in RPAT samples. At the same time, only serum Cu was significantly depressed in HFD-fed animals as compared to the control ones. Hair Co, Mn, Si, and V levels were significantly increased in comparison to the control values, whereas Se and I content was decreased. HFD feeding induced excessive adiposity and altered tissue trace element content in rats without insulin resistance, adiponectin deficiency, and proatherogenic state. Hypothetically, trace element disbalance may precede obesity-associated metabolic disturbances.

Proatherosclerotic Effect of the α1-Subunit of Soluble Guanylyl Cyclase by Promoting Smooth Muscle Phenotypic Switching

Soluble guanylate cyclase (sGC), a key enzyme of the nitric oxide signaling pathway, is formed as a heterodimer by various isoforms of its α and β subunit. GUCY1A3, encoding the α1 subunit, was identified as a risk gene for coronary artery disease and myocardial infarction, but its specific contribution to atherosclerosis remains unclear. This study sought to decipher the role of Gucy1a3 in atherosclerosis in mice. At age 32 weeks and after 20 weeks of standard or high-fat diet, Gucy1a3(-/-)/Ldlr(-/-) mice exhibited a significant reduction of the atherosclerotic plaque size at the aortic root and the aorta for high-fat diet animals as compared with Ldlr(-/-) control mice. Collagen content in plaques in the aortic root was reduced, suggesting an alteration of smooth muscle cell function. Proliferation and migration were reduced in Gucy1a3(-/-) primary aortic smooth muscle cells (AoSMCs), and proliferation was also reduced in human AoSMCs after inhibition of sGC by 1H-[1,2,4] oxadiazolo [4,3-a] quinoxalin-1-one. Gucy1a3 deficiency in AoSMCs prevents their phenotypic switching, as indicated by the differential expression of marker proteins. The inherited Gucy1a3(-/-) loss exerts an atheroprotective effect. We suggest that sGC activity promotes the phenotypic switching of smooth muscle cells from a contractile to a synthetic state, fostering the formation of atherosclerosis. Preventing this switch by sGC inhibition may provide a novel target in atherosclerotic disease.

Maternal High Fat Diet Increases an Inhibitory Extrasynaptic Tonic GABAA Current in Dorsal Motor Nucleus of the Vagus Neurons Regulating Gastric Functions

The perinatal period is critically important in the development of autonomic neurocircuits innervating the gastrointestinal (GI) tract influencing gastric emptying and appetite regulation. Developmental alterations may affect these GI neurocircuits possibly contributing to the energy imbalance seen in obesity. Previous studies suggest there is an increased GABAergic inhibition of gastric projecting dorsal motor nucleus of the vagus nerve (DMV) neurons in the brainstem of rats fed a high fat diet (HFD) during the perinatal period, but the underlying neural mechanisms and the effects on gastric motility have yet to be clarified. The aims of this study was to assess the effects of perinatal HFD on (i) inhibitory synaptic inputs to gastric‐projecting DMV neurons and (ii) gastric tone and motility.Whole cell patch clamp recordings were made from gastric projecting DMV neurons in thin brainstem slices from Sprague‐Dawley rats (4–6 weeks of age) fed either a control or HFD (14 or 60%kcal from fat, respectively) from embryonic day 13.In the present study, perinatal HFD rats were not significantly heavier than age‐matched control rats (6 weeks; 232±17g vs 209±5g). Superfusion with the non‐selective GABAA receptor antagonist, bicuculline (50μM), induced a larger inward current in DMV neurons from perinatal HFD vs control animals (55.7±14.1pA vs 21.7±1.7pA, n=8, P&lt;0.05), suggesting a greater overall tonic inhibition. While superfusion with the phasic (synaptic) GABAA receptor antagonist, gabazine (25μM), did not reveal any difference in the resulting inward current between perinatal HFD vs control DMV neurons (11.2±1.4pA vs 6.7±1.5pA, n=6, P&gt;0.05), the addition of bicuculline into the bath uncovered a larger tonic (extrasynaptic) GABAA current in perinatal HFD vs control DMV neurons (36.4±3.47pA vs 20.8±2.24pA, n=6, P&lt;0.05), suggesting that the greater overall inhibition of perinatal DMV neurons results from a larger tonic GABAA current.The studies imply that perinatal overnutrition increases the tonic GABAergic inhibition of gastric projecting DMV neurons via an increase in activation of extrasynaptic GABAA receptors. Such alterations in brainstem neurocircuit development as a consequence of maternal over‐nutrition may induce gastric dysmotility and dysregulated gastointestinal reflexes, hence influence offspring to develop obesity.Support or Funding InformationFunded by NIH 078364 and NSF IOS1148978

Depressive like Behavior and Changes in the Gut Microflora are Associated with a High Fat Diet in Male Sprague Dawley Rats

Over the past decade, obesity has become an increasing problem worldwide according to the World Health Organization. Due to increased workload, stress, and other obligations, people tend to eat more often outside the home, frequently picking meals with a higher content of fat, and calories (called “the western diet”) resulting in increased body mass index (BMI) and obesity. In fact, several studies suggest that obesity produces gut dysfunction, inflammation, changes in the gut microflora, and mental disorders. However, it is not yet clear whether mental disorders, such as depression, can be induced in a model of obesity.AIMTo study whether a high fat diet will cause changes in the gut microflora, which results in depression.METHODSTwo‐month‐old male Sprague Dawley rats were fed with a High Fat diet (HFD), or Low Fat Diet (LFD), for a month. After diet, both groups were evaluated for depressive and anxiety like behaviors. Feces were evaluated to ascertain the composition of the gut microflora. Samples such serum and gut were evaluated for inflammatory markers.RESULTSSignificant depressive like behavior was found in the HFD group compared with LFD rats, (p&lt;0.05; n=14) however no differences were found in anxiety like behavior. Although no differences in absolute weight were found between the two groups, molecular analysis found higher epididymis fat tissue (p&lt;0.001 n=14) and a higher expression of leptin in the blood of the HFD group compared with the LFD group (p&lt;0.05 n=6). Moreover, metagenomics analysis of the feces revealed differences at 30 days at the family level (p&lt;0.001 n=6); a higher percent of bacteriodes were found in the HFD animals compared to LFD.CONCLUSIONThese data indicate that weight alone is not an indicator of obesity, but the type of food consumption can initiate changes in the gut microflora and blood, which may be associated with behavioral changes such as depressionSupport or Funding InformationRCMIG12 MD007579, GM103475‐12

A Mixture of Ethanol Extracts of Persimmon Leaf and Citrus junos Sieb Improves Blood Coagulation Parameters and Ameliorates Lipid Metabolism Disturbances Caused by Diet-Induced Obesity in C57BL/6J Mice.

This study investigated the effects of a flavonoid-rich ethanol extract of persimmon leaf (PL), an ethanol extract of Citrus junos Sieb (CJS), and a PL-CJS mixture (MPC) on mice fed a highfat diet (HFD). We sought to elucidate the mechanisms of biological activity of these substances using measurements of blood coagulation indices and lipid metabolism parameters. C57BL/6J mice were fed a HFD with PL (0.5% (w/w)), CJS (0.1% (w/w)), or MPC (PL 0.5%, CJS 0.1% (w/w)) for 10 weeks. In comparison with data obtained for mice in the untreated HFD group, consumption of MPC remarkably prolonged the activated partial thromboplastin time (aPTT) and prothrombin time (PT), whereas exposure to PL prolonged aPTT only. Lower levels of plasma total cholesterol, hepatic cholesterol, and erythrocyte thiobarbituric acid-reactive substances, hepatic HMG-CoA reductase, and decreased SREBP-1c gene expression were observed in mice that received PL and MPC supplements compared with the respective values detected in the untreated HFD animals. Our results indicate that PL and MPC may have beneficial effects on blood circulation and lipid metabolism in obese mice.

Acute Injury in Natural Diet-Induced Fatty Livers - A Model for Therapy Development

Given the diabesity and metabolic syndrome epidemics, fatty liver disease is reaching epidemic proportions. Relatively indolent, this disease is often asymptomatic and the patient is often made aware of its presence only during a routine physical exam. Nevertheless, fatty livers are more susceptible to insult compared to their non-fatty counterparts and persons with fatty livers are at increased risk for morbidity and mortality following consumption of commonly used substances such as alcohol (EtOH) and acetaminophen (APAP). We have developed a rat model of natural diet-induced fatty liver disease and evaluated the effects of two commonly used substances viz. EtOH and APAP in this phenotype. High fat diet (HFD) fed animals exhibited steatosis, liver inflammation and liver fibrosis with an increase in serum aspartate aminotransferase. Bolus administration of EtOH, which was without effect on the livers from standard diet fed animals, had a profound and adverse impact on the HFD fatty liver. Similarly, APAP administration which was without effect on liver function tests in control animals, also provoked an increase in liver enzymes in HFD animals. Treatment with the poly(ADP-ribose) polymerase-1 inhibitor (PARP-1), veliparib, reduced the increase in liver function tests secondary to EtOH and APAP. This model forms the framework for identification of fatty liver disease biomarkers given that this disease is relatively asymptomatic but fraught with risk for acute injury. This model also forms the framework for evaluation of novel drugs for acute injury in fatty livers especially given that current strategies for management of acute liver failure in non-fatty livers are inadequate. Also, relevant patents related to the use of liver biomarkers as diagnostic are discussed. Keywords: Acetaminophen, acute, alcohol, fibrosis, injury, liver, steatosis.

Reduction of CTRP9, a novel anti-platelet adipokine, contributes to abnormal platelet activity in diabetic animals.

Platelet hyper-reactivity is a crucial cause of accelerated atherosclerosis increasing risk of thrombotic vascular events in diabetic patients. The mechanisms leading to abnormal platelet activity during diabetes are complex and not fully defined. The current study attempted to clarify the role of CTRP9, a novel adiponectin paralog, in enhanced platelet activity and determined whether CTRP9 may inhibit platelet activity. Adult male C57BL/6 J mice were randomized to receive high-fat diet (HFD) or normal diet (ND). 8 weeks after HFD, animals were sacrificed, and both plasma CTRP9 and platelet aggregation were determined. HFD-fed animals increased weight gain significantly, and became hyperglycemic and hyperinsulinemic 8 weeks post-HFD. Compared to ND animals, HFD animals exhibited significantly decreased plasma CTRP9 concentration and increased platelet response to ADP, evidenced by augmented aggregation amplitude, steeper aggregation slope, larger area under the curve, and shorter lag time (P < 0.01). A significant negative correlation between plasma CTRP9 concentration and platelet aggregation amplitude was observed. More importantly, in vitro pre-treatment with CTRP9 significantly inhibited ADP-stimulated platelet activation in platelet samples from both ND and HFD animals. Taken together, our results suggest reduced plasma CTRP9 concentration during diabetes plays a causative role in platelet hyper-activity, contributing to platelet-induced cardiovascular damage during this pathologic condition. Enhancing CTRP9 production and/or exogenous supplementation of CTRP9 may protect against diabetic cardiovascular injury via inhibition of abnormal platelet activity.

Administration of granulocyte-colony stimulating factor accompanied with a balanced diet improves cardiac function alterations induced by high fat diet in mice.

Background/ObjectivesHigh fat diet (HFD) is a major contributor to the development of obesity and cardiovascular diseases due to the induction of cardiac structural and hemodynamic abnormalities. We used a model of diabetic cardiomyopathy in C57Bl/6 mice fed with a HFD to investigate the effects of granulocyte-colony stimulating factor (G-CSF), a cytokine known for its beneficial effects in the heart, on cardiac anatomical and functional abnormalities associated with obesity and type 2 diabetes.MethodsGroups of C57Bl/6 mice were fed with standard diet (n = 8) or HFD (n = 16). After 36 weeks, HFD animals were divided into a group treated with G-CSF + standard diet (n = 8) and a vehicle control group + standard diet (n = 8). Cardiac structure and function were assessed by electrocardiography, echocardiography and treadmill tests, in addition to the evaluation of body weight, fasting glicemia, insulin and glucose tolerance at different time points. Histological analyses were performed in the heart tissue.ResultsHFD consumption induced metabolic alterations characteristic of type 2 diabetes and obesity, as well as cardiac fibrosis and reduced exercise capacity. Upon returning to a standard diet, obese mice body weight returned to non-obese levels. G-CSF administration accelerated the reduction in of body weight in obese mice. Additionally, G-CSF treatment reduced insulin levels, diminished heart fibrosis, increased exercise capacity and reversed cardiac alterations, including bradycardia, elevated QRS amplitude, augmented P amplitude, increased septal wall thickness, left ventricular posterior thickening and cardiac output reduction.ConclusionOur results indicate that G-CSF administration caused beneficial effects on obesity-associated cardiac impairment.

Effects of vitamin d3 on ppar-α and its target gene nfkbia on a rat model of high carbohydrate high fat diet induced non alcoholic steatohepatitis

Accumulating data suggesting that Vitamin D3 could be of a potential therapeutic value for non-alcoholic fatty liver disease (NAFLD) ,but its underlying mechanisms are still under research .Vitamin D3 up regulates PPAR-α and downstream gene nuclear factor-kappa-B inhibitor subunit-alpha (NFKBIA) which may represent a novel strategy for the management of NASH. Therefore, in this study, we investigated the possible protective role of vitamin D3 on high carbohydrate high fat diet (HCHFD) animal model of NASH through PPAR-α and NFKBIA pathway Methods: Thirty male Sprague-Dawley rats were equally divided into three groups; group I: (negative control group) fed standard rat chow diet for 8 weeks. group II: received HCHFD for 8 weeks. group III: fed as group II for 8 weeks and received also, I.P administration of vitamin D3 at a dose of 5 μg/kg twice weekly from the fourth week till the end of the study. At the end of the experiment, serum triglycerides, total cholesterol, liver enzymes, serum albumin, serum bilirubin and liver tissue oxidative stress markers (MDA, SOD and GSH) were measured. Hepatic histopathological alterations were assessed by hematoxylin and eosin, Masson’s trichrome, and Oil Red O staining ; the expressions of NF-κ B p65 (ReIA) was estimated in liver tissues using immunohistochemistry and the expression of hepatic PPAR-α and NFKBIA genes were estimated at the level of mRNA by quantitative real-time PCR. Results: Nonalcoholic steatohepatitis (NASH) model was confirmed by histopathological examination as indicated by elevated NAFLD activity score (NAS) as well as elevation of serum liver enzymes , hyperlipidemia in the form elevated plasma triglycerides and total cholesterol, oxidative stress state (high MDA, low GSH and SOD) and down regulation of PPAR-α and NFKBIA mRNA in liver tissues which was concomitant with up regulation of inflammatory marker NF-κ B-p65(ReIA) . On the other hand, vitamin D3 ameliorated NASH by attenuation of oxidative stress (low MDA, high GSH and SOD) and up regulation of PPAR-α and NFKBIA genes at the level of mRNA in the liver tissues which was accompanied by decrease in hepatocyte nuclear NF-κ B-p65 (ReIA) immunohistochemical brownish staining. Conclusion: Vitamin D3 attenuates NASH in HCHFD rat model by attenuation of redox state and up regulation of PPAR-α and its downstream gene NFKBIA in liver tissues.

Abstract 14092: Reduction of Atherosclerosis in Gucy1a3 Deficient Mice

Introduction: GUCY1A3 encodes the α1 subunit of the soluble guanylate cyclase (sGC) and has been associated with CAD/MI by genome-wide association studies (GWAS) and in a rare extended family with MI. The α1 and the β1 subunit form the heterodimeric sGC, the nitric oxide (NO) receptor. NO plays an important role in the human cardiovascular system. The aim of our study is to define the role of GUCY1A3 in atherosclerosis using a mouse atherogenic model. Methods: Gucy1a3 KO mice were backcrossed on an Ldlr KO atherogenic background. Four groups were used, i.e. C57BL/6 (WT), Gucy1a3 KO, Ldlr KO and Gucy1a3-Ldlr double KO. All groups were fed high fat diet (HFD) or standard diet (SD) starting at the age of 12-14w and for 20 weeks. Blood pressure was studied at the end of the experiment in the HFD. At the start and end of the diet, lipid metabolism parameters (TC, HDLC, LDLC and TG) were analyzed. Plaque lesion size and collagen and macrophages content were studied. As smooth muscle cells play a key role in atherosclerosis, primary mouse aortic smooth muscle cells (maSMC) were isolated from WT and Gucy1a3 KO animals and subjected to proliferation and migration assays. Results: Blood pressure was studied in HFD animals, showing, as expected, an increase of systolic and diastolic blood pressure in double KO animals when compared to Ldlr KO animals. Regarding lipid levels, no differences were found in the double KO to Ldlr KO comparison under HFD, but a reduction in total cholesterol was found in double KO under SD. For atherosclerosis, a significant reduction in plaque area at the aortic root of double KO when compared to the Ldlr KO animals was demonstrated in both SD and HFD. The reduction was further confirmed in an additional independent experiment under SD. Similarly, a significant reduction of plaque lesion was found in the whole aorta of double KO when compared to the Ldlr KO under HFD. Moreover, a reduction in collagen content was found in double KO mice under HFD when compared to Ldlr KO animals. The migration and proliferation assays show a significant reduction in both migration and proliferation of the Gucy1a3 KO maSMC compared to WT. Conclusions: Our results point to an atherogenic role of Gucy1a3 in mice probably induced via smooth muscle migration and proliferation defects.

The effect of high-fat diet on rat's mood, feeding behavior and response to stress.

An association between obesity and depression has been indicated in studies addressing common physical (metabolic) and psychological (anxiety, low self-esteem) outcomes. Of consideration in both obesity and depression are chronic mild stressors to which individuals are exposed to on a daily basis. However, the response to stress is remarkably variable depending on numerous factors, such as the physical health and the mental state at the time of exposure. Here a chronic mild stress (CMS) protocol was used to assess the effect of high-fat diet (HFD)-induced obesity on response to stress in a rat model. In addition to the development of metabolic complications, such as glucose intolerance, diet-induced obesity caused behavioral alterations. Specifically, animals fed on HFD displayed depressive- and anxious-like behaviors that were only present in the normal diet (ND) group upon exposure to CMS. Of notice, these mood impairments were not further aggravated when the HFD animals were exposed to CMS, which suggest a ceiling effect. Moreover, although there was a sudden drop of food consumption in the first 3 weeks of the CMS protocol in both ND and HFD groups, only the CMS-HFD displayed an overall noticeable decrease in total food intake during the 6 weeks of the CMS protocol. Altogether, the study suggests that HFD impacts on the response to CMS, which should be considered when addressing the consequences of obesity in behavior.