Neurobehavioural Deficits, Atherogenic Risk, and Oxido-inflammatory Response in Mice Fed with Combined Sodium Benzoate and Monosodium Glutamate Supplementation in Normal and High-fat Diets

The effect of lycopene supplementation on the antioxidant system was investigated by analyzing lipid peroxide levels, glutathione contents, and antioxidant enzyme activities in Mongolian gerbils fed a high fat diet. Gerbils were fed on each experimental diet for 6 weeks; normal diet (NC), normal diet with 0.05% lycopene (NL), high fat diet (HF), and a high fat diet with 0.05% lycopene (HFL). Dietary supplementation of lycopene increased hepatic lycopene level in gerbils fed a normal or high fat diet (P < 0.05). Liver and erythrocyte concentrations of lipid peroxide increased in gerbils fed a high fat diet, whereas lycopene supplementation decreased liver and erythrocyte concentrations of lipid peroxide (P < 0.05). Hepatic total glutathione content was higher in the NL group than that in the NC group (P < 0.05). Total antioxidant status in plasma increased following lycopene supplementation compared with that of the non-lycopene supplemented groups (P < 0.05). Hepatic catalase activity increased following dietary lycopene supplementation (P < 0.05). Superoxide dismutase activity in liver remained unchanged with lycopene supplementation, but erythrocyte superoxide dismutase activity increased in NL group compared with NC group (P < 0.05). Glutathione-S-transferase activity increased in the NL group compared to NC group (P < 0.05). Liver and erythrocyte glutathione peroxidase activity increased significantly in the NL group compared to that in the HF group (P < 0.05). Liver glutathione reductase activity was higher in the NL group than that in the NC group (P < 0.05). These results suggest that lycopene supplementation may be efficient for preventing chronic diseases induced by oxidative stress related to high fat diet.

Dietary supplementation with monosodium l-glutamate modifies lipid composition and gene expression related to lipid metabolism in growing pigs fed a normal- or high-fat diet

Obesity is a risk factor for various diseases, including cardiovascular disease, diabetes, renal disease, hypertension, cancer, and neural disease. Adipose tissue in animals is important for the mobilization of lipids, milk production, deposition of fat in different depots, and muscle and meat production. Understanding the genetic and physiological causes of metabolic disease is a priority in biomedical genome research. In this study, we examined several variables in mice fed a high-fat diet, including serum composition, body weight, total calorie intake, and differentially expressed genes. Body weight and blood glucose levels were not significantly different between animals fed high-fat and normal diets. However, high-fat diet groups showed reduced calorie and food intakes. Levels of sodium, ionized calcium, glucose, hematocrit, hemoglobin, pH, PCO2, PO2, TCO2+, HCO3+, base excess, and SO2 in the blood were not significantly different between mice fed high-fat and normal diets. Serum potassium concentration, however, was lower in mice a high-fat diet. Differentially expressed genes were also compared between the two groups. The purpose of this study was to discover new genes as a result of annealing control primer (ACP) PCR using 20 random primers. Five down regulated genes were identified and three of others were up-regulated by high-fat diet. Known genes were excluded from this result. In addition, the relationships among candidate genes and high-fat diet should be investigated according to potassium concentration in the blood. In conclusion, mice fed normal and high-fat diets showed no significant difference in body weight, whereas high-fat diet led to changes in blood composition and differential expression of several genes. These findings may provide a better understanding of the mechanisms underlying the association between obesity and metabolic diseases.

Aloe vera gel supercritical CO2 extract (AVGE) has been shown to contain five phytosterols, reduce visceral fat accumulation, and influence the metabolism of glucose and lipids in animal model experiments. Recent epidemiologic studies have shown that obesity is an established risk factor for several cancers including colorectal cancer. Therefore, we examined the effects of AVGE on intestinal polyp formation in Apc-deficient Min mice fed a high-fat diet. Male Min mice were divided into normal diet (ND), high fat diet (HFD), low dose AVGE (HFD+LAVGE) and high dose AVGE (HFD+HAVGE) groups. The ND group received AIN-93G diet and the latter 3 groups were given modified high-fat AIN-93G diet (HFD) for 7 weeks. AVGE was suspended in 0.5% carboxymethyl cellulose (CMC) and administered orally to mice in HFD+LAVGE and HFD+HAVGE groups every day (except on Sunday) for 7 weeks at a dose of 3.75 and 12.5 mg/kg body weight, respectively. ND and HFD groups received 0.5% CMC alone. Between weeks 4 and 7, body weights in the HFD and HFD+LAVGE groups were reduced more than those in the ND group. However, body weights were not reduced in the HFD+HAVGE group. Mice were sacrificed at the end of the experiment and their intestines were scored for polyps. No significant differences were observed in either the incidence and multiplicity of intestinal polyps (≥0.5 mm in a diameter) among the three groups fed HFD. However, when intestinal polyps were categorized by their size into 0.5-1.4, 1.5-2.4, or ≥2.5 mm, the incidence and multiplicity of large polyps (≥2.5 mm) in the intestine in the HFD+HAVGE group were significantly lower than those in the HFD group. We measured plasma lipid (triglycerides and total cholesterol) and adipocytokine [interleukin-6 and high molecular weight (HMW) adiponectin] levels as possible indicators of mechanisms of inhibition. The results showed that HMW adiponectin levels in the HFD group were significantly lower than those in the ND group. However, the levels in the HFD+HAVGE group were significantly higher than those in the HFD group. These results indicate that HAVGE reduced large-sized intestinal polyps and ameliorated reduction in plasma HMW adiponectin levels in Min mice fed HFD.

Endothelin-1 (ET-1) is a potent vasoconstrictor and proinflammatory protein that is increased in adipose tissue of high fat diet (HFD) fed mice and contributes to cardiovascular and metabolic disease risk in obesity. Obesity causes adipose tissue hypoxia followed by infiltration of pro-inflammatory T cells and macrophages, adipocyte dysfunction (adipokine dysregulation and inability to store excess energy), and eventual peripheral insulin resistance. Our lab recently reported that adipocyte knockout of the ETB receptor (adETBKO) led to robust attenuation of adipose tissue inflammation, reduction in circulating adiponectin, and increased insulin sensitivity in mice fed HFD. Therefore, we hypothesized that treatment with macitentan, a dual ETA/ETB receptor antagonist, will reduce cardio-metabolic disease risk similar to what is observed in obese adETBKO mice. To test this hypothesis, C57BL/6J mice were fed either normal diet (NMD) or HFD for 8 weeks followed by 2 weeks of treatment with either vehicle or macitentan (30mg/kg/day) while diet was maintained. HFD mice had significantly higher fat mass than NMD mice, with no significant effect of treatment with macitentan. In addition, hypoxia inducible factor alpha ( Hif-1α ) mRNA, which increases as oxygen tension decreases, was elevated in visceral adipose of HFD fed mice compared to NMD (30963±3755 vs. 49771±5954 copies per 50 ng RNA; NMD vs. HFD; p&lt;0.05 by two-way ANOVA). Macitentan reduced Hif-1α transcripts in NMD fed mice and attenuated the increase in HFD fed mice (25050±2982 vs. 32353±5061 copies per 50 ng RNA; NMD vs. HFD). Likewise, HFD fed mice had a significant increase in the proinflammatory cytokine tumor necrosis factor α mRNA (241±50.1 vs. 792.3±120.4 copies per 50 ng RNA; NMD vs. HFD; p&lt;0.0001) in visceral adipose that was attenuated in mice treated with macitentan (209±29 vs. 431±44 copies per 50 ng RNA; NMD macitentan treated vs. HFD macitentan treated). Further, mRNA copies of the cytokines interleukin (IL) 10 and 12b were increased in visceral adipose of mice fed HFD compared to NMD (102±21 NMD vs. 283±58 HFD and 16±4 NMD vs 106±28 HFD copies per 50 ng RNA; IL10 and IL12b, respectively; both p&lt;0.05 by two-way ANOVA), and this increase was attenuated by treatment with macitentan, (90±14 NMD vs. 170±37 HFD and 16±3 NMD vs. 55±14 HFD copies per 50 ng RNA; IL10 and IL12b, respectively). As expected, adiponectin mRNA was significantly reduced in HFD fed mice compared to NMD, however, there was no detectable difference in macitentan treated mice, suggesting that ETB receptor inhibition was not achieved at the adipocytes. Taken together, these data suggest that ET-1 receptor blockade with macitentan improves inflammation in visceral adipose tissue of obese mice. We predict that this is achieved in part by increasing oxygen delivery to the adipose tissue, but not through direct inhibition of ETB receptors on adipocytes. This work was funded by NIH grants R25 HL121042, P20 GM104357, R01 DK124327, and T32 HL105324. This abstract was presented at the American Physiology Summit 2025 and is only available in HTML format. There is no downloadable file or PDF version. The Physiology editorial board was not involved in the peer review process.

The effect of a high fat, low carbohydrate, low protein diet on the in vivo oxidation of 17 beta-estradiol was studied using radiometric methods. Five male chimpanzees were fed a normal (13%) fat diet or a high (65%) fat diet for 8 weeks. After a 4-week rest period, the animals were fed the alternative diet. The mean percent oxidation of 16 alpha-[3H]estradiol-17 beta 24 h after injection was 3.8 +/- 1.3% (+/- SD) on the normal diet vs. 18.4 +/- 4.7% on the high fat diet (P less than 0.01). In contrast, the mean percent oxidation of 2-[3H]estradiol 24 h after injection was 31.6 +/- 3.8% (+/- SD) on the normal diet vs. 20.0 +/- 3.5% on the high fat diet (P less than 0.05). These results suggest that the oxidation of 17 beta-estradiol to estriols relative to that to catechol estrogens is increased by a high fat diet.

Free feeding (FF) with a high fat diet (HFD) causes excessive body weight gain, whereas restricted feeding (RF) with a HFD attenuates body weight gain. The effects of timing of feeding with a HFD (day vs. night) and feeding duration on energy homeostasis have not yet been investigated. In this study, we fed mice a HFD or a normal diet (ND) twice a day, during their active and inactive periods, on a schedule. The amount of food was regulated by feeding duration (2, 4 or 8 h). First, we investigated the effects of 4-h RF during active–inactive periods (ND–ND, HFD–HFD, ND–HFD or HFD–ND). Among all the 4-h RF groups, mice consumed almost the same amount of calories as those in the FF[ND] group, even those fed a HFD. Body weight and visceral fat in these three groups were lower than that in the FF[HFD] group. Second, we investigated the effects of RF duration. Body weight and visceral fat were higher in the 8-h groups than in the 4-h groups. Body weight and visceral fat were higher in the 2-h groups than in the 4-h groups even though the 2-h groups had less food. Third, we investigated the effects of eating a HFD during the inactive period, when RF duration was extended (2, 6 or 12 h). Mice were fed with a HFD during the inactive period for 2 h and fed with a ND during the active period for 2, 6 or 12 h. Body weight and visceral fat in these mice were comparable to those in the FF[ND] mice. The results of our first set of experiments suggest that 4-h RF was an adequate feeding duration to control the effect of a HFD on obesity. The results of our second set of experiments suggest 2-h RF (such as speed-eating) and 8-h RF, representative of eating disorders, are unhealthy feeding patterns related to obesity. The results of our third set of experiments suggest that eating a HFD for a short period during the night does not affect body weight and visceral fat. Taken together, these results indicate that consideration to feeding with a HFD during the inactive period and restricting eating habits relieve the risks of body weight gain and visceral fat accumulation.

The only reliable factor that reduces the risk of colorectal carcinogenesis is physical activity. However, the underlying mechanisms remain unclear. In this study, we examined the effects of physical activity against gut microbiota, including mucosa-associated microbiota (MAM) on azoxymethane-induced colorectal tumors in obese mice. We divided the subjects into four groups: normal diet (ND), high-fat diet (HFD), ND + exercise (Ex), and HFD + Ex groups. The Ex group performed treadmill exercise for 20 weeks. Thereafter, fecal and colonic mucus samples were extracted for microbiota analysis. DNA was collected from feces and colonic mucosa, and V3-V4 amplicon sequencing analysis of the 16SrRNA gene was performed using MiSeq. The HFD group had significantly more colonic polyps than the ND group (ND 6.5 ± 1.3, HFD 11.4 ± 1.5, p < 0.001), and the addition of Ex suppressed the number of colonic polyps in ND and HFD groups (ND 6.5 ± 1.3, ND + Ex 2.8 ± 2.5, p < 0.05). The HFD group showed significantly lower concentrations of succinic, acetic, butyric, and propionic acids (mg/g) in feces, compared with the ND group (succinic acid HFD 0.59, ND 0.17; acetic acid HFD 0.63, ND 2.41; propionic acid HFD 0.10, ND 0.47; and N-butyric acid HFD 0.31, ND 0.93). In the case of ND, succinic acid and butyric acid tended to decrease with Ex (succinic acid ND 0.17, ND + Ex 0.12; N-butyric acid ND 0.93, ND + Ex 0.74 0.74). Succinic acid, acetic acid, butyric acid, and propionic acid levels in feces were significantly lower in the HFD group than in the ND group; in both feces and mucus samples, Butyricicoccus and Lactobacillus levels were significantly lower in the HFD group. Akkermansia was significantly increased in ND + Ex and HFD + Ex groups. Diet and exercise affected the number of colorectal tumors. Furthermore, diet and exercise alter intestinal MAM, which may be involved in colorectal tumor development.

BackgroundPineapple peel is a waste component of pineapple with valuable source of metabolites as phytoactive compounds in ameliorating metabolic-related disorders. This study investigated the atheroprotective and neuroprotective effects of peel extract of Ananas comosus fruit (PEAC) in normal diet (ND) and high-fat diet (HFD) fed rats. MethodsMale Wistar rats were fed ND or HFD for 9 weeks, and beginning from the 6th week animals were also orally treated with PEAC (200 mg/kg). Memory performance was assessed using Y-maze test (YMT) and novel object recognition test (NORT) while anxiolytic-like effect was assessed on the elevated plus maze (EPM). Serum cholesterol, triglycerides and HDL-C were determined, while LDL-C and atherogenic risk calculated. Serum and brain tissue malondialdehyde, reduced glutathione, catalase were determined. Brain acetylcholinesterase activity and interleukin-6 level were also determined. ResultsPEAC significantly attenuated HFD-induced reduction in correct alternation in YMT, and discrimination index in NORT. Also, PEAC demonstrated anxiolytic-like activity in EPM test. PEAC significantly improved lipid profile and decreased risk of atherogenicity in ND and HFD-fed rats. In addition, PEAC improves serum and brain antioxidant status by decreasing malondialdehyde and increasing GSH and catalase. PEAC significantly impaired HFD-induced brain acetylcholinesterase activity and IL-6 levels. ConclusionThese findings suggest that peel extract of Ananas comosus fruit may protect against diet-induced behavioral disturbances via atheroprotective, antioxidants and anti-inflammatory activities.

Maternal diet impacts in vitro culture requirements for optimal embryo development

We previously reported that acute necrotizing pancreatitis (ANP) after normal or high-fat diet is associated with a decreased number of Paneth cells in ileal crypts. Here, we ablated Paneth cells in a rat model of ANP after normal and high-fat diet to investigate the effects on disease symptoms. Adult male Sprague-Dawley rats received standard rat chow or a high-fat diet for 2 weeks, after which they were treated with dithizone to deplete Paneth cells. Six hours later, ANP was established by retrograde injection of sodium taurocholate into the biliopancreatic duct. Rats were sacrificed at 6, 12, and 24 h for assessment. We found dithizone aggravated ANP-associated pathological injuries to the pancreas and ileum in rats on high-fat or standard diets. Lysozyme expression in ileal crypts was decreased, while serum inflammatory cytokines (TNFα, IL-1β, and IL-17A) and intestinal permeability (serum DAO activity and D-lactate) were increased. Expression of tight junction proteins (claudin-1, zo-1, and occludin) was decreased. Using high-throughput 16S rRNA sequencing, we found dithizone reduced microbiota diversity and altered microbiota composition in rats on high-fat or standard diets. Dithizone decreased fecal short-chain fatty acids (SCFAs) in rats on high-fat or standard diets. Changes in intestinal microbiota correlated significantly with SCFAs, lysozyme, DAO activity, D-lactate, inflammatory cytokines, and pathological injury to the pancreas and ileum in rats on high-fat or standard diets. In conclusion, ablation of Paneth cells exacerbates pancreatic and intestinal injuries in ANP after normal and high-fat diet. These symptoms may be related to changes in the intestinal microbiota.

Objective: To investigate the changes in thyroid morphology and function in mice with thyroid-specific knockout of the clock gene Bmal1 under high-fat diet (HFD), and to examine its effects on glycolipid metabolism in mice. Methods: Construct a mouse model with specific knockout of the Bmal1 gene in the thyroid (T-Bmal1-/-) (knockout group, n=10), and use Bmal1flox/flox mice without thyroid peroxidase-cyclization recombination enzyme (T-Bmal1+/+) as the control group (non-knockout group, n=10). The mice were fed until 6 weeks (body weight 20-23 g), and then use the random number table method to evenly divide each group of mice into two subgroups. They were then fed either with normal diet (ND) or (HFD), resulting in four final groups: ND non-knockout group, HFD non-knockout group, ND knockout group, and HFD knockout group, with 5 mice in each group. From the 6th week onwards, the body weights of the mice were measured for 10 consecutive weeks. On the first day of the 14th week, the intraperitoneal glucose tolerance test (IPGTT) was conducted, and on the first day of the 15th week, the intraperitoneal insulin tolerance test (IPITT) was performed. On the first day of the 16th week after the mice were born, blood samples (0.2 ml) were taken from the eyes under anesthesia at 7∶00, 13:00, 19:00, and 1∶00 the next day, and the thyroid, liver, kidneys, spleen, inguinal white adipose tissue, and scapular brown adipose tissue were rapidly removed. Thyroid tissue morphology, thyroid function indexes, thyroid clock genes and thyroid hormone synthesis and secretion related genes expression, body weight, IPGTT and IPITT area under the curve (AUC) and lipid levels were detected and compared among all groups. Results: There was no significant difference in the number of thyroid follicles, the area of thyroid follicles and the height of thyroid follicle cavity between ND knockout group and ND non-knockout group (all P>0.05), but the height of thyroid follicle cavity in HFD knockout group was lower than that in HFD knockout group [(25.8±1.6) vs (54.4±9.5) μm, P=0.002]. The level of serum T4 in knockout group after ND or HFD feeding was higher than that in non-knockout group (all P<0.05). There was no significant difference in the mRNA expression level of sodium iodine transporter between ND knockout group and ND non-knockout group (P=0.550), but the mRNA expression level of sodium iodine transporter in HFD knockout group was higher than that in HFD non-knockout group [0.67±0.27 vs 0.20±0.09, P=0.006].There was no significant difference in weight gain and groin white fat weight between ND knockout group and ND non-knockout group (both P>0.05), but weight gain and groin white fat tissue weight in HFD knockout group were lower than those in HFD knockout group (both P<0.05). The IPGTT results showed no statistically significant difference in AUC between the ND knockout and ND non-knockout groups (P=0.226), but the HFD knockout group had a smaller AUC than the HFD non-knockout group (P=0.008). The IPITT revealed that the AUC of the ND knockout group was smaller than that of the ND non-knockout group (P=0.047). Lipid profiles were similar between the ND knockout and ND non-knockout groups (all P>0.05), but the HFD knockout group had lower levels of total cholesterol and high-density lipoprotein cholesterol than the HFD non-knockout group (both P<0.05). Conclusion: Thyroid-specific knockout of the clock gene Bmal1 can affect thyroid hormone levels and glycolipid metabolism in mice, with more pronounced effects after HFD feeding.

Loss of ceramide synthase 5 reduces the development of aortic valve stenosis in mice with high fat diet

Hematological inflammatory indices are currently suggested to assess systemic inflammation. This study aims to investigate a vitamin D supplementation effect on hematological indices of inflammation in rats. Forty-eight middle-aged male rats were allocated into a normal diet (ND) group (10% fat) and a high-fat diet (HFD) group (60% fat). The animals were fed for 26 weeks. After this period, each group was randomly divided into three subgroups, each of 8 rats: Group (1): animals were fed the ND and HFD containing 1 IU/g vitamin D for 4 months, group (2): animals were fed the ND and HFD containing 6 IU/g vitamin D for 4 months and group (3): animals were euthanized to evaluate the HFD effect. Serum 25-hydroxyvitamin D level, white blood cell count (WBCs), platelet count, platelet crit (PCT), mean platelet volume (MPV), platelet distribution width (PDW), platelet-to-lymphocyte ratio (PLR), neutrophil-to-lymphocyte ratio (NLR), monocyte-to-lymphocyte ratio (MLR) were measured. The HFD, significantly increased body weight, PCT, PDW, PLR, NLR, and MLR and significantly reduced serum vitamin D levels compared to the ND (P < 0.05). There was a significant decrease in food intake, MPV, PDW, and NLR after vitamin D supplementation in the ND-fed group (P < 0.05). A significant reduction in platelet count, PCT, and MLR was observed after vitamin D supplementation in HFD-fed rats (P < 0.05). In our study, some hemogram-derived inflammatory indices were higher in the HFD-fed group, and vitamin D supplementation lowering effects on some hematological indices were seen in both ND and HFD groups.

INTRODUCTION: While obesity is an established independent prognostic factor for breast cancer patients, the underlying mechanisms how obesity promotes breast cancer progression are not well understood. Sphingosine-1-phosphate (S1P) is a pleiotropic bioactive lipid mediator produced by sphingosine kinases (SphKs) that plays critical roles in inflammation and cancer progression. Our hypothesis is that obesity increases levels of S1P in both tumor and its microenvironment, which promotes obesity-induced inflammation and breast cancer progression. The aim of this study is to test the hypothesis. METHODS: A high fat diet (HFD) was fed C57Bl/6 mice, which is the most commonly used murine diet-induced obesity model. E0771 cells derived from C57Bl/6 mice were implanted into mammary fat pads of mice that were fed with HFD or normal diet (ND) for 12 weeks prior to the implantation. Western blot, immunofluorescent staining, RT-qPCR and LC-ESI-MS/MS assays were used. RESULTS: Mice fed with HFD for 20 weeks developed severe obesity with an almost 2 fold increase of body weight compared with ND fed mice. E0771 mouse breast cancer cells were implanted into mammary fat pad of C57Bl/6 mice, and the mice fed with HFD developed significantly larger tumors within 30 days than those fed with ND. Expression of pro-inflammatory cytokines, IL-6 and TNF-α, was increased in the tumors of HFD fed mice, compared to those fed with ND. Furthermore, immunofluorescent analysis with anti-F4/80 antibody showed that tumors from HFD fed animals recruited significantly more tumor associated macrophages than those from ND fed animals. Expression of SphK1 and S1PR1, but not SphK2, was increased in the tumors from mice fed HFD. Mass spectrometry analysis revealed that while S1P levels in the normal breast mammary fat pad were increased with HFD feeding, S1P levels were even higher in breast tumors. Consistent with increased SphK1 and S1P in tumors, S1P was also significantly increased in the tumor interstitial fluid, which is a component of the tumor microenvironment and bathes cancer cells in the tumor. S1P levels were also increased in the serum of the tumor-implanted animals fed with HFD compared with those fed with ND, while minimal changes in S1P were evident in the serum of non-tumor bearing mice fed with HFD. Furthermore, S1P levels in the lungs of tumor-implanted animals fed with HFD were significantly higher than those fed with ND. CONCLUSION: S1P is increased not only in tumor, but also in tumor microenvironment such as tumor interstitial fluid by obesity. Our results suggest that S1P has a role in obesity-induced inflammation and the cancer progression. This work was supported by the Japan Society for the Promotion of Science (JSPS) Grant-in-Aid for Scientific Research Grant Number 15H05676 and 15K15471 for M.N and 15H04927 for W.T. M.N. is supported by the Uehara Memorial Foundation, Nakayama Cancer Research Institute, and Tsukada Medical Foundation. K.T. is supported by NIH/NCI grant R01CA160688 and Susan G. Komen Investigator Initiated Research Grant IIR12222224. Citation Format: Tsuchida J, Nagahashi M, Moro K, Tatsuda K, Koyama Y, Takabe K, Wakai T. Obesity increases the lipid mediator, sphingosine-1-phosphate in the tumor and tumor microenvironment, and promotes tumor progression. [abstract]. In: Proceedings of the Thirty-Eighth Annual CTRC-AACR San Antonio Breast Cancer Symposium: 2015 Dec 8-12; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2016;76(4 Suppl):Abstract nr P1-03-03.