To explore the effect of high fat diet on proteome in mice stomachs, we constructed a model in which the mice were fed with high fat diet as the high fat diet (HFD) group or normal diet as the control (CTRL) group for 110 days. The stomachs were collected and divided into three regions (forestomach (F), corpus (C) and antrum (A)) for protein extraction and mass spectrometry analysis. Of all 9 307 identified proteins in two groups, 4 066 proteins (HFD: 3 832, CTRL: 3 654) were strictly identified by at least one unique peptide and identified twice in three replicates. Using gene ontology (GO) and interaction network analysis we analyzed differentially expressed proteins (fold change≥2) in two groups or between regions. In the whole stomach tissues, proteins up-regulated in HFD group mainly were associated with protein stabilization and protein transport. Differentially expressed proteins between regions showed that forestomach was related to the biological process of keratinization and actin assembly, while corpus and antrum mainly performed digestive function. Compared with forestomach, the corpus and antrum were more affected by the diet. Though there was no significant effect on the basic digestive function of the stomach, proteins that were involved in protein transport and lipid metabolism-related biological processes were significantly highly expressed in HFD group.
Animals ; Diet, High-Fat ; Lipid Metabolism ; Mice ; Mice, Inbred C57BL ; Protein Transport ; Proteome ; physiology ; Stomach ; physiology

Objective: To evaluate the effects of a high-protein diet on anthropometric indices and blood lipid in overweight and obese children and provide evidence for their dietary management. Methods: This was a Meta-analysis. The randomized controlled trials on the effects of a high-protein diet on anthropometric indices and blood lipid in overweight and obese children published up to January 19, 2022 were searched in PubMed, Web of Science, Embase, Cochrane Library and CNKI database, with the key words of "child" "adolescent" "obesity" "overweight" "pediatric obesity" "weight loss" "dietary protein" "dietary carbohydrate" "caloric restrict" both in English and Chinese. The quality of the included literature was evaluated according to the "risk of bias" assessment tool, which included bias from the randomization process, deviation from intended interventions, missing outcome data, measurement of the outcome and selection of the reported results. Moreover, calculated the pooled mean difference, perform heterogeneity test, and assess publication bias. Results: A total of 8 articles were selected from the retrieved 4 836 articles, all in English. The sample sizes ranged from 4 to 120. The analysis showed that the post-intervention body mass index (mean difference (MD)=-0.66, 95%CI -1.76-0.44), body mass index Z-scores (MD=-0.09, 95%CI-0.23-0.05), fat content percentage (MD=-1.07, 95%CI-2.88-0.74), high density lipoprotein (MD=0.02, 95%CI-0.02-0.06) and low density lipoprotein (MD=0.04, 95%CI-0.08-0.17) were not significantly different with those of the standard protein diet group, with P values being 0.240, 0.220, 0.250, 0.360 and 0.480, respectively. Subgroup analysis showed that after excluding one study, the difference in body mass index between the short-term intervention group and control group was statistically significant (MD=-1.60, 95%CI-3.14--0.06, P=0.040). Conclusions: A short-term high-protein diet intervention seems to improve the body mass index status of overweight and obese children. Nevertheless, a high-protein diet does not affect any other selected anthropometric indices and blood lipids. More studies with large sample sizes, higher quality and comparable standard of high-protein diet are needed for further demonstration.
Adolescent ; Child ; Diet, High-Protein ; Humans ; Lipids ; Overweight ; Pediatric Obesity ; Weight Loss

OBJECTIVE: To investigate the effect of Yinlai Decoction (YD) on the microstructure of colon, and activity of D-lactic acid (DLA) and diamine oxidase (DAO) in serum of pneumonia mice model fed with high-calorie and high-protein diet (HCD). METHODS: Sixty male Kunming mice were randomly divided into 6 groups by the random number table method: normal control, pneumonia, HCD, HCD with pneumonia (HCD-P), YD (229.2 mg/mL), and dexamethasone (15.63 mg/mL) groups, with 10 in each group. HCD mice were fed with 52% milk solution by gavage. Pneumonia mice was modeled with lipopolysaccharide inhalation and was fed by gavage with either the corresponding therapeutic drugs or saline water, twice daily, for 3 days. After hematoxylin-eosin staining, the changes in the colon structure were observed under light microscopy and transmission electron microscope, respectively. Enzyme-linked immunosorbent assay was used to detect the protein levels of DLA and DAO in the serum of mice. RESULTS: The colonic mucosal structure and ultrastructure of mice in the normal control group were clear and intact. The colonic mucosal goblet cells in the pneumonia group tended to increase, and the size of the microvilli varied. In the HCD-P group, the mucosal goblet cells showed a marked increase in size with increased secretory activity. Loose mucosal epithelial connections were also observed, as shown by widened intercellular gaps with short sparse microvilli. These pathological changes of intestinal mucosa were significantly reduced in mouse models with YD treatment, while there was no significant improvement after dexamethasone treatment. The serum DLA level was significantly higher in the pneumonia, HCD, and HCD-P groups as compared with the normal control group (P<0.05). Serum DLA was significantly lower in the YD group than HCD-P group (P<0.05). Moreover, serum DLA level significantly increased in the dexamethasone group as compared with the YD group (P<0.01). There was no statistical significance in the serum level of DAO among groups (P>0.05). CONCLUSIONS YD can protect function of intestinal mucosa by improving the tissue morphology of intestinal mucosa and maintaining integrity of cell connections and microvilli structure, thereby reducing permeability of intestinal mucosa to regulate the serum levels of DLA in mice.
Mice ; Male ; Animals ; Lactic Acid/pharmacology* ; Intestinal Mucosa ; Colon/pathology* ; Dexamethasone/pharmacology* ; Diet, High-Protein ; Pneumonia/pathology*

Obesity has been reported to be associated with low grade inflammatory status. In this study, we investigated the inflammatory response as well as associated signaling molecules in immune cells from diet-induced obese mice. Four-week-old C57BL mice were fed diets containing 5% fat (control) or 20% fat and 1% cholesterol (HFD) for 24 weeks. Splenocytes (1 x 10(7) cells) were stimulated with 10 microg/mL of lipopolysaccharide (LPS) for 6 or 24 hrs. Production of interleukin (IL)-1beta, IL-6, and TNF-alpha as well as protein expression levels of nucleotide-binding oligomerization domain (NOD)2, signal transducer and activator of transcription (STAT)3, and pSTAT3 were determined. Mice fed HFD gained significantly more body weight compared to mice fed control diet (28.2 +/- 0.6 g in HFD and 15.4 +/- 0.8 g in control). After stimulation with LPS for 6 hrs, production of IL-1beta was significantly higher (P = 0.001) and production of tumor necrosis factor (TNF)-alpha tended to be higher (P < 0.064) in the HFD group. After 24 hrs of LPS stimulation, splenocytes from the HFD group produced significantly higher levels of IL-6 (10.02 +/- 0.66 ng/mL in HFD and 7.33 +/- 0.56 ng/mL in control, P = 0.005) and IL-1beta (121.34 +/- 12.72 pg/mL in HFD and 49.74 +/- 6.58 pg/mL in control, P < 0.001). There were no significant differences in the expression levels of STAT3 and pSTAT3 between the HFD and the control groups. However, the expression level of NOD2 protein as determined by Western blot analysis was 60% higher in the HFD group compared with the control group. NOD2 contributes to the induction of inflammation by activation of nuclear factor kappaB. These findings suggest that diet-induced obesity is associated with increased inflammatory response of immune cells, and higher expression of NOD2 may contribute to these changes.
Animals ; Blotting, Western ; Body Weight ; Cholesterol ; Diet ; Diet, High-Fat ; Inflammation ; Interleukin-6 ; Interleukins ; Mice ; Mice, Inbred C57BL ; Mice, Obese ; Obesity ; Staphylococcal Protein A ; Transducers ; Tumor Necrosis Factor-alpha

The present study was to investigate the effects of Lycii Cortex Radicis (LCR), the root bark of lycium (Lycium chenese Miller) and ginger (Gin) on body lipid status and serum levels of cytokines. Sprague-Dawley (SD) male rats weighing 193.6 +/- 16.8 g were divided into five groups, including one low fat (LF) and four high fat groups, i.e. HF-Control, HF-LCR, HF-Gin and HF-LCR + Gin groups. Diets for HF-LCR, HF-Gin and HF-LCR + Gin groups contained purified extracts having 0.2 g LCR tyramine, ginerol and 0.1 g tyramine plus 0.02 g gingerol per kg, respectively. Compared with those of the HF-Control total serum cholesterol level decreased, and HDL-cholesterol level increased in the HF-LCR group and serum triglyceride levels decreased in the three experimental groups fed the purified extracts. Liver cholesterol level was lower in the HF-LCR group than the HF-Control group, but triglyceride levels, which were increased by high fat diets were not changed by significantly by LCR or ginger extracts. Fecal lipid excretion was higher in the HF-LCR and HF-Gin groups, but cholesterol excretion was lower in the HF-Gin group than in the HF-Control group. The activities of liver cytosolic glucose-6-phosphate dehydrogenase and malic enzyme were lower in the HF-LCR + Gin group than in the HF-Control group. Serum adiponectin levels did not differ among the five groups, while leptin level was lower in the HF-Gin group and C-reactive protein levels were lower in the HF-Gin and the HF-LCR + Gin groups than in the HF-Control group. It is concluded that LCR can be utilized as an ingredient for lipid-lowering functional foods in the form of purified extract and addition of small amount of ginger extract would be useful for reducing one of the inflammatory cytokines to help prevent atherosclerosis.
Adiponectin ; Animals ; Atherosclerosis ; C-Reactive Protein ; Catechols ; Cholesterol ; Cytokines ; Cytosol ; Diet ; Diet, High-Fat ; Fatty Alcohols ; Functional Food ; Ginger ; Glucosephosphate Dehydrogenase ; Humans ; Leptin ; Liver ; Lycium ; Male ; Rats ; Tyramine

Movement defects in obesity are associated with peripheral muscle defects, arthritis, and dysfunction of motor control by the brain. Although movement functionality is negatively correlated with obesity, the brain regions and downstream signaling pathways associated with movement defects in obesity are unclear. A dopaminergic neuronal pathway from the substantia nigra (SN) to the striatum is responsible for regulating grip strength and motor initiation through tyrosine hydroxylase (TH) activity-dependent dopamine release. We found that mice fed a high-fat diet exhibited decreased movement in open-field tests and an increase in missteps in a vertical grid test compared with normally fed mice. This motor abnormality was associated with a significant reduction of TH in the SN and striatum. We further found that phosphorylation of c-Jun N-terminal kinase (JNK), which modulates TH expression in the SN and striatum, was decreased under excess-energy conditions. Our findings suggest that high calorie intake impairs motor function through JNK-dependent dysregulation of TH in the SN and striatum.
Animals ; Arthritis ; Brain ; Diet, High-Fat* ; Dopamine ; Dopaminergic Neurons* ; Hand Strength ; JNK Mitogen-Activated Protein Kinases ; Mesencephalon* ; Mice ; Obesity ; Phosphorylation ; Substantia Nigra ; Tyrosine 3-Monooxygenase

Gut microbiota dysbiosis is an avenue for the promotion of atherosclerosis (AS) and this effect is mediated partly via the circulating microbial metabolites. More microbial metabolites related to AS vascular inflammation, and the mechanisms involved need to be clarified urgently. Paeonol (Pae) is an active compound isolated from Paeonia suffruticoas Andr. with anti-AS inflammation effect. However, considering the low oral bioavailability of Pae, it is worth exploring the mechanism by which Pae reduces the harmful metabolites of the gut microbiota to alleviate AS. In this study, ApoE^-/- mice were fed a high-fat diet (HFD) to establish an AS model. AS mice were administrated with Pae (200 or 400 mg·kg^-1) by oral gavage and fecal microbiota transplantation (FMT) was conducted. 16S rDNA sequencing was performed to investigate the composition of the gut microbiota, while metabolomics analysis was used to identify the metabolites in serum and cecal contents. The results indicated that Pae significantly improved AS by regulating gut microbiota composition and microbiota metabolic profile in AS mice. We also identified α-hydroxyisobutyric acid (HIBA) as a harmful microbial metabolite reduced by Pae. HIBA supplementation in drinking water promoted AS inflammation in AS mice. Furthermore, vascular endothelial cells (VECs) were cultured and stimulated by HIBA. We verified that HIBA stimulation increased intracellular ROS levels, thereby inducing VEC inflammation via the TXNIP/NLRP3 pathway. In sum, Pae reduces the production of the microbial metabolite HIBA, thus alleviating the ROS/TXNIP/NLRP3 pathway-mediated endothelial inflammation in AS. Our study innovatively confirms the mechanism by which Pae reduces the harmful metabolites of gut microbiota to alleviate AS and proposes HIBA as a potential biomarker for AS clinical judgment.
Animals ; Mice ; Atherosclerosis/drug therapy* ; Diet, High-Fat ; Endothelial Cells ; Inflammation/drug therapy* ; Mice, Inbred C57BL ; NLR Family, Pyrin Domain-Containing 3 Protein/genetics* ; Reactive Oxygen Species

We sought to identify common key regulators and build a gene-metabolite network in different nonalcoholic fatty liver disease (NAFLD) phenotypes. We used a high-fat diet (HFD), a methionine-choline-deficient diet (MCDD) and streptozocin (STZ) to establish nonalcoholic fatty liver (NAFL), nonalcoholic steatohepatitis (NASH) and NAFL+type 2 diabetes mellitus (T2DM) in rat models, respectively. Transcriptomics and metabolomics analyses were performed in rat livers and serum. A functional network-based regulation model was constructed using Cytoscape with information derived from transcriptomics and metabolomics. The results revealed that 96 genes, 17 liver metabolites and 4 serum metabolites consistently changed in different NAFLD phenotypes (>2-fold, P<0.05). Gene-metabolite network analysis identified ccl2 and jun as hubs with the largest connections to other genes, which were mainly involved in tumor necrosis factor, P53, nuclear factor-kappa B, chemokine, peroxisome proliferator activated receptor and Toll-like receptor signaling pathways. The specifically regulated genes and metabolites in different NAFLD phenotypes constructed their own networks, which were mainly involved in the lipid and fatty acid metabolism in HFD models, the inflammatory and immune response in MCDD models, and the AMPK signaling pathway and response to insulin in HFD+STZ models. Our study identified networks showing the general and specific characteristics in different NAFLD phenotypes, complementing the genetic and metabolic features in NAFLD with hepatic and extra-hepatic manifestations.
AMP-Activated Protein Kinases ; Animals ; Complement System Proteins ; Diabetes Mellitus ; Diet ; Diet, High-Fat ; Insulin ; Liver ; Metabolism ; Metabolomics ; Models, Animal ; Non-alcoholic Fatty Liver Disease* ; Peroxisomes ; Phenotype ; Rats ; Streptozocin ; Toll-Like Receptors ; Tumor Necrosis Factor-alpha

BACKGROUD/OBEJECTIVES: The mechanism of how black garlic effects lipid metabolism remains unsolved. Therefore, the objectives of this study were to determine the effects of black garlic on lipid profiles and the expression of related genes in rats fed a high fat diet. MATERIALS/METHODS: Thirty-two male Sqrague-Dawley rats aged 4 weeks were randomly divided into four groups (n=8) and fed the following diets for 5 weeks: normal food diet, (NF); a high-fat diet (HF); and a high-fat diet + 0.5% or 1.5% black garlic extract (HFBG0.5 or HFBG1.5). Body weights and blood biochemical parameters, including lipid profiles, and expressions of genes related to lipid metabolism were determined. RESULTS: Significant differences were observed in the final weights between the HFBG1.5 and HF groups. All blood biochemical parameters measured in the HFBG1.5 group showed significantly lower values than those in the HF group. Significant improvements of the plasama lipid profiles as well as fecal excretions of total lipids and triglyceride (TG) were also observed in the HFBG1.5 group, when compared to the HF diet group. There were significant differences in the levels of mRNA of sterol regulatory element binding protein-1c (SREBP-1c), acetyl-CoA carboxylase (ACC), fatty acid synthase (FAS), and glucose-6-phosphate dehydrogenase (G6PDH) in the HFBG1.5 group compared to the HF group. In addition, the hepatic expression of (HMG-CoA) reductase and Acyl-CoA cholesterol acyltransferase (ACAT) mRNA was also significantly lower than the HF group. CONCLUSIONS: Consumption of black garlic extract lowers SREBP-1C mRNA expression, which causes downregulation of lipid and cholestrol metahbolism. As a result, the blood levels of total lipids, TG, and cholesterol were decreased.
Acetyl-CoA Carboxylase ; Animals ; Body Weight ; Cholesterol ; Diet ; Diet, High-Fat* ; Down-Regulation ; Garlic* ; Glucosephosphate Dehydrogenase ; Humans ; Lipid Metabolism* ; Male ; Oxidoreductases ; Rats* ; RNA, Messenger ; Sterol O-Acyltransferase ; Sterol Regulatory Element Binding Protein 1 ; Triglycerides ; Weights and Measures

Aged garlic extract (AGE) is known to have a protective effect against immune system, endothelial function, oxidative stress and inflammation. We examined the effects of exercise with and without aged garlic extract administration on body weight, lipid profiles, inflammatory cytokines, and oxidative stress marker in high-fat diet (HFD)-induced obese rats. Forty-five Sprague-Dawley rats were fed either a HFD (HFD, n = 40) or a normal diet (ND, n = 5) for 6 weeks and thereafter randomized into ND (n = 5), HFD (n = 10), HFD with AGE (n = 10), HFD with Exercise (n = 10), or HFD with Exercise+AGE (n = 10) for 4 weeks. AGE groups were administered at a dose of 2.86 g/kg.body weight, orally. Exercise consisted of running 15-60 min 5 days/week with gradually increasing intensity. AGE (P < 0.01), Exercise, and Exercise+AGE (P < 0.001) attenuated body weight gain and food efficiency ratio compared to HFD. Visceral fat and liver weight gain were attenuated (P < 0.05) with all three interventions with a greater effect on visceral fat in the Exercise+AGE than AGE (P < 0.001). In reducing visceral fat (P < 0.001), epididymal fat (P < 0.01) and liver weight (P < 0.001), Exercise+AGE was effective, but exercise showed a stronger suppressive effect than AGE. Exercise+AGE showed further additive effects on reducing visceral fat and liver weight (P < 0.001). AGE significantly attenuated the increase in total cholesterol and low-density lipoprotein-cholesterol compared with HFD (P < 0.05). Exercise+AGE attenuated the increase in triglycerides compared with HFD (P < 0.05). Exercise group significantly decrease in C-reactive protein (P < 0.001). These results suggest that AGE supplementation and exercise alone have anti-obesity, cholesterol lowering, and anti-inflammatory effects, but the combined intervention is more effective in reducing weight gain and triglycerides levels than either intervention alone.
Aged ; Animals ; Body Weight ; C-Reactive Protein ; Cholesterol ; Cytokines ; Diet ; Diet, High-Fat ; Garlic ; Humans ; Immune System ; Inflammation ; Intra-Abdominal Fat ; Liver ; Obesity ; Oxidative Stress ; Rats ; Rats, Sprague-Dawley ; Running ; Triglycerides ; Weight Gain