Diet Feeding Research Articles

Functional study of amine oxidase copper-containing 1 (AOC1) in lipid metabolism

Amine oxidase copper-containing 1 (AOC1) is a key member of copper amine oxidase family, which is responsible for deamination oxidation of histamine and putrescine. In recent years, AOC1 has been reported to be associated with various cancers, with its expression levels significantly elevated in certain cancer cells, suggesting its potential role in cancer progression. However, its function in lipid metabolism still remains unclear. Through genetic analysis, we have discovered a potential relationship between AOC1 and lipid metabolism. To further investigate, we generated Aoc1 -/- mice and characterized their metabolic phenotypes on both chow diet and high-fat diet (HFD) feeding conditions. On HFD feeding conditions, Aoc1 -/- mice exhibited significantly higher fat mass and impaired glucose sensitivity, and lipid accumulation in white adipose tissue and liver was also increased. This study uncovers the potential role of AOC1 in lipid metabolism and its implications in metabolic disorders such as obesity and type 2 diabetes, providing new targets and research directions for treating metabolic diseases.

Evaluation of fenugreek (Trigonella foenum-graecum L.) powder supplementation on metabolic syndrome, oxidative stress and inflammation in high fat diet fed rats

As an anti-diabetic medicinal plant, fenugreek seed (FG) (Trigonella foenum-graecum L.) has long been utilized and has many therapeutic uses. The current investigation sought to ascertain the effects of FG powder supplementation on insulin resistance, dyslipidemia, and oxidative stress in rats given a high-fat (HF) diet. Measurements of biochemical and antioxidant indicators were made in animal blood and tissue samples, including malondialdehyde (MDA), nitric oxide (NO), advanced oxidation protein product (AOPP), reduced glutathione (GSH), superoxide dismutase (SOD), triglycerides (TG), and catalase (CAT). Furthermore, the tissues of the kidney, heart, and liver were stained histologically. HF diet feeding in rats declined in antioxidant enzyme activities, along with a rise in MDA and other indicators of oxidative stress, which were mitigated by the supplementation of FG. Additionally, FG supplementation enhanced the expression of genes corresponding to antioxidants in the liver of rats given HF diet. FG supplements in HF diet-fed rats displayed altered gene expression in the livers that metabolized fat. Histological staining of the liver demonstrated that FG supplementation reduced necrosis and the accumulation of fat droplets in the liver of rats given HF diets. In conclusion, this finding showed that FG supplementation in HF diet fed rats reduced the plasma lipids, decreased oxidative stress and raised antioxidant enzyme activities. FG may therefore aid in reducing the chronic complications linked to the HF diet in experimental rats.

Mechanisms of Action of Exclusive Enteral Nutrition and Other Nutritional Therapies in Crohn’s Disease

Background and Objectives: Crohn’s disease (CD) is an inflammatory bowel disease (IBD) characterized by transmural inflammation and intestinal fibrosis involving mostly the small intestine and colon. The pathogenic mechanisms of CD remain incompletely understood and cures are unavailable. Current medical therapies are aimed at inducing prolonged remission. Most of the medical therapies such as corticosteroids have substantial adverse effects. Consequently, many dietary therapies have been explored for the management of CD. Up to now, exclusive enteral nutrition (EEN) has been considered the only established dietary treatment for IBD, especially CD. In this article, we aim to give a concise review about the current therapeutic options and challenges in the management of CD and aim to compare the efficacy of EEN with other dietary therapies and update on the possible mechanisms of the benefits of EEN and other nutritional therapies. Methods: We searched the literature up to August 2024 through PubMed, Web of Science, and other sources using search terms such as EEN, nutritional therapy, IBD, Crohn’s disease, ulcerative colitis. Clinical studies in patients and preclinical studies in rodent models of IBD were included in the summary of the therapeutic benefits. Results and Conclusions: EEN involves oral or nasogastric tube feeding of a complete liquid diet with exclusion of normal foods for a defined period (usually 6 to 8 weeks). EEN treatment is demonstrated to have anti-inflammatory and healing effects in CD through various potential pathways, including altering gut bacteria and their metabolites, restoring the barrier function, direct anti-inflammatory action, and indirect anti-inflammatory action by eliminating mechanical stress in the bowel. However, efficacy of other nutritional therapies is not well established in CD, and mechanisms of action are largely unknown.

Targeting Unc5b in macrophages drives atherosclerosis regression and pro-resolving immune cell function

Atherosclerosis results from lipid-driven inflammation of the arterial wall that fails to resolve. Imbalances in macrophage accumulation and function, including diminished migratory capacity and defective efferocytosis, fuel maladaptive inflammation and plaque progression. The neuroimmune guidance cue netrin-1 has dichotomous roles in inflammation partly due to its multiple receptors; in atherosclerosis, netrin-1 promotes macrophage survival and retention via its receptor Unc5b. To minimize the pleiotropic effects of targeting netrin-1, we tested the therapeutic potential of deleting Unc5b in mice with advanced atherosclerosis. We generated Unc5bfl/flCx3cr1creERT2/WT mice, which allowed conditional deletion of Un5b (∆Unc5bMØ) in monocytes and macrophages by tamoxifen injection. After inducing advanced atherosclerosis by hepatic PCSK9 overexpression and western diet feeding for 20 wk, Unc5b was deleted and hypercholesterolemia was normalized to simulate clinical lipid management. Deletion of myeloid Unc5b led to a 40% decrease in atherosclerotic plaque burden and reduced plaque complexity compared to Unc5bfl/flCx3cr1WT/WT littermate controls (CtrlMØ). Consistently, plaque macrophage content was reduced by 50% in ∆Unc5bMØ mice due to reduced plaque Ly6Chi monocyte recruitment and macrophage retention. Compared to CtrlMØ mice, plaques in ∆Unc5bMØ mice had reduced necrotic area and fewer apoptotic cells, which correlated with improved efferocytotic capacity by Unc5b-deficient macrophages in vivo and in vitro. Beneficial changes in macrophage dynamics in the plaque upon Unc5b deletion were accompanied by an increase in atheroprotective T cell populations, including T-regulatory and Th2 cells. Our data identify Unc5b in advanced atherosclerosis as a therapeutic target to induce pro-resolving restructuring of the plaque immune cells and to promote atherosclerosis regression.

Renal denervation ameliorates atrial remodeling in type 2 diabetic rats by regulating mitochondrial dynamics.

There is no effective treatment for diabetes-related atrial remodeling currently. This study aimed to investigate the effects of renal denervation (RDN) on diabetes-related atrial remodeling and explore the related mechanisms. A type 2 diabetes mellitus model was established by high-fat diet feeding and low-dose streptozotocin injection in Sprague‒Dawley rats. After successful modeling, the diabetic rats were randomly assigned to two groups according to whether they were subjected to RDN or sham RDN surgery. At the end of the experiment, cardiac function and structure were evaluated by echocardiography and histology, respectively. Mitochondrial morphology, function and mitochondrial dynamics were assessed by multiple methods. Mdivi1 was used to verify the mechanism by which RDN improves atrial remodeling. In the 10th week, diabetic rats exhibited obvious atrial remodeling, including atrial enlargement and diastolic dysfunction. Pathological staining showed that diabetic rats had cardiomyocyte hypertrophy and interstitial fibrosis in atrial tissues. In terms of mitochondrial morphology and function, diabetic rats exhibited fragmented mitochondria, reduced adenosine triphosphate production and decreased mitochondrial membrane potential levels. Abnormal mitochondrial dynamics in diabetic rats were characterized by the inhibition of mitochondrial fusion, excessive mitochondrial fission, and the suppression of mitophagy. However, RDN effectively ameliorated diabetes-induced pathological atrial remodeling. In addition, RDN significantly improved mitochondrial morphological and functional abnormalities and corrected the disorders of mitochondrial dynamics. Furthermore, the protective effects of RDN against atrial remodeling were related to the regulation of mitochondrial dynamics. RDN prevented diabetes-induced atrial remodeling. These protective effects might be related to improvements in mitochondrial dynamics.

Calorie restriction increases insulin sensitivity to promote beta cell homeostasis and longevity in mice

Caloric restriction (CR) can extend the organism life- and health-span by improving glucose homeostasis. How CR affects the structure-function of pancreatic beta cells remains unknown. We used single nucleus transcriptomics to show that CR increases the expression of genes for beta cell identity, protein processing, and organelle homeostasis. Gene regulatory network analysis reveal that CR activates transcription factors important for beta cell identity and homeostasis, while imaging metabolomics demonstrates that beta cells upon CR are more energetically competent. In fact, high-resolution microscopy show that CR reduces beta cell mitophagy to increase mitochondria mass and the potential for ATP generation. However, CR beta cells have impaired adaptive proliferation in response to high fat diet feeding. Finally, we show that long-term CR delays the onset of beta cell aging hallmarks and promotes cell longevity by reducing beta cell turnover. Therefore, CR could be a feasible approach to preserve compromised beta cell structure-function during aging and diabetes.

Developing a More Sustainable Protein and Amino Acid Supply of Laying Hens in a Split Feeding System.

Two model experiments were conducted to investigate the different protein and amino acid supply of laying hens fed split feeding (SF) diets. In Experiment 1, one conventional (C) and one SF dietary treatment were established, and the diets were implemented for 12 weeks. The concentrations of crude protein, SID Lys, Met, Met + Cys Arg, Val, Thr, Leu, Ile, and Trp of the SF morning diet were the same as in the C diet. The crude protein content of the SF afternoon diet was lower (92%), while the SID values of Lys, Met, Met + Cys were identical compared to the C diet (100%). The SF treatment resulted in a reduced protein/N intake, better feed conversion ratio, higher eggshell thickness and apparent ileal digestibility of Asp, Leu, Lys, Gly, and Ser, and lower concentration of N forms (total, fecal, NH4+, uric acid, urinary) in the excreta compared to the C treatment. In Experiment 2, a control and a low protein (-2% crude protein but the same SID values of Lys, Met, Met + Cys, Thr, Val) SF treatment were compared for 6 weeks. The low protein SF treatment led to a decreased protein/N intake, higher laying rate, lower egg weight, higher ileal digestibility of Ala, Asp, Leu, and Ile, and similar N emission compared to the control SF treatment.

Medium-chain fatty acid receptor GPR84 deficiency leads to metabolic homeostasis dysfunction in mice fed high-fat diet.

Overconsumption of food, especially dietary fat, leads to metabolic disorders such as obesity and type 2 diabetes. Long-chain fatty acids, such as palmitoleate are recognized as the risk factors for these disorders owing to their high-energy content and lipotoxicity. In contrast, medium-chain fatty acids (MCFAs) metabolic benefits; however, their underlying molecular mechanisms remain unclear. GPR84 is an MCFA receptor, particularly for C10:0. Although evidence from invitro experiments and oral administration of C10:0 in mice suggests that GPR84 is related to the metabolic benefits of MCFAs via glucose metabolism, its precise roles invivo remain unclear. Therefore, the present study investigated whether GPR84 affects glucose metabolism and metabolic function using Gpr84-deficient mice. Although Gpr84-deficient mice were lean and had increased endogenous MCFAs under high-fat diet feeding conditions, they exhibited hyperglycemia and hyperlipidemia along with lower plasma insulin and glucagon-like peptide-1 (GLP-1) levels compared with wild-type mice. Medium-chain triglyceride (C10:0) intake suppressed obesity, and improved plasma glucose and lipid levels, and increased plasma GLP-1 levels in wild-type mice; however, these effects were partially attenuated in Gpr84-deficient mice. Our results indicate that long-term MCFA-mediated GPR84 activation improves the dysfunction of glucose and lipid homeostasis. Our findings may be instrumental for future studies on drug development with GPR84 as a potential target, thereby offering new avenues for the treatment of metabolic disorders like obesity and type 2 diabetes.

ADAM8 promotes alcoholic liver fibrosis through the MAPK signaling pathway

The effect and molecular regulatory mechanism of A Disintegrin and Metalloproteinase 8 (ADAM8) were explored in alcoholic liver fibrosis (ALF). C57BL/6N male mice were randomly divided into control, alcohol, and ADAM8-sgRNA3 plasmid groups. The control group received control liquid diet, while the alcohol and ADAM8-sgRNA3 plasmid groups were given alcohol liquid feed diet combined with ethanol gavage treatment for 8 weeks to induce ALF modeling. In addition, the ADAM8-sgRNA3 plasmid group was injected with the effective ADAM8-sgRNA3 plasmid, while the alcohol and control group mice were injected with an equivalent amount of physiological saline. LX-2 human hepatic stellate cells were divided into control, alcohol, si-ADAM8-2, and si-ADAM8-NC groups and induced for 48 h for model establishment in vitro. Serological detection, pathological staining, Western blotting, qRT-PCR and CCK8 assay were performed for experiments. Compared with the alcohol group, ADAM8 mRNA, protein and, positive area rate, serological indicators, pathological changes, and the expression of liver fibrosis marker and MAPK signaling pathway-related factors in the ADAM8-sgRNA3 plasmid group significantly decreased in vivo. Compared with the alcohol group, ADAM8 mRNA and protein expression, cell viability, and the expression of liver fibrosis markers and MAPK signaling pathway-related factors (p-ERK1/2, PCNA, Bcl-2, p-c-Jun, TGFβ1, p–p38 MAPK and HSP27) reduced significantly in the si-ADAM8-2 group. Therefore, ADAM8 promotes ALF through the MAPK signaling pathway, a promising target for treating ALF.

Non-mitogenic FGF19 mRNA-based therapy for the treatment of experimental metabolic dysfunction-associated steatotic liver disease (MASLD).

Metabolic dysfunction-associated steatohepatitis (MASH) represents a global health threat. MASH pathophysiology involves hepatic lipid accumulation and progression to severe conditions like cirrhosis and, eventually, hepatocellular carcinoma. Fibroblast growth factor (FGF)-19 has emerged as a key regulator of metabolism, offering potential therapeutic avenues for MASH and associated disorders. We evaluated the therapeutic potential of non-mitogenic (NM)-FGF19 mRNA formulated in liver-targeted lipid nanoparticles (NM-FGF19-mRNAs-LNPs) in C57BL/6NTac male mice with diet-induced obesity and MASH (DIO-MASH: 40% kcal fat, 20% kcal fructose, 2% cholesterol). After feeding this diet for 21 weeks, NM-FGF19-mRNAs-LNPs or control (C-mRNA-LNPs) were administered (0.5 mg/kg, i.v.) weekly for another six weeks, in which diet feeding continued. NM-FGF19-mRNAs-LNPs treatment in DIO-MASH mice resulted in reduced body weight, adipose tissue depots, and serum transaminases, along with improved insulin sensitivity. Histological analyses confirmed the reversal of MASH features, including steatosis reduction without worsening fibrosis. NM-FGF19-mRNAs-LNPs reduced total hepatic bile acids (BAs) and changed liver BA composition, markedly influencing cholesterol homeostasis and metabolic pathways as observed in transcriptomic analyses. Extrahepatic effects included the down-regulation of metabolic dysfunction-associated genes in adipose tissue. This study highlights the potential of NM-FGF19-mRNA-LNPs therapy for MASH, addressing both hepatic and systemic metabolic dysregulation. NM-FGF19-mRNA demonstrates efficacy in reducing liver steatosis, improving metabolic parameters, and modulating BA levels and composition. Given the central role played by BA in dietary fat absorption, this effect of NM-FGF19-mRNA may be mechanistically relevant. Our study underscores the high translational potential of mRNA-based therapies in addressing the multifaceted landscape of MASH and associated metabolic perturbations.

Increased dietary protein rather than fiber supports key metabolic and intestinal tissue functions in pigs, without increasing post-weaning diarrhea.

The post-weaning period in pigs is a critical window where nutritional interventions are implemented to prevent post-weaning diarrhea (PWD) and antibiotic use. One common strategy is feeding of low protein diets immediately following weaning. This intervention may reduce protein fermentation and pathogen proliferation, therefore decreasing the incidence of post-weaning diarrhea. These effects may also be mitigated by providing dietary fiber. However, studies examining the role of protein and fiber on gastrointestinal microbiota and metabolism are complicated by the presence of other substrates, including polyphenols and antinutritional factors in complex ingredients. In this study, semi-purified diets formulated to meet nutrient requirements were fed to 40 weaned pigs (n = 10/diet) to examine the effects of high protein (HP), high fiber (HF), or both (HFHP) compared to a control (CON) diet with industry standard crude protein and fiber content. Critical alterations in host metabolism and cecal transcriptome were identified in response to the CON diet. Diets with lower protein levels (CON and HF) induced alteration in transcripts from the serine synthesis pathways and integrated stress response in cecal tissue alongside systemic increases in metabolic pathways related to lysine degradation. High protein diets did not induce increases in gastrointestinal pathogen abundance. These results challenge the practice of feeding low protein diets post-weaning, by demonstrating a detrimental effect on intestinal cell function and muscle accretion. This suggests that with careful ingredient selection, increased dietary protein post-weaning could better pig health and growth compared to a standard diet.

Characterization of six clinical drugs and dietary intervention in the non-obese CDAA-HFD mouse model of MASH and progressive fibrosis.

The choline-deficient L-amino acid defined-high fat diet (CDAA-HFD) mouse model is widely used in preclinical metabolic dysfunction-associated steatohepatitis (MASH) research. To validate the CDAA-HFD mouse, we evaluated disease progression and responsiveness to dietary and pharmacological interventions with semaglutide, lanifibranor, elafibranor, obeticholic acid (OCA), firsocostat and resmetirom.Disease phenotyping was performed in C57BL/6J mice fed CDAA-HFD for 3-20 weeks and ranked using the MASLD Human Proximity Score (MHPS). Semaglutide, lanifibranor, elafibranor, OCA, firsocostat or resmetirom were profiled as treatment intervention for 8 weeks, starting after 6 weeks of CDAA-HFD feeding. Semaglutide and lanifibranor were further evaluated as early (preventive) therapy for 9 weeks, starting 3 weeks after CDAA-HFD diet feeding. Additionally, benefits of dietary intervention (chow reversal) for 8 weeks were characterized following 6 weeks of CDAA-HFD feeding. CDAA-HFD mice demonstrated a non-obese phenotype with fast onset and progression of MASH and fibrosis, high similarity to human MASH-fibrosis, and tumor development after 20 weeks of diet-induction. Semaglutide and lanifibranor partially reversed fibrosis when administered as prevention, but not as treatment intervention. Elafibranor was the only interventional drug to improve fibrosis. In comparison, chow-reversal resulted in complete steatosis regression with improved liver inflammation and fibrosis in CDAA-HFD mice. CDAA-HFD mice recapitulate histological hallmarks of advanced MASH with progressive severe fibrosis, however, in the absence of a clinical translational obese dysmetabolic phenotype. CDAA-HFD mice are suitable for profiling drug candidates directly targeting hepatic lipid metabolism, inflammation, and fibrosis. The timing of pharmacological intervention is critical for determining antifibrotic drug efficacy in the model.

Atractylodes Lancea and Its Constituent, Atractylodin, Ameliorates Metabolic Dysfunction-Associated Steatotic Liver Disease via AMPK Activation.

Metabolic dysfunction-associated steatotic liver disease (MASLD), which encompasses a spectrum of conditions ranging from simple steatosis to hepatocellular carcinoma, is a growing global health concern associated with insulin resistance. Since there are limited treatment options for MASLD, this study investigated the therapeutic potential of Atractylodes lancea, a traditional herbal remedy for digestive disorders in East Asia, and its principal component, atractylodin, in treating MASLD. Following 8 weeks of high-fat diet (HFD) feeding, mice received oral doses of 30, 60, or 120 mg/kg of Atractylodes lancea. In HFD-fed mice, Atractylodes lancea treatment reduced the body weight; serum triglyceride, total cholesterol, and alanine aminotransferase levels; and hepatic lipid content. Furthermore, Atractylodes lancea significantly ameliorated fasting serum glucose, fasting serum insulin, and homeostatic model assessment of insulin resistance levels in response to HFD. Additionally, a glucose tolerance test demonstrated improved glucose homeostasis. Treatment with 5 or 10 mg/kg atractylodin also resulted in anti-obesity, anti-steatosis, and glucose-lowering effects. Atractylodin treatment resulted in the downregulation of key lipogenic genes (Srebf1, Fasn, Scd2, and Dgat2) and the upregulation of genes regulated by peroxisome proliferator-activated receptor-α. Notably, the molecular docking model suggested a robust binding affinity between atractylodin and AMP-activated protein kinase (AMPK). Atractylodin activated AMPK, which contributed to SREBP1c regulation. In conclusion, our results revealed that Atractylodes lancea and atractylodin activated the AMPK signaling pathway, leading to improvements in HFD-induced obesity, fatty liver, and glucose intolerance. This study suggests that the phytochemical, atractylodin, can be a treatment option for MASLD.

Effects of dietary supplementation by modified palygorskite and essential oil/palygorskite complex on growth performance and intestinal flora composition of broilers with diarrhea

With the development trend of the industry, it can be seen that the substitution of antibiotics and reduction of zinc oxiden is still the hot spot of the industry. Diarrhea and inflammation occur frequently during livestock and poultry production, which is difficult to control. This experiment aimed to explore the effects and mechanisms of dietary supplementation of modified palygorskite (Mpal) and essential oil/ palygorskite composite (EO-PGS) on disease resistance and intestinal inflammatory damage in diarrhea broiler. In this experiment, there were a total of 420 broilers of 10-day-old selected and divided into 7 groups (n = 60), which were the nondiarrhea group fed with basal diet (normal control, NC), the diarrhea group fed with basal diet (diarrhea control, DC), and the rest were the diarrhea test group (diarrhea), supplemented with 1 kg/t, 2 kg/t and 4 kg/t of essential oils/ palygorskite complex (EO-PGS 1kg/T, EO-PGS 2kg/T, EO-PGS 4kg/T) in the basal diet, respectively, and 2 kg/t, 4 kg/t modified palygorskite group (Mpal 2kg/T, Mpal 4kg/T) in the basal diets, respectively. The experiment lasted for 8 d. The results showed that compared to normal broilers, the diarrhea index of diarrhea broilers remained around 2.0 with persistent mild diarrhea during the test period. The duodenal epithelial cells were damaged and shed, goblet cells increased, inflammatory cells infiltrated, diffuse congestion and hemorrhage in lamina propria, the serum lipopolysaccharides (LPS) content, and malondialdehyde (MDA) content increased significantly (P < 0.05). The serum superoxide dismutase (SOD) activity and immunoglobulin-M (IgM) levels significantly decreased, while serum immunoglobulin-G (IgG) and complement 3 (C3) levels significantly increased (P < 0.05). The expression of inflammatory cytokines interleukin-1β (IL-1β), interleukin-6 (IL-6), tumor necrosis factor-α (TNF-α), and nuclear factor κB (NF-κB) in duodenal epithelial cells was significantly upregulated on d 5 (P < 0.05). The abundance of Bacteroides in the duodenum of diarrhea broilers was significantly decreased, while the abundance of Proteobacteria was significantly increased (P < 0.05). Feeding diets supplemented with EO-PSG and 4 kg/t Mpal increased the average weight of diarrhea broilers (P < 0.05), reduced diarrhea index, improved immunity by increasing serum IgG, IgM, C3 and complement 4 (C4) levels (P < 0.05), enhanced the activity of serum antioxidant enzyme glutathione peroxidase (GSH-PX) and SOD activity, reduced serum MDA content, serum LPS levels, and decreased the expression of proinflammatory factors in the duodenal epithelial cell on d 5 (P < 0.05), alleviated duodenal epithelial cell injury, hemorrhage, inflammation infiltration and intestinal injury of diarrhea broilers from d 5 to d 8. Meanwhile, supplemented with EO-PSG and Mpal in diets regulated the intestinal microbiota, significantly increased the abundance of Bacteroidetes and decreased the abundance of Proteobacteria at the phylum level (P < 0.05). Microbial richness and diversity of microbiota were significantly increased by feeding the diet supplemented with 2 kg/t EO-PGS. In the beta diversity of the intestinal flora of the diets supplemented with 4 kg/t Mpal and 2 kg/t EO-PGS, the microbial community composition could be relatively easily distinguished with NC and DC groups. As a result of LEfSe analysis, the diets supplemented with 2 kg/t EO-PGS f_Clostridiaceae and g_Coprococcus were enriched in the caecum of diarrhea broilers, and the diets supplemented with 4 kg/t Mpal o_Bacteroidales, f_Rikenllaceae and g_Peptococcus were enriched in caecum of diarrhea broilers, between normal and diarrhea broilers (P<0.05). In conclusion, dietary supplementation with EO-PGS and Mpal could improve disease resistance and alleviate intestinal inflammatory damage in diarrhea broilers, but the effect of 2 kg/t Mpal was not significant. It was recommended that 2 kg/t EO-PGS or 4 kg/t Mpal be added to the broilers' diet according to the degree of diarrhea, and continuous feeding for more than 5 d.

12614 Induction Of Gpt2 During Metabolic Stress Is Associated With Reduced Incretin Responsiveness And Decreased β-cell Resilience To Metabolic Stress

Abstract Disclosure: S. Sen: None. A.V. Rozo: None. M. Haemmerle: None. J. Roman: None. A.V. Scota: None. J.A. Christine: None. E.M. Morrow: None. S.A. Tersey: None. C.B. Newgard: None. N.M. Doliba: None. D.A. Stoffers: None. Elevated blood glucose and lipids compromise pancreatic β-cell survival and function, but the underlying molecular mechanisms are ill-understood. We previously found that mitochondrial glutamate-pyruvate transaminase 2 (GPT2) is induced by ER and glucolipotoxic (GLT) stress. Here, we find that islets from type 2 diabetic donors exhibited markedly elevated levels of GPT2 compared to those of non-diabetic donors. GLT stress induced GPT2 in MIN6 cells, primary murine, and human non-diabetic islets. We generated β-cell specific Gpt2-deficient (Gpt2βKO) and Gpt2 floxed control (Gpt2f/f) mice to study the in vivo role of Gpt2 in β-cells. Intraperitoneal (IP) glucose tolerance (GTT) of Gpt2βKO mice was minimally affected on chow diet; therefore, we placed Gpt2f/f and Gpt2βKO mice on 45 kcal% high-fat and matched control diets. At 1 week, HFD-fed Gpt2βKO mice exhibited reduced β-cell death, measured by droplet digital PCR of circulating unmethylated preproinsulin DNA and by TUNEL analysis. Over 37 weeks, HFD-fed Gpt2βKO mice showed lower random blood glucose similar to Gpt2f/f mice under a control diet. While IPGTT was modestly improved in HFD-fed Gpt2βKO compared to HFD-fed Gpt2f/f mice, insulin secretion was not enhanced. Remarkably, oral glucose administration markedly improved glucose tolerance and enhanced insulin secretion of control- and HFD-fed Gpt2βKO mice compared to Gpt2f/f mice, while GLP-1 and GIP levels were unaltered, suggesting enhanced incretin action on the β-cell. Indeed, Gpt2βKO mice (under both control and high-fat diets) were more responsive to administration of GLP1R agonist Exendin-4 (Ex4) than Gpt2f/f mice. Isolated Gpt2βKO islets secreted more insulin when stimulated by Ex4 and GIP but not acetylcholine. Perifusion assay revealed markedly enhanced insulin secretion of Gpt2βKO islets in response to a stepwise exposure to increasing Ex4 doses, indicating enhanced incretin sensitivity. Metabolic flux analysis of Gpt2βKO and Gpt2f/f islets exposed to uniformly labeled glucose (U-13C glucose) under low (2.8mM), high (12mM) glucose, and high glucose with Ex4 conditions revealed elevated levels of key TCA cycle intermediates in Gpt2βKO islets under high glucose and high glucose+Ex4 conditions, indicating increased TCA cycle flux that likely enhances insulin secretion. Gpt2βKO mice had higher β cell mass than Gpt2f/f littermates under control diet and HFD feeding. Bulk islet RNA-seq from Gpt2f/f and Gpt2βKO mice exposed to GLT revealed upregulation of genes responsible for cell cycle and division pathways and downregulation of apoptosis and ER stress genes, consistent with enhanced survival of Gpt2βKO islets under stress conditions. In summary, β-cell GPT2 is increased during metabolic stress and T2D. Its deficiency induces a metabolic reprogramming that improves incretin sensitivity and protects pancreatic β-cells from metabolic stress. Presentation: 6/2/2024

Rifaximin alleviates MCD diet-induced NASH in mice by restoring the gut microbiota and intestinal barrier

AimsDue to the increasing global incidence rate of nonalcoholic steatohepatitis (NASH) combined with the lack of effective treatment methods for this disease, there is an urgent need to find new treatment strategies. The aim of this study was to investigate the efficacy of rifaximin in preventing and treating NASH and the related mechanism. Materials and methodsA NASH model was constructed by feeding male C57BL/6 mice a methionine-choline-deficient (MCD) diet for 4 weeks. Rifaximin was administered for 1 week before MCD diet feeding or during the last week of MCD diet feeding to investigate its preventive or therapeutic effects. Liver pathology, hepatic enzyme levels and metabolic indices were measured to evaluate the effects of rifaximin on NASH. Intestinal barrier integrity was measured via the Ussing chamber system and western blotting. 16S rDNA sequencing was conducted to investigate the fecal microbiota composition. Western blotting was performed to evaluate peroxisome proliferator activated receptor (PPAR)α and PPARγ protein levels. Key findingsRifaximin effectively alleviated MCD diet-induced NASH. The microbiota composition in MCD diet-fed mice was significantly altered, and intestinal barrier integrity was disrupted. Dysbiosis and intestinal barrier dysfunction were reversed by rifaximin. In addition, rifaximin modulated PPARα and PPARγ expression in the liver. SignificanceRifaximin effectively alleviated MCD diet-induced NASH by restoring the gut microbiota and reversing intestinal barrier dysfunction, suggesting that rifaximin treatment is a new approach for preventing and treating NASH.

Serum Biochemical Profile, Intestinal and Liver Histomorphometry of Captive Broad-Snouted Caiman (Caiman latirostris) Fed With a Diet Enriched With Soybean (Glycine max).

The impact of plant-based diets on crocodilians is unclear. Serum profiles and histomorphometry provide valuable insights into their nutritional and physiological status. This study aims to elucidate the impact of three levels of soybean meal substitution combined chicken by-product minced on the growth and health of broad-snouted caiman (Caiman latirostris). The research assesses the effects of diets supplemented with soybean meal on the blood biochemical profile, intestinal histomorphometry, and hepatic parameters of C. latirostris, providing essential information for understanding on the implications of dietary changes in this species. Forty-eight 6-month-old broad-snouted caimans were assigned to three dietary groups (0%, 25%, 40% soybean meal). Over a period of 90 days, data on growth, food consumption, serum biochemical analysis, intestinal and hepatic morphometry were recorded. The results showed that diets containing higher levels of soybean meal did not significantly affect growth, feed intake or serum profiles of total protein, albumin and cholesterol. However, changes in intestinal morphology were observed, with longer and wider villi in the animals feed with diets with soybean meal, indicating a gradual adaptation to new feeding diets. The presence of soybean meal reduced serum glucose and triglyceride profiles and hepatic lipid accumulation without affecting macronutrient digestion and absorption, considered beneficial for the caiman's health. This study provides valuable insights into the inclusion of soybean meal in the diet of Caiman latirostris and its effects on the intestines, liver, and physiology. It also highlights the importance of considering nutritional management as a key tool in improving the well-being and health of crocodilians in captivity.

Statement complementing the EFSA Scientific Opinion on application (EFSA-GMO-NL-2015-126) for authorisation of food and feed containing, consisting of and produced from genetically modified soybean MON 87705 × MON 87708 × MON 89788.

Following a request from the European Commission, the GMO Panel assessed additional information related to the application for authorisation of food and feed containing, consisting of and produced from genetically modified soybean MON × MON 87708 × MON 89788 (EFSA-GMO-NL-2015-126). The applicant conducted a 90-day feeding study on GM soybean MON 87705 and provided a proposal for post-market monitoring considering the altered fatty acid profile of GM soybean MON 87705 × MON 87708 × MON 89788, to fulfil the deficiencies identified by EFSA GMO Panel, addressing elements that remained inconclusive from a previous EFSA scientific opinion issued in 2020. The GMO Panel concludes that the 90-day feeding study on GM soybean MON 87705 is in line with the requirements of Regulation (EU) No 503/2013 and that no treatment-related adverse effects were observed in rats after feeding diets containing soybean MON 87705 meals at 30% or 15% for 90 days. The GMO Panel reiterates the recommendation for a PMM for food in accordance with Regulation (EC) No 1829/2003 and Regulation (EU) No 503/2013 and concludes that the proposal provided by the applicant is in line with the recommendations described for the PMM plan of soybean MON 87705 × MON 87708 × MON 89788 in the adopted scientific opinion. Taking into account the previous assessment and the new information, the GMO Panel concludes that soybean MON 87705 × MON 87708 × MON 89788, as assessed in the scientific opinion on application EFSA-GMO-NL-2015-126 and in the supplementary toxicity study, is as safe as its non-GM comparator and the non-GM reference varieties tested and does not represent a nutritional concern in humans and animals, within the scope of this application.

Environmental impacts of the filamentous fungi Paecilomyces variotii (PEKILO®) as a novel protein source in feeds for Atlantic salmon (Salmo salar)

The present study investigated environmental impacts associated with greenhouse gas (GHG) emission, economic Fish in Fish out ratio (eFIFO), waste outputs, and food-feed competition of feeding diets containing the filamentous fungus Paecilomyces variotii (PEKILO®) to Atlantic salmon (Salmo salar) pre-smolts. To achieve this, a feeding trial was conducted using four experimental diets. The control diet was based on fishmeal, soy protein concentrate, and wheat gluten meal. Diets 1, 2, and 3 were formulated so that P. variotii replaced 5, 10, and 20 % of the crude protein (CP) content of the control diet originating mainly from fishmeal, soy protein concentrate, and wheat gluten meal. Dietary inclusion of P. variotii affected simulated environmental indices such as GHG emission (P < 0.001); this index was P. variotii-dose-dependent, as illustrated by negative linear models (P < 0.001, adjusted R-squared = 0.96). Dietary inclusion of P. variotii was negatively correlated with eFIFO (P < 0.001, adjusted R-squared = 0.82). Dietary inclusion of P. variotii reduced food-competition feedstuff (FCF) in the diets (P < 0.001), as well as the amount of FCF to produce one kg of Atlantic salmon (P < 0.001). Relative to the control diet, solid nitrogen waste output, expressed as g kg−1 fish produced, was increased (P < 0.0001) in fish fed Diet 3. Conversely, linear decreases in solid phosphorous, magnesium and potassium (P < 0.0001) wastes expressed as g kg−1 fish produced were associated with increasing dietary inclusion of P. variotii. Likewise, linear decreases in dissolved nitrogen, magnesium, and potassium (P < 0.0001) wastes were observed with increasing replacement of the CP content of the control diet with P. variotii. Based on this assessment, P. variotii is a relevant alternative protein source with a low environmental footprint for use in Atlantic salmon feeds. Future novel feeds for Atlantic salmon should expand the use of alternative protein- and lipid-rich materials with a low ecological footprint that are produced according to a circular bioeconomy principle and utilize non-food-competing feedstuff (NFCF) such as P. variotii to move the industry toward sustainability.

Effect of management system and dietary seasonal variability on environmental efficiency and human net food supply of mountain dairy farming systems

Although mountain dairy cattle farming systems are pivotal for the economy, as well as for social and environmental aspects. They significantly contribute to the rural development which is currently strongly prioritized in the common EU agricultural policy, at the same time they are also increasingly criticized for having relatively high environmental impacts per kg of product, such as greenhouse gas emissions. Consequently, the aim of this study was to assess and compare the environmental efficiency of 2 common alpine dairy farming systems, with focus on the effects of grazing, considering the seasonal variability in feeding at individual cow level and farm management over a 3-year period. This study focuses on alpine farming systems but can be considered to represent well other topographically disadvantaged mountain areas. An intensively managed and globally dominating production system (high-input), aiming at high milk yield through relatively intensive feeding and the use of the high-yielding dual-purpose Simmental cattle, permanently confined in stables, was compared with a forage-based production system (low-input), based on seasonal grazing and the use of the autochthonous dual-purpose breed Tyrolean Grey. For the present analysis, a data set with information on feed intake and diet composition as well as animal productivity at individual cow level, and farm management data based on multiyear data recording was used. Four impact categories were quantified for 3 consecutive years: Global Warming Potential (GWP100), Acidification Potential (AP), Marine Eutrophication Potential (MEP), and Land Use (LU, m2yr and Pt, with the latter additionally considering the Soil Quality Index). Besides being attributed to 1 kg of fat and protein corrected milk (FPCM), these impact categories were also related to 1 m2 of on-farm area. Due to limited agronomic options beyond forage production and pasture use in alpine regions, net provision of protein was calculated for both farming systems to assess food supply and quantify the respective food-feed competition. Overall, the low-input farming system had greater environmental efficiency in terms of MEP per kg FPCM, as well as MEP and AP per m2 than the high-input system. LU was found to be consistently higher for the high-input than for the low-input system, the GWP100 per kg of FPCM was lower for the high-input system. Additionally, pasture access had a significant effect on the reduction of environmental impacts. Lastly, the net protein provision was slightly negative for the high-input system and marginally positive for the low-input system, indicating a lower food-feed competition for the latter. Future studies should also address the social and economic aspects of the farming systems, to offer a comprehensive overview of the 3 key factors necessary for achieving more sustainable farming systems, particularly in disadvantaged marginal regions such as mountain areas.