Comparison of the Protective Effects of Casein Hydrolysate Containing Tyr-Pro-Val-Glu-Pro-Phe and Casein on the Behaviors and Peripheral and Brain Functions in Mice with Chronic-Stress-Induced Anxiety and Insomnia.

Chronic stress is a major inducer of anxiety and insomnia. Milk casein has been studied for its stress-relieving effects. We previously prepared a casein hydrolysate (CP) rich in the sleep-enhancing peptide YPVEPF, and this study aims to systemically investigate the different protective effects of CP and casein on dysfunction and anxiety/insomnia behavior and its underlying mechanisms in chronically stressed mice. Behavioral results showed that CP ameliorated stress-induced insomnia and anxiety more effectively than milk casein, and this difference in amelioration was highly correlated with an increase in GABA, 5-HT, GABAA, 5-HT1A receptors, and BDNF and a decrease in IL-6 and NMDA receptors in stressed mice. Furthermore, CP restored these dysfunctions in the brain and colon by activating the HPA response, modulating the ERK/CREB-BDNF-TrκB signaling pathway, and alleviating inflammation. The abundant YPVEPF (1.20 ± 0.04%) and Tyr-based/Trp-containing peptides of CP may be the key reasons for its different effects compared to casein. Thus, this work revealed the main active structures of CP and provided a novel dietary intervention strategy for the prevention and treatment of chronic-stress-induced dysfunction and anxiety/insomnia behaviors.

Chlorate and perchlorate in tea leaves from major producing regions in China and related human exposure risk.

Chlorate and perchlorate are emerging pollutants that may interfere with thyroid function. Since they are highly water soluble, chlorate and perchlorate in tea leaves cause health concerns but have scarcely been studied. In this study, chlorate and perchlorate concentrations in 216 tea samples from different regions of China were determined. Perchlorate was detected in all the samples with a median concentration of 44.1 µg kg-1, while the chlorate detection frequency was 15.7%. We observed regional differences in perchlorate contents in tea leaves, with the highest quantity found in the central region of China. Except for dark tea, the concentration of perchlorate in tea infusions decreased with the increased number of times the tea leaves were brewed. The hazard quotients (HQs) of chlorate and perchlorate in all the samples were less than 1, suggesting negligible health risks caused by these pollutants from tea consumption. To the best of our knowledge, this is the first study to investigate chlorate and perchlorate contamination in tea infusions by simulating brewing behavior.

Anti-diabetic mechanism and potential bioactive peptides of casein hydrolysates in STZ/HFD-induced diabetic rats.

BACKGROUND: Casein hydrolysates have attracted much interest as anti-diabetic food, but their hypoglycemic mechanism and biopeptides are not well understood. This study aimed to explore the anti-diabetic mechanism and potential biopeptides of casein hydrolysates in streptozotocin/high-fat-diet-induced diabetic rats and HepG2 cells. RESULTS: Oral administration of casein hydrolysate prepared with papain-Flavourzyme combination (P-FCH) decreased fasting blood glucose, improved oral glucose tolerance, and reduced HbA1c values in diabetic rats. P-FCH was ineffective in alleviating insulin resistance (homeostasis model assessment and insulin sensitivity index) and enhancing hepatic insulin signaling transduction (phosphorylated Akt, hexokinase activity, and pyruvate kinase activity) in diabetic rats. However, P-FCH significantly upregulated adenosine monophosphate-activated protein kinase phosphorylation and glucose transporter-2 expression, inhibited phosphoenolpyruvate carboxylase kinase activity, and elevated glycogen content in liver tissue of diabetic rats. Furthermore, P-FCH increased glucose consumption independently in normal and insulin-resistant HepG2 cells without the presence of insulin. The peptide composition of P-FCH was characterized. The potential biopeptides in P-FCH showed the sequence characteristic of a Val at the N-terminal or a Pro at the P2 position, and the hypoglycemic activity of Val-Pro-Leu-Gly (the most potential biopeptide in P-FCH) was verified by oral glucose tolerance test in mice. CONCLUSION: These results suggested that activation of the non-insulin-mediated AMPK pathway might be the determinant mechanism of P-FCH on the hypoglycemic effect. The novel peptide Val-Pro-Leu-Gly in P-FCH was effective in reducing blood glucose levels when orally administered to mice. © 2023 Society of Chemical Industry.

Fucus vesiculosus polysaccharide alleviates type 2 diabetes in rats via remodeling gut microbiota and regulating glycolipid metabolism-related gene expression.

The antidiabetic activity and underlying mechanisms of Fucus vesiculosus polysaccharide (FVP) were studied in type 2 diabetic rats. Our results exhibited that FVP intervention reversed body weight loss, alleviated hyperglycemia and insulin resistance in diabetic rats. FVP also had the potential to ameliorate dyslipidemia, liver and kidney dysfunction, decrease oxidative stress, promote glycogen synthesis, and boost short-chain fatty acid production and total bile acid excretion. 16S rRNA gene sequencing analysis suggested that FVP interfered with the gut microbiota in a beneficial manner. Moreover, RT-qPCR results demonstrated that the antidiabetic activity of FVP in connection with the acceleration of blood glucose absorption and glycogen synthesis, the inhibition of gluconeogenesis, and the regulation of lipid metabolism in the liver. These findings suggested that FVP had antidiabetic effects on high-fat diet and STZ-induced diabetic rats and could be a potential resource for treating type 2 diabetes mellitus.

In Vitro Simulated Gastrointestinal Digestion Stability of a Neuroprotective Octapeptide WCPFSRSF and Prediction of Potential Bioactive Peptides in Its Digestive Fragments by Multiple Bioinformatics Tools.

WCPFSRSF, an octapeptide (Trp-Cys-Pro-Phe-Ser-Arg-Ser-Phe), has been reported to improve memory in mice, but its gastrointestinal stability is unclear. The objective of this study was to evaluate the gastrointestinal stability of peptide WCPFSRSF and explore the neuroprotective potential of its digestive fragments. Results showed that the content of WCPFSRSF after gastric and gastrointestinal digestion decreased to 71.64% and less than 1%, respectively. Furthermore, the antioxidant and neuroprotective ability of WCPFSRSF were also affected. Eleven and nine peptides were identified in its gastric and gastrointestinal digestive products, respectively. Multiple bioinformatics tools in combination with principal component analysis were employed to assess the physicochemical and structural properties of peptides. Novel peptides generated after gastrointestinal digestion could be classified into three groups: the first group had high bioactivity and bioavailability; the second group had high amphiphilicity, charge, and net hydrogen; and the third group had a long peptide chain. In addition, the representative peptides WCPF and SR showed neuroprotective ability.

1H NMR-based urinary metabolic analysis of high-dose cyclophosphamide-induced toxicity in mice.

Cyclophosphamide (CP) is widely used in clinical fields. Beside its therapeutic effects, CP shows toxicity depending on dose and administration schedule. In this study, the urinary metabolic profiles were investigated in mice intraperitoneally injected with high-dose CP (150 mg/kg body weight) once a week over four weeks using nuclear magnetic resonance (NMR)-based metabolomics. Twenty-six metabolites were identified as potential biomarkers by multivariate statistical analysis. A decrease in isoleucine, alanine, N-acetylglutamic acid, proline, methionine, valine, phenylacetylglulamine, dimethylamine, hippurate, acetic acid, lactate, α-oxoglutarate, citrate, malonic acid, creatinine, niacin, ß-hydroxybutyrate, and betaine, whereas an increase in leucine, glutamate, glycine, taurine, phenylacetylglycine, glucose, creatine, and choline were observed in the urine of high-dose CP-treated mice. Metabolites related to amino acid metabolism, energy metabolism, and gut microbial metabolism were changed markedly in the urine. Further metabolic pathway analysis suggested that seven metabolic pathways, including alanine, aspartate, and glutamate metabolism, arginine biosynthesis, glyoxylate, and dicarboxylate metabolism, glycine, serine and threonine metabolism, d-glutamine and d-glutamate metabolism, arginine, and proline metabolism, citrate cycle, as well as the gut microbiota metabolism, were significantly involved in response to high-dose CP treatment. These findings help to predict the toxicity of CP and understand the biological mechanism of the toxicity of CP.

Characterizations of food-derived ellagic acid-Undaria pinnatifida polysaccharides solid dispersion and its benefits on solubility, dispersity and biotransformation of ellagic acid.

The current study was aimed to enhance the solubility, dispersibility and biotransformation efficacy of ellagic acid (EA) by preparing food-derived ellagic acid-Undaria pinnatifida polysaccharides solid dispersion (EA/UPP SD). The results demonstrated that the solubility of EA/UPP SD was improved from 0.014 mg/mL to 0.383 mg/mL, and the enhancement was related to converting to a more amorphous state and restraining its self-aggregation during the mechanochemical process. The structure of EA/UPP SDs was mostly maintained by hydrogen bonds and hydrophobic interactions between EA and UPP. Moreover, the result of in vitro anaerobic incubations showed the biotransformation process was improved with EA/UPP SD addition to substrate due to the advance of microbial accessibility in EA dispersion. Altogether, these results indicated that the EA/UPP SDs expanded the application of EA by increasing the solubility and dispersity, and provided a theoretical basis for bioconversion efficiency enhancement.

Effects of glycation with chitooligosaccharide on digestion and fermentation processes of lactoferrin in vitro.

This study aimed to investigate the digestion and fermentation processes of lactoferrin (LF) glycated with chitooligosaccharide (COS) under a controlled Maillard reaction, utilizing the in vitro digestion and fermentation model, and to compare the results of these processes to LF undertaken without glycation. After gastrointestinal digestion, the products of the LF-COS conjugate were found to have more fragments with lower molecular weight than LF, and the antioxidant capabilities (via ABTS and ORAC assay) of the LF-COS conjugate digesta also increased. In addition, the undigested fractions could be further fermented by the intestinal microbiota. Compared with LF, more short-chain fatty acids (SCFAs) were generated (from 2397.40 to 2623.10 µg/g), and more species of microbiota (from 451.78 to 568.10) were observed in LF-COS conjugate treatment. Furthermore, the relative abundance of Bacteroides and Faecalibacterium that could utilize carbohydrates and metabolic intermediates to produce SCFAs also increased in LF-COS conjugate than that of LF. Our results demonstrated that glycation with COS under the controlled wet-heat treatment Maillard reaction could modify the digestion of LF and have a potentially positive influence on the intestinal microbiota community.

Dietary Supplement of Amomum villosum Lour. Polysaccharide Attenuates Ulcerative Colitis in BALB/c Mice.

Amomum villosum Lour. (A. villosum), a comestible medicinal plant, has been traditionally used in China to treat diarrhea, stomach fullness, and abdominal distension. Polysaccharide, the main chemical component of A. villosum, has been shown to possess potential antioxidant and glycosidase inhibitory activities; however, whether it has anticolitis activity is unknown. The aim of this research was to evaluate the anticolitis effects of A. villosum polysaccharide (AVLP) in BALB/c mice. The results showed that AVLP administration significantly reversed body weight loss, colon shortening and colon weight gain and decreased the levels of proinflammatory cytokines and chemokines in colitis mice (p < 0.05). AVLP administration also maintained intestinal barrier function by the upregulation of ZO-1 protein expression (p < 0.05). In addition, high-throughput sequencing analysis showed that AVLP possessed a great regulatory effect on the growth of Adlercreutzia, Clostridium, Streptococcus, Parabacteroides, Helicobacter, Odoribacter, and Alistipes (p < 0.05, LDA score > 2). The correlation analysis revealed that the protective effects against colitis of AVLP were highly correlated with intestinal bacterium regulation. These results suggest that AVLP intake could serve as a prospective nutritional strategy for inflammatory bowel diseases.

Essential oils of Zingiber officinale: Chemical composition, in vivo alleviation effects on TPA induced ear swelling in mice and in vitro bioactivities.

Zingiber officinale (ZO) is a traditional food condiment. The essential oils of Z. officinale (ZOEOs) are known to have multiple bioactivities. In this study, gas chromatography mass spectrometer (GC-MS) analytical method was used to identify active ingredient present in ZOEOs. A total of 41 compounds were identified in ZOEOs. Major components in ZOEOs were zingiberene (19.71%), (+)-ß-cedrene (12.85%), farnesene (12.17%), α-curcumene (10.18%) and ß-elemene (3.54%). Experimental results of 12-O-tetradecanoylphorbol-13 acetate (TPA) induced ear swelling validation mice model showed that ZOEOs treatment has better anti-inflammatory effect compared with ibuprofen (positive control) at high concentrations. Histological and immunohistochemical analysis showed that ZOEOs significantly decreased COX-2, IL-6 and NF-κB expression in a dose dependent manner. The mRNA levels of COX-2 and NF-κB were also down regulated by the application of ZOEOs. This indicated that ZOEOs exhibited positive effects in ear skin protection. Antibacterial experimental results showed that EOZOs had anti-bacterial effects on Escherichia coli, Pseudomonas aeruginosa, and Staphylococcus aureus. DPPH radical scavenging, A549 cell line and LNCaP cell line inhibition results indicated that ZOEOs exhibited potential antioxidant and anti-tumor properties. The findings of these study provide scientific basis on therapeutic use of ZO in food, cosmetic and pharmaceutical industries.

Simultaneous determination of twelve mycotoxins in edible oil, soy sauce and bean sauce by PRiME HLB solid phase extraction combined with HPLC-Orbitrap HRMS.

A solid phase extraction-high-performance liquid chromatography-tandem Orbitrap high resolution mass spectrometry (HPLC-Orbitrap HRMS) method was established for the determination of 12 mycotoxins (ochratoxin A, ochratoxin B, aflatoxin B1, aflatoxin B2, aflatoxin G1, aflatoxin G2, HT-2 toxin, sterigmatocystin, diacetoxysciroenol, penicillic acid, mycophenolic acid, and citreoviridin) in edible oil, soy sauce, and bean sauce. Samples were extracted by 80:20 (v:v) acetonitrile-water solution, purified by PRiME HLB column, separated by aQ C18 column with mobile phase consisting of 0.5 mmol/L ammonium acetate-0.1% formic acid aqueous solution and methanol. The results showed that the limits of detection (LODs) and limits of quantification (LOQs) of 12 mycotoxins were 0.12-1.2 µg/L and 0.40-4.0 µg/L, respectively. The determination coefficients of 12 mycotoxins in the range of 0.20-100 µg/L were > 0.998. The average recoveries in soy sauce and bean sauce were 78.4-106.8%, and the relative standard deviations (RSDs) were 1.2-9.7% under three levels, including LOQ, 2× LOQ and 10 × LOQ. The average recoveries in edible oil were 78.3-115.6%, and the precision RSD (n = 6) was 0.9-8.6%. A total of 24 edible oils, soy sauce and bean sauce samples were analyzed by this method. AFB1, AFB2, sterigmatocystin and mycophenolic acid were detected in several samples at concentrations ranging from 1.0 to 22.1 µg/kg. The method is simple, sensitive, and rapid and can be used for screening and quantitative analysis of mycotoxin contamination in edible oil, soy sauce, and bean sauce.

Effect of Bergamot and Laoxianghuang Polysaccharides on Gut Microbiota Derived from Patients with Hyperlipidemia: An Integrative Analysis of Microbiome and Metabolome during In Vitro Fermentation.

The aim of this study was to investigate the effects of bergamot polysaccharide (BP) and Laoxianghuang polysaccharides (LPs, fermented bergamot) on the microbiome and metabolome during the in vitro fermentation of gut microbiota from patients with hyperlipidemia. Results indicated that both BP and LPs were able to increase the production of acetic acid, propionic acid, and butyric acid. However, only LPs could decrease the content of isobutyric acid and isovaleric acid, which are detrimental to gut health. A 16S rRNA analysis showed that both BP and LPs could reduce the proportion of Fusobacterium, whereas they increased the Bacteroides content in hyperlipidemia. Untargeted UPLC-MS/MS metabolomic profiling found six bio-markers that were significantly changed after BP and LPs intervention, and four of the down-regulated metabolites were long-chain fatty acids associated with vascular diseases. These findings provide new evidence that BP and LPs have the potential to regulate imbalances in the gut microbiota in patients with hyperlipidemia and ameliorate its metabolic abnormalities.

The potential mechanisms of Macrocystis pyrifera polysaccharides mitigating type 2 diabetes in rats.

Our previous studies have proved that the anti-digestive polysaccharide from Macrocystis pyrifera possesses potential hypoglycemic and lipid-lowering activities; however, its potential mechanisms for improving diabetes have not been elucidated. The current study was aimed to determine the anti-diabetic effects and possible mechanisms of Macrocystis pyrifera polysaccharides (MPP) in diabetic rats. After 8-week MPP treatment, the serum profiles, gut bacteria composition and relative gene expressions of rats were determined. MPP administration effectively ameliorated the diabetic symptoms, dyslipidemia, liver and kidney damage, oxidative stress and chronic inflammation in diabetic rats. In addition, MPP treatment could also notably improve the microbial dysbiosis by increasing the beneficial bacteria and decreasing a bacterial pathogen in the diabetic rats. The RT-qPCR analysis indicated that MPP intervention significantly up-regulated the IRS/PI3K/AKT signaling pathway and down-regulated the relative expressions of glucose-6-phosphatase (G-6-Pase), phosphoenolpyruvate carboxykinase (PEPCK), acetyl-CoA carboxylase (ACC), hydroxymethylglutaryl CoA reductase (HMGCR) and sterol regulatory element binding protein 1c (SREBP-1c) in diabetic rats. These results demonstrated that MPP had the potential to be exploited as functional foods or pharmaceutical supplements for preventing and treating diabetes.

In vitro and in silico analysis of potential antioxidant peptides obtained from chicken hydrolysate produced using Alcalase.

Chicken hydrolysates (CHs) have been reported to protect mice against alcoholic liver injury possibly through oxidative stress reduction. In this study, the antioxidant activity of CHs was studied. Results showed that CHs exhibited significant antioxidant activity (around 600 and 400 µM TEAC/g in ORAC and ABTS assay, respectively) and could resist simulated gastrointestinal digestion. A total of 22 peptides were identified after antioxidant activity-oriented isolation using size-exclusion chromatography and high-performance liquid chromatography. Further in silico analysis and the validation of antioxidant activity revealed that novel peptides (RWGG and YYCQ) exhibited strong antioxidant activity. The most active peptide YYCQ displayed a TEAC value of 3.54 and 4.28 µM TEAC/µM in ORAC and ABTS assay, respectively. These peptides could contribute to reduce oxidative stress and protect against alcohol-induced liver injury. However, further studies understanding the bioactivity of such peptides in vivo are necessary before further applying them as functional food ingredient.

The Beneficial Effects of Two Polysaccharide Fractions from Sargassum fusiform against Diabetes Mellitus Accompanied by Dyslipidemia in Rats and Their Underlying Mechanisms.

The current study aimed to assess the anti-diabetic effects and potential mechanisms of two Sargassum fusiform polysaccharide fractions (SFPs, named SFP-1 and SFP-2). The carbohydrate-loading experiment revealed that SFP-2 could control postprandial hyperglycemia by inhibiting the activity of digestive enzymes in rats. The analysis of diabetic symptoms and serum profiles indicated that SFPs could mitigate diabetes accompanied by dyslipidemia, and SFP-2 showed better regulatory effects on body weight, food intake and the levels of total cholesterol (TC), triglycerides (TG), low density lipoprotein-cholesterol (LDL-C) and free fatty acid (FFA) in diabetic rats. Intestinal bacterial analysis showed that SFP treatment could reshape the gut flora of diabetic rats, and SFP-2 possessed a greater regulatory effect on the growth of Lactobacillus and Blautia than SFP-1. RT-qPCR analysis revealed that SFPs could regulate the genes involved in the absorption and utilization of blood glucose, hepatic glucose production and lipid metabolism, and the effects of SFP-2 on the relative expressions of Protein kinase B (Akt), Glucose-6-phosphatase (G-6-Pase), Glucose transporter 2 (GLUT2), AMP-activated protein kinase-α (AMPKα), Peroxisome proliferator-activated receptor γ (PPARγ) and Cholesterol 7-alpha hydroxylase (CYP7A1) were greater than SFP-1. All above results indicated that SFPs could be exploited as functional foods or pharmaceutical supplements for the treatment of diabetes and its complications.

Sargassum fusiforme polysaccharide is a potential auxiliary substance for metformin in the management of diabetes.

The present study investigated the positive effects of relatively low-dose metformin combined with Sargassum fusiforme polysaccharide (LMET-SFP) in high-fat diet and streptozotocin-induced diabetic rats, and explored the underlying mechanisms of LMET-SFP as compared to metformin alone in managing diabetes. The results indicate that both metformin and LMET-SFP can attenuate body weight loss and ameliorate hyperglycemia, insulin resistance and hyperlipidemia, and LMET-SFP exhibited better effects in lowering fasting blood glucose levels, insulin resistance index and serum cholesterol compared to metformin only. The administration of LMET-SFP could ameliorate liver dysfunction in diabetic rats. In addition, fecal bile acid data implied that LMET-SFP intervention contributed to an increase in fecal total bile acids, ursodesoxycholic acid and tauroursodesoxycholic acid profiles when compared to metformin treatment. Additionally, intestinal microbiological analysis showed that the acknowledged probiotics Lactobacillus and Bifidobacterium exhibited higher levels in the LMET-SFP group compared to the metformin group. RT-qPCR results demonstrated that the better hypoglycemic effects of LMET-SFP were mainly attributed to the down-regulation of 3-hydroxy-3-methylglutaryl-coenzyme A, cytosolic phosphoenolpyruvate carboxykinase and glucose-6-phosphatase expression, and the up-regulation of cholesterol 7α-hydroxylase expression, in contrast to metformin alone. These results suggest that SFP may be used as an auxiliary hypoglycemic substance for metformin in the future.

Mushroom polysaccharides with potential in anti-diabetes: Biological mechanisms, extraction, and future perspectives: A review.

Diabetes mellitus (DM) is a global health threat. Searching for anti-diabetic components from natural resources is of intense interest to scientists. Mushroom polysaccharides have received growing attention in anti-diabetes fields due to their advantages in broad resources, structure diversity, and multiple bioactivities, which are considered an unlimited source of healthy active components potentially applied in functional foods and nutraceuticals. In this review, the current knowledge about the roles of oxidative stress in the pathogenesis of DM, the extraction method of mushroom polysaccharides, and their potential biological mechanisms associated with anti-diabetes, including antioxidant, hypolipidemic, anti-inflammatory, and gut microbiota modulatory actions, were summarized based on a variety of in vitro and in vivo studies, with aiming at better understanding the roles of mushroom polysaccharides in the prevention and management of DM and its complications. Finally, future perspectives including bridging the gap between the intervention of mushroom polysaccharides and the modulation of insulin signaling pathway, revealing structure-bioactivity of mushroom polysaccharides, developing synergistic foods, conducting well-controlled clinical trials that may be very helpful in discovering valuable mushroom polysaccharides and better applications of mushroom polysaccharides in diabetic control were proposed.

Comparative study on the structural characterization and α-glucosidase inhibitory activity of polysaccharide fractions extracted from Sargassum fusiforme at different pH conditions.

Sargassum fusiforme polysaccharides (SFPs), including SFP-3-40, SFP-3-60, SFP-3-80, SFP-7-40, SFP-7-60, SFP-7-80, SFP-10-40, SFP-10-60, and SFP-10-80, were extracted at different pH (3, 7, and 10), and then precipitated with graded precipitation of 40%, 60% and 80% (v/v) ethanol solution, respectively. Their physicochemical properties and α-glucosidase inhibitory activity were determined. Results showed that SFPs significantly differed in the contents of total sugar, protein, uronic acid, sulfate, the zeta potential, and molecular weight distribution. SFPs, including SFP-10-40, SFP-10-60, and SFP-10-80, had bigger absolute zeta potential value and higher respective average molecular weight in the same ethanol concentration precipitate. All samples were mainly composed of fucose, glucuronic acid, and mannose with different molar ratios. The extraction pH and precipitation ethanol solution concentration caused little changes in functional groups, but significantly altered surface morphology of SFPs. Congo red test revealed that all polysaccharides were not helical polysaccharides. Rheological measurements indicated that SFPs were pseudoplastic fluids and showed elastic behavior of the gel. Except SFP-3-40 and SFP-3-60, all other samples had a stronger α-glucosidase inhibitory activity than that of acarbose. The inhibition type of SFPs against α-glucosidase varied owing to different extraction pH and precipitation ethyl concentration. This study shows that extraction pH can significantly affect the structure and hypoglycemic activity of SFPs and provide a data support for the scientific use of Sargassum fusiforme in industrial production.

The positive effects and underlying mechanisms of Undaria pinnatifida polysaccharides on type 2 diabetes mellitus in rats.

The aim of the current work was to investigate the anti-diabetic effects and underlying mechanisms of Undaria pinnatifida polysaccharides (UPP) based on a type 2 diabetes (T2DM) rat model. The starch loading test showed that UPP administration could reduce blood glucose fluctuations caused by eating. Analysis of diabetic symptoms and biochemical profiles showed that UPP intervention markedly decreased fasting blood glucose level, mitigated insulin resistance, improved glucose tolerance, dyslipidemia and liver and kidney damage in diabetic rats. The 16S rRNA analysis demonstrated that UPP intervention could markedly change the intestinal microflora composition, causing increases in Alistipes, Bacteroides, Christensenellaceae_R-7_group, Desulfovibrio, Muribaculaceae_norank, Ruminococcaceae_UCG-013, and Ruminococcaceae_UCG-014, and a decrease in Escherichia-Shigella. Furthermore, RT-qPCR analysis results clarified that UPP administration distinctly activated the IRS/PI3K/AKT signaling pathway, restrained PEPCK, G-6-Pase and Egr-1 genes, and affected the relative expression of HMGCR and LDLR genes. This study demonstrates that UPP could be applied as an adjuvant agent for the management of T2DM.

Identification and Screening of Potential Bioactive Peptides with Sleep-Enhancing Effects in Bovine Milk Casein Hydrolysate.

Casein tryptic hydrolysate (CTH) has been proven to possess stress-relieving and sleep-enhancing effects, but only one decapeptide YLGYLEQLLR (α-CZP) in CTH was reported to exhibit affinity for the benzodiazepine site of a GABAA receptor (GABAAR). This study aimed to compare the sleep-enhancing effects between CTH and α-CZP and to explore novel sleep-enhancing peptides. Our results showed that CTH significantly prolonged sleep duration in mice, which was almost 2-fold longer than that of α-CZP. The α-CZP in CTH was degraded more slowly than the synthetic α-CZP; meanwhile, CTH could release other potential sleep-enhancing peptides during gastrointestinal digestion. Additionally, two peptides YPVEPF and YFYPEL with strong sleep-enhancing activity were explored by virtual screening. Especially, YPVEPF could significantly prolong the sleep duration from 559.00 ± 272.24 to 2501.63 ± 1021.21 s and increase the sleep rate from 58.33 to 83.33% in mice. Moreover, YPVEPF and YFYPEL could bind with the Ser-205 and Phe-77 residues of GABAAR via hydrogen bonds and lipid contacts. They were largely released after digestion with 11.19 ± 0.15 and 1.78 ± 0.01 g/kg, respectively.