Comparative Study of the Effects of Dietary-Free and -Bound Nε-Carboxymethyllysine on Gut Microbiota and Intestinal Barrier.

Nε-carboxymethyllysine (CML) is produced by a nonenzymatic reaction between reducing sugar and ε-amino group of lysine in food and exists as free and bound forms with varying digestibility and absorption properties in vivo, causing diverse interactions with gut microbiota. The effects of different forms of dietary CML on the gut microbiota and intestinal barrier of mice were explored. Mice were exposed to free and bound CML for 12 weeks, and colonic morphology, gut microbiota, fecal short-chain fatty acids (SCFAs), intestinal barrier, and receptor for AGE (RAGE) signaling cascades were measured. The results indicated that dietary-free CML increased the relative abundance of SCFA-producing genera including Blautia, Faecalibacterium, Agathobacter, and Roseburia. In contrast, dietary-bound CML mainly increased the relative abundance of Akkermansia. Moreover, dietary-free and -bound CML promoted the gene and protein expression of zonula occludens-1 and claudin-1. Additionally, the intake of free and bound CML caused an upregulation of RAGE expression but did not activate downstream inflammatory pathways due to the upregulation of oligosaccharyl transferase complex protein 48 (AGER1) expression, indicating a delicate balance between protective and proinflammatory effects in vivo. Dietary-free and -bound CML could modulate the gut microbiota community and increase tight-junction expression, and dietary-free CML might exert a higher potential benefit on gut microbiota and SCFAs than dietary-bound CML.

Highland barley ß-glucan supplementation attenuated hepatic lipid accumulation in Western diet-induced non-alcoholic fatty liver disease mice by modulating gut microbiota.

Non-alcoholic fatty liver disease (NAFLD) has become one of the most common chronic liver diseases worldwide. NAFLD is caused by numerous factors, including the genetic susceptibility, oxidative stress, unhealthy diet, and gut microbiota dysbiosis. Among these, gut microbiota is a key factor and plays an important role in the development of NAFLD. Therefore, modulating the composition and structure of gut microbiota might provide a new intervention strategy for NAFLD. Highland barley ß-glucan (HBG) is a polysaccharide that can interact with gut microbiota after entering the lower gastrointestinal tract and subsequently improves NAFLD. Therefore, a Western diet was used to induce NAFLD in mouse models and the intervention effects and underlying molecular mechanisms of HBG on NAFLD mice based on gut microbiota were explored. The results indicated that HBG could regulate the composition of gut microbiota in NAFLD mice. In particular, HBG increased the abundance of short-chain fatty acids (SCFA)-producing bacteria (Prevotella-9, Bacteroides, and Roseburia) as well as SCFA contents. The increase in SCFA contents might activate the adenosine 5'-monophosphate (AMP)-activated protein kinase (AMPK) signaling pathway, thereby improving the liver lipid metabolism disorder and reducing liver lipid deposition.

Galactooligosaccharide or 2'-Fucosyllactose Modulates Gut Microbiota and Inhibits LPS/TLR4/NF-κB Signaling Pathway to Prevent DSS-Induced Colitis Aggravated by a High-Fructose Diet in Mice.

A high-fructose diet (HFrD) has been reported to exacerbate dextran sulfate sodium (DSS)-induced colitis. 2'-Fucosyllactose (FL) and galactooligosaccharide (GOS) have been shown, respectively, to have preventive and ameliorative effects on colitis, while limited research has explored whether GOS and FL may be equally protective or preventive in mice with HFrD. Here, we evaluated the protective effects of FL and GOS on colitis exacerbated by feeding HFrD and explored the underlying mechanisms. DSS-induced colitis was studied in four randomized C57BL/6J male mice (n = 8 mice/group). Among them, three groups were fed with HFrD, and two received either GOS or FL treatment, respectively. Gut microbial composition was analyzed by 16S rDNA gene sequencing. Intestinal barrier integrity and inflammatory pathway expression were measured using qPCR, immunofluorescence, and Western blot methods. Compared to the HFrD group, GOS or FL treatment increased the α-diversity of the gut microbiota, reduced the relative abundance of Akkermansia, and increased the content of short-chain fatty acids (SCFAs), respectively. Compared with the HFrD group, GOS or FL treatment improved the loss of goblet cells and the reduction of tight junction protein expression, thereby improving intestinal barrier integrity. Also, GOS or FL inhibited the LPS/TLR4/NF-κB signaling pathway and oxidative stress to suppress the inflammatory cascade compared with the HFrD group. These findings suggest that GOS or FL intake can alleviate HFrD-exacerbated colitis, with no significant difference observed between GOS and FL treatments.

Comparison of pharmacokinetics, biodistribution, and excretion of free and bound Nε-carboxymethyllysine in rats by HPLC-MS/MS.

As a representative product of advanced glycation end products, Nɛ-carboxymethyllysine (CML) exists in free and bound forms in vivo and in food with different bioavailability. To thoroughly understand the bioavailability of free Nɛ-carboxymethyllysine (CML) and bovine serum albumin (BSA)-CML in vivo after intragastric administration, pharmacokinetics, biodistribution, and excretion of CML in rats were investigated by high-performance liquid chromatography-tandem mass spectrometry (HPLC-MS/MS). Pharmacokinetics results revealed that free CML peaked at 1.83 h (1684.72 ± 78.08 ng/mL) and 1.33 h (1440.84 ± 72.48 ng/mL) in serum after intragastric administration of free CML and BSA-CML, demonstrating the higher absorption of free CML than BSA-CML. Besides, dietary free CML exhibited a relatively lower body clearance and tissue distribution than dietary BSA-CML based on the apparent volume of distribution and body clearance. Moreover, free CML was concentrated in the kidneys, indicating that kidneys were the target organ for the uptake of absorbed free CML. Additionally, the total excretion rate of CML in urine and feces were 37% and 60% after oral administration of free CML and BSA-CML. These results shed pivotal light on a better understanding of the biological effects of free and bound CML on health.

Comparison of metabolic fate, target organs, and microbiota interactions of free and bound dietary advanced glycation end products.

Increased intake of Western diets and ultra-processed foods is accompanied by increased intake of advanced glycation end products (AGEs). AGEs can be generated exogenously in the thermal processing of food and endogenously in the human body, which associated with various chronic diseases. In food, AGEs can be divided into free and bound forms, which differ in their bioavailability, digestion, absorption, gut microbial interactions and untargeted metabolites. We summarized the measurements and contents of free and bound AGE in foods. Moreover, the ingestion, digestion, absorption, excretion, gut microbiota interactions, and metabolites and metabolic pathways between free and bound AGEs based on animal and human studies were compared. Bound AGEs were predominant in most of the selected foods, while beer and soy sauce were rich in free AGEs. Only 10%-30% of AGEs were absorbed into the systemic circulation when orally administered. The excretion of ingested free and bound AGEs was approximately 90% and 60%, respectively. Dietary free CML has a detrimental effect on gut microbiota composition, while bound AGEs have both detrimental and beneficial impacts. Free and bound dietary AGEs changed amino acid metabolism, energy metabolism and carbohydrate metabolism. And besides, bound dietary AGEs altered vitamin metabolism, and glycerolipid metabolism.

Galactooligosaccharides ameliorate dietary advanced glycation end product-induced intestinal barrier damage in C57BL/6 mice by modulation of the intestinal microbiome.

Advanced glycation end products (AGEs) are increasingly recognized as potentially pathogenic components of processed foods, and long-term consumption of dietary AGEs triggers disruption of the intestinal barrier integrity and increases the risk of chronic diseases. Galactooligosaccharides (GOS) as prebiotics can modulate the intestinal microbiota and improve the intestinal barrier integrity. In this study, we aimed to investigate whether GOS could ameliorate the intestinal barrier damage induced by AGEs. The results showed an increased number of goblet cells (AGEs vs. H-GOS, 133.4 vs. 174.7, p < 0.05) and neutral mucin area (PAS positive area, 7.29% vs. 10.05%, p < 0.05). Upregulated expressions of occludin and claudin-1 and improved intestinal barrier integrity were observed in the H-GOS group. Using 16S rRNA sequencing analysis, we found that GOS significantly reduced the high enrichment of Akkermansia (16.95% vs. 1.29%, p < 0.05) induced by dietary AGEs while increasing the content of short-chain fatty acids. Fecal microbiota transplantation (FMT) showed that AGE-induced damage to the intestinal mucus barrier was reversed in the H-GOS transplanted group. Collectively, GOS ameliorated dietary AGE-induced intestinal barrier damage by reversing the dysregulated state of the intestinal microbiota. Our study lays the foundation for further research on dietary guidelines for populations with high AGE diets.

Highland barley ß-glucan alleviated western diet-induced non-alcoholic fatty liver disease via increasing energy expenditure and regulating bile acid metabolism in mice.

Non-alcoholic fatty liver disease (NAFLD) has become a public health burden. Controlling bile acids (BAs) metabolism and energy expenditure are  potential therapies for NAFLD. Because one of the main health effects of cereal ß-glucan (BG) is its ability to lower cholesterol by interacting with BAs, BG may regulate imbalances of the metabolism of BAs during NAFLD. Therefore, by using metabolic tests coupled with the profiling of hepatic BAs, we have assessed the effect of BG from highland barley on western diet (WD) induced NAFLD mice. BG treatment prevented fat accumulation and increased adipose lipolysis. These moderating effects were associated with an increased energy expenditure. Moreover, BG-treated mice enhanced the production of hepatic BAs, which may be connected with the activation of farnesoid X receptor (FXR) signaling in the liver and inhibition of FXR signaling in the ileum. Our results suggest that BG prevents fat accumulation by increasing energy expenditure, a mechanism associated with major changes in the composition of hepatic BAs.

Comparison of tissue distribution of free and protein bound Nɛ-carboxymethyllysine after long-term oral administration to mice.

Nɛ-Carboxymethyl-lysine (CML) is a primary advanced glycation end product that exists in the body and food as free and bound forms with different bioavailability and physiological effects. To compare the uptake, tissue distribution, and fecal excretion of dietary free and bound CML, free or bound CML were administered to healthy mice at 10 mg CML kg-1 body weight per day for 12 weeks. The results demonstrated that free CML was significantly absorbed in serum and accumulated in the colon, ileum, lung, kidneys, heart, spleen, brain, and liver after intake of free and bound CML, whereas no statistical increase was found in the accumulation of bound CML in the serum, lung, spleen, kidneys, and liver. The colon was the main tissue for the accumulation of free and total CML. Moreover, the accumulation of free CML in tissues and organs was significantly correlated with free CML levels in serum. In conclusion, consumption of bound CML caused a higher uptake, accumulation, and fecal excretion of CML in the body than intake of free CML.

Hypoglycemic Effects of Lycium barbarum Polysaccharide in Type 2 Diabetes Mellitus Mice via Modulating Gut Microbiota.

This study aims to explore the molecular mechanisms of Lycium barbarum polysaccharide (LBP) in alleviating type 2 diabetes through intestinal flora modulation. A high-fat diet (HFD) combined with streptozotocin (STZ) was applied to create a diabetic model. The results indicated that LBP effectively alleviated the symptoms of hyperglycemia, hyperlipidemia, and insulin resistance in diabetic mice. A high dosage of LBP exerted better hypoglycemic effects than low and medium dosages. In diabetic mice, LBP significantly boosted the activities of CAT, SOD, and GSH-Px and reduced inflammation. The analysis of 16S rDNA disclosed that LBP notably improved the composition of intestinal flora, increasing the relative abundance of Bacteroides, Ruminococcaceae_UCG-014, Intestinimonas, Mucispirillum, Ruminococcaceae_UCG-009 and decreasing the relative abundance of Allobaculum, Dubosiella, Romboutsia. LBP significantly improved the production of short-chain fatty acids (SCFAs) in diabetic mice, which corresponded to the increase in the beneficial genus. According to Spearman's correlation analysis, Cetobacterium, Streptococcus, Ralstonia. Cetobacterium, Ruminiclostridium, and Bifidobacterium correlated positively with insulin, whereas Cetobacterium, Millionella, Clostridium_sensu_stricto_1, Streptococcus, and Ruminococcaceae_UCG_009 correlated negatively with HOMA-IR, HDL-C, ALT, AST, TC, and lipopolysaccharide (LPS). These findings suggested that the mentioned genus may be beneficial to diabetic mice's hypoglycemia and hypolipidemia. The up-regulation of peptide YY (PYY), glucagon-like peptide-1 (GLP-1), and insulin were remarkably reversed by LBP in diabetic mice. The real-time PCR (RT-PCR) analysis illustrated that LBP distinctly regulated the glucose metabolism of diabetic mice by activating the IRS/PI3K/Akt signal pathway. These results indicated that LBP effectively alleviated the hyperglycemia and hyperlipidemia of diabetic mice by modulating intestinal flora.

A "two-step" assay based on electro-activation for rapid determination of methylglyoxal in honey and beer.

Methylglyoxal (MGO), a dicarbonyl compound in living organism, food and environment, has been associated with disease diagnosis and human health. The current electrochemical detection methods rely on the use of advanced materials. In this work, a non-advanced materials "two-step" assay including electrode electro-activation and MGO detection was developed. In the section of electro-activation, an activation method of GCE for MGO detection was established; and the composition changes on GCE surface caused by electro-activation, including functional groups and surface defects, have been carefully studied. The effect of carbonyl and surface defects induced by electro-activation on MGO detection was discussed. In section of MGO detection, the raise of background current caused by electro-activation was minimized by background subtraction; and the effect of interferences can be weakened by adjusting pH. The MGO signal on proposed activated GCE improved 20-fold than bare GCE. The recoveries were 72.38-109.16% in honey and beer, and RSDs were 0.24-9.63% without significant difference with HPLC method and comparable with advanced material modified sensors.

Hepatic Lipidomics Analysis Reveals the Ameliorative Effects of Highland Barley ß-Glucan on Western Diet-Induced Nonalcoholic Fatty Liver Disease Mice.

Nonalcoholic fatty liver disease (NAFLD) is characterized by marked imbalances in lipid storage and metabolism. Because the beneficial health effects of cereal ß-glucan (BG) include lowering cholesterol and regulating lipid metabolism, BG may alleviate the imbalances in lipid metabolism observed during NAFLD. The aim of our study was to investigate whether BG from highland barley has an effect on western diet-induced NAFLD in mice. Using lipidomics, we investigated the underlying mechanisms of BG intervention, and identified potential lipid biomarkers. The results reveal that BG (300 mg/kg body weight) significantly alleviated liver steatosis. Lipidomics analysis demonstrated that BG also altered lipid metabolic patterns. We were able to identify 13 differentially regulated lipid species that may be useful as lipid biomarkers. Several genes in the hepatic lipid and cholesterol metabolism pathways were also modulated. These findings provide evidence that BG ameliorates NAFLD by altering liver lipid metabolites and regulating lipid metabolism-related genes.

A facile, inexpensive and green electrochemical sensor for sensitive detection of imidacloprid residue in rice using activated electrodes.

The development of sensitive, facile, cost-effective and eco-friendly sensors is essential for monitoring imidacloprid (IDP) residue on a large scale. Compared with popular modification of electrodes with advanced materials, electrochemical activation is promising at this point. In this paper, we found that strongly basic electrolytes (e.g. KOH and K3PO4) and applying cyclic potential during the activating process are beneficial to greatly amplify the electro-reduction response of IDP by nearly 16 times. Combining the characterization of activated electrodes with electrochemical behavior analysis of IDP, it is speculated that specific oxygen-contained functional groups were formed to bond with IDP molecules, leading to fast electron transfer kinetics. Then a sensitive IDP sensor has been developed with a low limit of detection (LOD) of 0.03 µM in the range of 0.1-100 µM. The methodological evaluation including reproducibility, stability and recovery has been also carefully studied, verifying the potential of proposed activated electrodes for application in rice samples.

Correction: Anthocyanins from the fruits of Lycium ruthenicum Murray improve high-fat diet-induced insulin resistance by ameliorating inflammation and oxidative stress in mice.

Correction for 'Anthocyanins from the fruits of Lycium ruthenicum Murray improve high-fat diet-induced insulin resistance by ameliorating inflammation and oxidative stress in mice' by Baoming Tian et al., Food Funct., 2021, 12, 3855-3871, DOI: 10.1039/D0FO02936J.

Anthocyanins from the fruits of Lycium ruthenicum Murray improve high-fat diet-induced insulin resistance by ameliorating inflammation and oxidative stress in mice.

A high-fat diet (HFD) promotes tissue inflammation, oxidative stress and insulin resistance (IR), thereby contributing to the development of obesity and diabetes. Anthocyanins from Lycium ruthenicum (AC) have demonstrated anti-obesity effects and modulated IR. To investigate the mechanism by which AC attenuates the adverse effects of consuming a HFD, C57BL/6J mice were fed a HFD supplemented with AC or a control diet without AC for 12 weeks. AC supplementation decreased the amount of weight gain, hepatic lipid, and sequentially improved dyslipidemia, inflammation, oxidative stress, and IR in HFD-fed mice. Molecular data revealed that AC inhibited hepatic inflammation by reducing TLR4/NF-κB/JNK in the liver tissues and ameliorated oxidative stress by activating the Nrf2/HO-1/NQO1 pathway. Thus, AC might activate IRS-1/AKT and prevent HFD-induced gluconeogenesis and IR by ameliorating inflammation and oxidative stress. Modulation of inflammation and oxidative stress with AC may represent a promising target for the treatment of IR and provide insight into the mechanism by which AC protects against obesity.

Lycium ruthenicum Anthocyanins Attenuate High-Fat Diet-Induced Colonic Barrier Dysfunction and Inflammation in Mice by Modulating the Gut Microbiota.

SCOPE: Gut barrier dysfunction and inflammation originating from a dysbiotic gut microbiota (GM) are strongly associated with a high-fat diet (HFD). Anthocyanins from Lycium ruthenicum (ACs) show antiobesity effects through modulating the GM. However, the mechanism linking the antiobesity effects of ACs and GM modulation remains obscure. METHODS AND RESULTS: To investigate the ameliorative effects of ACs on colonic barrier dysfunction and inflammation, mice are fed an HFD with or without ACs at doses of 50, 100, and 200 mg kg-1 for 12 weeks. AC supplementation reduced weight gain, enriched short-chain fatty acid (SCFA)-producing bacteria (e.g., Ruminococcaceae, Muribaculaceae, Akkermansia, Ruminococcaceae_UCG-014, and Bacteroides) and SCFA content, depleted endotoxin-producing bacteria (e.g., Helicobacter and Desulfovibrionaceae), and decreased endotoxin (i.e., lipopolysaccharide) levels. SCFAs substantially activated G protein-coupled receptors (GPRs), inhibited histone deacetylases (HDAC), increased intestinal tight junction mRNA and protein expression levels, reduced intestinal permeability, and protected intestinal barrier integrity in HFD-induced mice. These effects mitigate intestinal inflammation by inhibiting the LPS/NF-κB/TLR4 pathway. CONCLUSION: These data indicates that ACs can mitigate colonic barrier dysfunction and inflammation, induce SCFA production and inhibit endotoxin production by modulating the GM in HFD-fed mice. This finding provides a clue for understanding the antiobesity effects of ACs.

Health beneficial effects of resistant starch on diabetes and obesity via regulation of gut microbiota: a review.

Resistant starch (RS) is well known to prevent type 2 diabetes mellitus (T2DM) and obesity. Recently, attention has been paid to gut microbiota which mediates the RS's impact on T2DM and obesity, while a mechanistic understanding of how RS prevents T2DM and obesity through gut microbiota is not clear yet. Therefore, this review aims at exploring the underlying mechanisms of it. RS prevents T2DM and obesity through gut microbiota by modifying selective microbial composition to produce starch-degrading enzymes, promoting the production of intestinal metabolites, and improving gut barrier function. Therefore, RS possessing good functional features can be used to increase the fiber content of healthier food. Furthermore, achieving highly selective effects on gut microbiota based on the slight differences of RS's chemical structure and focusing on the effects of RS on strain-levels are essential to manipulate the microbiota for human health.

Microbiome-Metabolomics Analysis of the Impacts of Long-Term Dietary Advanced-Glycation-End-Product Consumption on C57BL/6 Mouse Fecal Microbiota and Metabolites.

Thermally processed diets are widely consumed, although advanced-glycation end products (AGEs) are unavoidably formed. AGEs, clusters of protein-cross-linking products, become less digestible because they impair intestinal peptidase proteolysis. We characterized the impacts of dietary AGEs on gut microbiota through a microbiome-to-metabolome association study. C57BL/6 mice were fed a heat-treated diet (high-AGE diet, H-AGE) or a standard AIN-93G diet (low-AGE diet, L-AGE) for 8 months. Fecal-microbiota composition was examined by 16S rDNA sequencing, and fecal-metabolome profile was evaluated by gas chromatography-tandem time-of-flight mass spectrometry (GC-TOF-MS). Reduced α-diversity and altered microbiota composition with elevated Helicobacter levels were found in the H-AGE group, and among the 57 perturbed metabolites, protein-fermentation products (i.e., p-cresol and putrescine) were increased. Major dysfunctional metabolic pathways were associated with carbohydrate and amino acid metabolism in two groups. Moreover, high correlations were found between fluctuant gut microbiota and metabolites. These findings might reveal the underlying mechanisms of the detrimental impacts of dietary AGEs on host health.

Dietary advanced glycation end products modify gut microbial composition and partially increase colon permeability in rats.

SCOPE: The adverse impacts of dietary advanced glycation end products (AGEs) on health are currently of interest. These compounds are inevitably formed during thermal food processing and make foods less digestible because of protein cross-linking. This study examined not only whether dietary AGEs alter cecal microbiota and their metabolites but also their effects on colon permeability. METHODS AND RESULTS: Sprague-Dawley rats were exposed to a high-AGEs diet (AGEs content was increased by heating food at 125°C/3 h) for 6, 12, or 18 weeks. Cecal microbiota was analyzed by 16S rDNA gene sequencing. Colon permeability was assessed through histopathology, immunohistochemistry and endotoxin testing. Microbial metabolites (e.g. ammonia and short-chain fatty acids (SCFAs)) were also measured. AGEs treatment reduced the diversity and richness of the microbiota, especially saccharolytic bacteria such as Ruminococcaceae and Alloprevotella, which can produce SCFAs, whereas some putatively harmful bacteria (Desulfovibrio and Bacteroides) were increased. Protein fermentation was enhanced, supported by elevated ammonia and branched-chain fatty acid levels (p < 0.05). Additionally, the colonocytes structure changed and the expression of tight junction proteins in colon were decreased. CONCLUSION: Dietary AGEs detrimentally modulate gut microbial ecology and may partially increase colon permeability, which can adversely impact host health.