Intestinal Cells Research Articles

Effects of Microplastic on the Gastrointestinal Tract and Gut microbiome of Sprague Dawley Rats

ABSTRACT Nowadays there is evidence that animals are exposed to microplastics (MPs) via food, however there is not much known about their effect on tissue of gastrointestinal tract and on gut microbiome. In this study, adult male rats were fed with polyethylene MPs, mixed with standard pellet food for 24 days at different concentrations ranging from 0 to 1000 μg/day. At the end of experiment, rats were euthanized and histopathological investigation using light and electron microscopy and the gut microbiome was analyzed using MiSeq 16S rRNA amplicon sequencing. Microscopic analysis revealed significant impact of MPs on the intestine, including visible changes to the crypt area as well as reduction in mucus secretion. Autophagic vacuoles were observed in the livers of the rats fed with MPs at 100 µg/day. Oxidative stress was evident in rats fed with 1000 µg/day of MPs, as indicated by the presence of myeloid bodies in both intestinal and hepatic cells. The gut microbiome was also affected by MPs. Although, no distinct clusters were formed when the bacterial communities in the different treatments were ordinated by multidimensional scaling (NMDS), the diversity indices including operational taxonomic unit (OTU) richness and Chao1 exhibited an increasing trend with increasing MPs concentration. Pearson correlations revealed a linear increase in the relative sequence abundance of Clostridia (R=0.036, P=0.003) with increasing MPs concentration, but not in the case of other bacterial groups,. Lactobacillus faecis was the most abundant OTU in the entire dataset, and its relative abundance decreased significantly with increasing MPs concentration. We conclude that exposure to MPs can disrupt cellular function and disturb microbiome balance, and in the event of prolonged exposure, organisms might experience unpredictable effects.

PSVII-16 Recombinant bovine lactoferrin expressed in Schizochytrium sp. alleviates inflammation and oxidative damage in calf intestinal epithelial cells

Abstract This study aims to generate recombinant bovine lactoferrin (rbLF) by expressing it in Schizochytrium sp. (SZ) and evaluate its effects on calf intestinal epithelial cells (CIECs) exposed to lipopolysaccharide (LPS) or hydrogen peroxide-induced damage. The bovine lactoferrin expression cassette was transfected into Schizochytrium sp. via 18s rRNA gene recombination. Primary CIECs were utilized in this investigation, and the optimal dose of rbLF was evaluated by cell counting kit-8 assay after being treated with different concentrations of rbLF for 24 h. The optimal dose of rbLF was then used as a medium supplement before or after 24 h of 80 μg/mL LPS challenge or 4 h of 400 μM of hydrogen peroxide (H2O2) challenge. Cell viability, gene expression of tumor necrosis factor (TNF) -α, toll-like receptor (TLR) -4, interleukin (IL) -17A, zonula occludens-1 (ZO-1), and claudin-1, as well as protein levels of TNF-α, IL-17A, ZO-1, and claudin-1, were assessed to elucidate the preventive and therapeutic effects of rbLF. Data analysis was performed using SAS 9.4 by F-test, The statistical significance was declared at P < 0.05. Results indicate that rbLF demonstrated no cytotoxicity within the 50 to 1,000 μg/mL range on CIECs. Treatment with 175 μg/mL rbLF for 24 h increased cell viability, expression, and protein concentrations of ZO-1 and claudin following hydrogen peroxide challenge (P < 0.05). Moreover, rbLF pretreatment for 24 hours, despite decreasing cell viability after the H2O2 challenge, increased the expression and protein concentrations of ZO-1 and claudin-1 compared with non-rbLF-treated groups (P < 0.05). When used to prevent LPS-induced inflammation, rbLF enhanced the gene expression and protein concentrations of TNF-α, ZO-1, and claudin-1 compared with controls (P < 0.05). As a treatment for LPS-induced inflammation, rbLF decreased TLR4, TNF-α, and IL-17A expression levels (P < 0.05). These findings suggest that SZ-expressed rbLF is safe for calve intestinal cells and 175 μg/mL of rbLF may mitigate inflammation-induced intestinal barrier dysfunction.

Toxicity Evaluation of Potassium Sorbate In Vivo with Drosophila Melanogaster.

Potassium sorbate (PS) is a preservative widely used in the food, pharmaceutical, and cosmetics industries. Improper and careless use of PS can lead to various health issues and potential environmental problems. Drosophila is capable of making rapid and sensitive responses to stress or other stimuli. Here we utilized Drosophila as a model organism to evaluate the potential toxicity of PS. Our study revealed that PS ingestion reduced the lifespan and fecundity of Drosophila. In addition, excessive PS ingestion led to cell apoptosis and ROS accumulation in the midgut. Furthermore, PS intake also enhanced the mitophagy of midgut cells. Strikingly, PS affected the cell differentiation progression as well, leading to the production of more enteroendocrine (EE) cells. We further demonstrated that the expression of notch (N), a vital player in intestinal stem cell (ISC) differentiation, was down-regulated in the midgut. This indicates that the differentiation progression was affected potentially by repressing the N expression.

Establishment of an in-vitro inflammatory bowel disease model using immunological differentiation of Caco-2 cells

Studies on intestinal cell differentiation, particularly in dextran sodium sulfate (DSS)-induced inflammatory bowel disease (IBD), have predominantly focused on the disruption of intestinal crypts and suppressive effects on the intestinal microbiota; however, repeated administration of DSS is required to induce inflammatory pathology, and there is a lack of observation of early responses and consideration of differentiation stages. Although colonic adenocarcinoma (Caco-2) cells can be used as intestinal cell models, research on these cells in an immature state is limited. We, therefore, investigated the relationship between Caco-2 cell culture duration and immunological differentiation using α-defensin5 (DEFA5) as an indicator of intestinal immunity and differentiation. Changes in protein and gene expression levels in response to DSS were examined at each differentiation stage. Expression of immune- and differentiation-related proteins, including DEFA5 and lysozyme, was evident from Day 8 of culture. Immune responses to DSS varied with the differentiation stage, affecting cell viability and cytokine expression.•Caco-2 cell culture duration correlates with the differentiation stage of Paneth cells.•DSS exposure elicits different effects depending on the differentiation stage.•Our in-vitro model of IBD facilitates the characterization of the cell differentiation process and provides a methodology to help elucidate the causal mechanisms of IBD.

236 Increased dietary protein supports key metabolic and intestinal tissue functions in weaned pigs, without increasing gastrointestinal pathogen abundance

Abstract A common dietary strategy to prevent diarrhea in weaned pigs is feeding low protein diets in the period immediately following weaning. The goal of this intervention is to decrease dietary protein sufficiently to reduce pathogen proliferation and protein fermentation without negatively impacting growth performance. One potential alternative to low protein diets is provision of dietary fiber. However, many studies examining the role of protein and fiber on gastrointestinal microbiota and post-weaning diarrhea are complicated by the presence of other substrates, including polyphenols and antinutritional factors in complex ingredients. To assess the role of increased protein and fiber on gut microbiota and host metabolism, semi-purified diets differing in crude protein (CP) and crude fiber (CF) were fed to weaned pigs (n = 40; 28 d of age). Diets with high protein (HP; CP 24.3%, CF 6.4%), high fiber (HF; CP 18.9%, CF 9.4%), or both (HFHP; CP 24.9%, CF 9.5%) were compared with a control (CON; CP 18.1%, CF 6.3%) diet with industry standard crude protein and fiber levels (n = 10/group). Growth performance and diarrhea prevalence were measured alongside gut microbiota composition and metabolites. Pigs fed the HP and HFHP diets had significantly improved feed efficiency compared with both CON and HF diets. Increased protein fermentation, measured through production of biogenic amines, was observed in HP fed animals (P < 0.001). These effects were mitigated by addition of fiber in the HFHP fed pigs. Despite increased markers of protein fermentation, post-weaning diarrhea incidence was not greater in HP fed animals and no increases in gastrointestinal pathogen abundance were detected. Metabolomic and transcriptomic analyses identified critical alterations in host metabolism and cecal transcriptome in the CON compared with the HP fed animals. While all diets were formulated to meet the dietary requirements of weaned pigs, diets with lower protein levels (CON and HF) induced alteration in transcripts from the serine synthesis pathways (P < 0.001) and integrated stress response (P < 0.001) in cecal tissue alongside increases in metabolic pathways related to lysine degradation (P = 0.005). These results challenge the current practice of low protein feeding following weaning by demonstrating a detrimental effect of low protein diets on intestinal cell function and muscle accretion. This suggests that with careful ingredient selection, increased dietary protein post-weaning could better support pig health and performance compared with current industry standard diets.

Effects of pasteurized Akkermansia muciniphila on lipid metabolism disorders induced by high-fat diet in zebrafish (Danio rerio)

Akkermansia muciniphila (Akk) is an important probiotic that can help alleviate metabolic disorders. Recent research indicates that pasteurized Akkermansia muciniphila (P-Akk) is even more effective. To investigate the effect of P-Akk on improving lipid metabolism disorders caused by a high-fat diet (HFD) in fish, zebrafish were fed two HFDs containing 0 (HFD group, as control) and 1 × 109 CFU/g P-Akk (P-Akk group) for 28 days. The addition of P-Akk significantly decreased the number of lipid droplets in the liver and improved the mitochondrial structure in the intestine, while also altering gene expression related to lipid metabolism in HFD-fed zebrafish. Furthermore, intestinal transcriptomic network analysis and results from primary intestinal cells indicated that P-Akk may enhance mitochondrial function by activating the AMPK/PGC1α pathway. Although P-Akk did not significantly alter the composition of the gut microbiota, it did increase the steady-state index of intestinal microbiota. Moreover, P-Akk upregulated the abundance of beneficial probiotics, (Plesiomonas, Cetobacterium, and Bacillus), while downregulating the abundance of pathogens (Aeromonas, Pseudomonas, Acinetobacter, and Streptococcus). Collectively, these findings indicate that P-Akk has the potential to serve as a green and safe feed additive for improving HFD-induced lipid metabolism disorders in fish.

515 Effects of Bacillus licheniformis HG76 on barrier function and inflammation in the intestinal epithelia cells and gut health of weaned piglets under enterotoxigenic Escherichia coli F4 infection

Abstract Enterotoxigenic Escherichia coli (ETEC) F4 secretes enterotoxins and invades the small intestine, which damages the gut barrier function and stimulates the immune reactions characterized by inflammation. In the current study, the effects of B. licheniformis HG76 on gut health were evaluated with ETEC F4-induced IPEC-J2 cells and weaned piglets. The B. licheniformis HG76 was isolated from cecal contents of Rhode Island Red laying hens raised without using antibiotics. The results from the in vitro study showed that the ETEC F4 infection significantly induced IPEC-J2 cell death, and this effect was significantly attenuated by B. licheniformis HG76 treatment (P < 0.05). The gene expressions of zona occludens (ZO1) and tumor necrosis factor (TNF)-α were significantly increased during ETEC F4 infection with B. licheniformis HG76 treatment (P < 0.05). Meanwhile, the ETEC F4 infection significantly increased the mRNA abundance of toll-like receptors (TLR2, TLR4, and TLR5) (P < 0.05), but the B. licheniformis HG76 treatment alleviated this increase. For the in vivo study, 28 weaned piglets susceptible to ETEC F4 at 28 d of age were individually allotted to four experimental groups: NNC: unchallenged control group; NC: challenged control group; PC: antibiotic group (80 mg·kg-1 of avilamycin); and HG76: Bacillus group (1.0 × 109 CFU kg-1 diet of B. licheniformis HG76). The in vivo results showed during the post-challenge phase, ETEC F4 infection decreased average daily gain (ADG) and increased the severity of diarrhea at 44 h post-inoculum (hpi), respectively, in the NC group compared with the NNC group (P < 0.05). In addition, the infection decreased the villus height (VH) in the duodenum of pigs from the NC, PC and HG76 groups compared with the NNC group. The VH of jejunum and ileum in the pigs from the HG76 group were higher than other groups (P < 0.05). There was an increase (P < 0.05) of crypt depth (CD) in the jejunum and ileum of pigs from the HG76 group compared with those from the NNC group. The pigs in the NNC group had the greatest duodenal VH:CD ratio, but the least ileal VH:CD ratio among all the treatment groups. The Vmax value of sucrase in the HG76 group was greater than the NNC group (P < 0.05). When compared with the NC group, the relative mRNA abundance of interleukin 6 (IL6) was increased in the PC group (P < 0.05). In summary, these results suggest that B. licheniformis HG76 can protect the intestinal cells from ETEC F4 infection by inhibiting inflammation and maintaining barrier integrity, and the supplementation of B. licheniformis HG76 can improve the gut development and increase the digestive enzyme activity of ETEC F4 challenged piglets.

Critical Suitability Evaluation of Caco-2 Cells for Gut-on-a-Chip.

Currently, culturing Caco-2 cells in a Gut-on-a-chip (GOC) is well-accepted for developing intestinal disease models and drug screening. However, Caco-2 cells were found to overexpress surface proteins (e.g., P-gp) compared with the normal intestinal epithelial cells in vivo. To critically evaluate the challenge and suitability of Caco-2 cells, a GOC integrated with a carcinoembryonic antigen (CEA) biosensor was developed. This three-electrode system electrochemical sensor detects CEA by antigen-antibody specific binding, and it exhibits high selectivity, excellent stability, and good reproducibility. Under dynamic culturing in the GOC, Caco-2 cells exhibited an intestinal villus-like structure and maintained tissue barrier integrity. Meanwhile, CEA was discovered to be secreted from 0 to 0.22 ng/mL during the 10-day culturing of Caco-2 cells. Especially, CEA secretion increased significantly with the differentiation of Caco-2 cells after 6 days of culturing. The sustained high-level CEA secretion may induce cells to avoid apoptotic stimuli, which faithfully reflects the efficacy of a new drug and the mechanism of intestinal disease. Different kinds of cell types (e.g., intestinal primary cells, stem cell-induced differentiation) in the GOC should be attempted for drug screening in the future.

Neonatal Fc receptor is a functional receptor for classical human astrovirus.

Human astrovirus (HAstV) is a global cause of gastroenteritis in infants, the elderly, and the immunocompromised. However, the molecular mechanisms that control its susceptibility are not fully understood, as the functional receptor used by the virus has yet to be identified. Here, a genome-wide CRISPR-Cas9 library screen in Caco2 cells revealed that the neonatal Fc receptor (FcRn) can function as a receptor for classical HAstV (Mamastrovirus genotype 1). Deletion of FCGRT or B2M, which encode subunits of FcRn, rendered Caco2 cells and intestinal organoid cells resistant to HAstV infection. We also showed that human FcRn expression renders non-susceptible cells permissive to viral infection and that FcRn binds directly to the HAstV spike protein. Therefore, our findings provide insight into the entry mechanism of HAstV into susceptible cells. We anticipate that this information can be used to develop new therapies targeting human astroviruses, providing new strategies to treat this global health issue.

Trained immunity of intestinal tuft cells during infancy enhances host defense against enteroviral infections in mice

Innate immune cells have been acknowledged as trainable in recent years. While intestinal tuft cells are recognized for their crucial roles in the host defense against intestinal pathogens, there remains uncertainty regarding their trainability. Enterovirus 71 (EV71), a prevalent enterovirus that primarily infects children but rarely infects adults. At present, there is a significant expansion of intestinal tuft cells in the EV71-infected mouse model, which is associated with EV71-induced interleukin-25 (IL-25) production. Further, we found that IL-25 pre-treatment at 2 weeks old mouse enabled tuft cells to acquire immune memory. This was evidenced by the rapid expansion and stronger response of IL-25-trained tuft cells in response to EV71 infection at 6 weeks old, surpassing the reactivity of naïve tuft cells in mice without IL-25-trained progress. Interestingly, IL-25-trained intestinal tuft cells exhibit anti-enteroviral effect via producing a higher level of IL-25. Mechanically, IL-25 treatment upregulates spermidine/spermine acetyl-transferase enzyme (SAT1) expression, mediates intracellular polyamine deficiency, further inhibits enterovirus replication. In summary, tuft cells can be trained by IL-25, which supports faster and higher level IL-25 production in response to EV71 infection and further exhibits anti-enteroviral effect via SAT1-mediated intracellular polyamine deficiency. Given that IL-25 can be induced by multiple gut microbes during human growth and development, including shifts in gut flora abundance, which may partially explain the different susceptibility to enteroviral infections between adults and children.

Maternal High-Fat Diet Exacerbates Epicutaneous Sensitization and Oral Challenge-Induced Food Allergy to Ovalbumin in Offspring Mice.

Dietary factors have been associated with an increased prevalence of food allergy (FA). However, little is known about how an unhealthy diet in early life affects FA reactions in offspring. The objective of this study is to provide a scientific foundation for developing and promoting healthy dietary patterns in early life. In this study, we found that maternal high-fat diet (HFD) during pregnancy and lactation exacerbates FA (HFD-FA) in offspring mice, leading to increased serum levels of mast cell protease 1. First, we studied the systemic immunity of the HFD-FA mice and observed elevated levels of proinflammatory cytokines (IL-4, IL-6, and IL-1β) and a reduced frequency of Treg cells in splenocytes. Additionally, the HFD-FA mice showed increased gut permeability, accumulation of intestinal mast cells, and a decrease in the Treg cell frequency in the mesenteric lymph nodes. Furthermore, our findings also indicated a reduction in gut microbial diversity and abundance in HFD-FA mice. Importantly, lipid metabolism profiling revealed unique lipid profiles in the HFD-FA mice, with significant upregulation of triglycerides and downregulation of sphingolipids. Taken together, our results suggest that maternal HFD alters intestinal homeostasis and increases FA susceptibility in offspring mice.

Segmental patterning of microbiota and immune cells in the murine intestinal tract

ABSTRACT The intestine exhibits distinct characteristics along its length, with a substantial immune cell reservoir and diverse microbiota crucial for maintaining health. This study investigates how anatomical location and regional microbiota influence intestinal immune cell abundance. Using conventionally colonized and germ-free mice, segment-specific immune cell composition and microbial communities were assessed. Metagenomic sequencing analyzed microbiome variations, while flow cytometry and immunofluorescence examined immune cell composition. Microbiome composition varied significantly along the intestine, with diversity and abundance increasing from upper to lower segments. Immune cells showed distinct segment-specific patterning influenced by microbial colonization and localization. T cell subsets displayed varied dependence on microbiome presence and anatomical location. This study highlights locoregional differences in intestinal immune cell and microbiome composition, identifying immune subsets susceptible to microbiota presence. The findings provide context for understanding immune cell alterations in disease models.

A Comprehensive Review of the Ethnobotanical Uses, Pharmacological, and Toxicological Profiles of Piper capense L.f. (Piperaceae)

Commonly known as wild pepper, Piper capense (P. capense) is a culinary herb mainly used as a secret in preparation of “Nkui” and “Nah poh” in Bayangam, West Cameroon. However, it also has many interesting pharmacological properties, which is why the people of sub-Saharan Africa so highly prize it for the treatment of multiple human pathologies. This study aimed to highlight the traditional uses, phytochemical composition, biological activities, and toxicological profile of the P. capense plant, to draw the attention of pharmaceutical companies to its enormous potential for the development of future phyto- or pharmaceutical products. Documentary research was meticulously carried out in the Web of Sciences, Scopus, Pubmed/Medline, and Google Scholar databases according to PRISMA 2020 guidelines. The results show that extracts and compounds isolated from Piper capense have interesting anticancer, antibacterial, antimalarial, hypoglycemic, anti-epileptic, and antidepressant activities. Methanolic extracts and essential oils from P. capense exhibit no harmful effects when directly applied to normal human hepatocytes, umbilical cord cells, intestinal cells, and keratinocyte cell lines. Additionally, methanolic extracts administered acutely or subchronically at low doses (≤250 mg/kg body weight) in Wistar rats also demonstrate no adverse effects. In conclusion, given its interesting activities, P. capense is a viable option for developing new antimalarial, anticancer, antibacterial, hypoglycemic, anti-epileptic, and antidepressant drugs. However, many avenues still need to be explored before translation into drugs.

Increased levels of anti-Encephalitozoon intestinalis antibodies in patients with colorectal cancer.

The prevalence of microsporidiosis in the general population, or within specific groups of individuals/patients, is largely underestimated. The absence of specific seroprevalence tools limits knowledge of the epidemiology of these opportunistic pathogens, although known since the 1980s. Since microsporidia hijack the machinery of its host cell and certain species multiply within intestinal cells, a potential link between the parasite and colorectal cancer (CRC) has been suggested. To explore a potential epidemiological link between microsporidia and CRC, we evaluated the seroprevalence of Encephalitozoon intestinalis among CRC patients and healthy subjects using ELISA assays based on two recombinant proteins, namely rEiPTP1 and rEiSWP1, targeting polar tube and spore wall proteins. ELISA were performed in 141 CRC patients and 135 healthy controls. Patients with CRC had significantly higher anti-rEiPTP1 IgG levels than subjects in the control group. Anti-rEiPTP1 IgG, anti-rEiSWP1 IgG and anti-rEiPTP1 IgA levels were significantly increased among men with CRC compared to healthy men. Women with CRC who had died had higher rEiSWP1 IgG levels than those who were still alive. These higher antibody levels against microsporidia in patients with CRC suggest a relationship between microsporidia and pathophysiology of CRC.

Molecular cross-talk among human intestinal bifidobacteria as explored by a human gut model.

Bifidobacteria are well known as common and abundant colonizers of the human gut and are able to exert multiple beneficial effects on their host, although the cooperative and competitive relationships that may occur among bifidobacterial strains are still poorly investigated. Therefore, to dissect possible molecular interactions among bifidobacterial species that typically colonize the human gut, three previously identified bifidobacterial prototypes, i.e., B. bifidum PRL2010, B. breve PRL2012, and B. longum PRL2022 were cultivated individually as well as in bi- and tri-association in a human gut-simulating medium. Transcriptomic analyses of these co-associations revealed up-regulation of genes predicted to be involved in the production of extracellular structures including pili (i.e., flp pilus assembly TadE protein gene), exopolysaccharides (i.e., GtrA family protein gene) and teichoic acids (i.e., ABC transporter permease), along with carbohydrate, amino acid and vitamin metabolism-related genes (i.e., exo-alpha-sialidase; beta-galactosidase and pyridoxamine kinase), suggesting that co-cultivation of bifidobacteria induces a response, in individual bifidobacterial strains, aimed at enhancing their proliferation and survival, as well as their ability to cooperate with their host to promote their persistence. Furthermore, exposure of the selected prototypes to human cell line monolayers unveiled the ability of the bifidobacterial tri-association to communicate with their host by increasing the expression of genes involved in adherence to/interaction with intestinal human cells. Lastly, bifidobacterial tri-association promoted the transcriptional upregulation of genes responsible for maintaining the integrity and homeostasis of the intestinal epithelial barrier.

Antagonistic insulin-like signaling influencing glucose regulation in C. elegans

The insulin/ IGF-signaling (IIS) is an evolutionary conserved signaling pathway within eukaryotic organisms that regulates glucose metabolism and cell division. The nematode, C. elegans is an excellent model for exploring IIS as they have 40 insulin-like peptides that act on a single receptor, DAF-2. We aim to explore the relationships between IIS and glucose metabolism, focusing on the antagonistic insulin-like peptide, INS-17 and its influence over FDGT-1 (facilitated glucose transporter 1). FDGT-1 is expressed throughout the nematode, including the basolateral membrane of intestinal cells and where it is theorized to export glucose to the surrounding somatic cells rather than absorb it from the intestinal tract. We hypothesize that IIS controls transcription of FDGT-1 and antagonistic peptides should lead to a decrease in FDGT-1 transcription, resulting in the increased lifespan and increased fat accumulation observed in daf-2(lf) loss of function and ins-17(oe) over expressing strains. In this study, survival assays are used to assess the impact which high-glucose content has on lifespan, along with Oil Red O (ORO) lipid staining to visualize lipid accumulation amongst conditions and protein quantification of FDGT-1:GFP to evaluate FDGT-1 protein levels. Our findings are consistent with the idea of low insulin signaling leading to extended lifespan as daf-2(lf) lived significantly longer than WT in both 0% and 2% glucose conditions and ins-17(oe) living significantly longer than wildtype in 2% glucose conditions. The ORO lipid staining revealed that high glucose conditions and low insulin signaling result in lipid accumulation within the nematode. Using the fdgt-1:gfp strain, we were able to determine an increase in the FDGT-1 in the presence of glucose, suggesting FDGT-1 expression is influenced by glucose. These findings aim to further our understanding of IIS regulating glucose uptake and metabolism, along with the significance of the FDGT-1 glucose transporter.

Pyrroloquinoline Quinone Alleviates Intestinal Inflammation and Cell Apoptosis via the MKK3/6-P38 Pathway in a Piglet Model.

This study investigates the underlying mechanism through which dietary supplementation of pyrroloquinoline quinone disodium (PQQ) alleviates intestinal inflammation and cell apoptosis in piglets challenged with lipopolysaccharide (LPS). Seventy-two barrows were divided into three groups: control (CTRL), LPS challenged (LPS), and LPS challenged with PQQ supplementation (PQQ + LPS). On d 7, 11, and 14, piglets received intraperitoneal injections of LPS or 0.9% of NaCl (80 μg/kg). After a 4 h interval following the final LPS injection on d 14, blood samples were obtained, and all piglets were euthanized for harvesting jejunal samples. The results showed that dietary supplementation of PQQ improved the damage of intestinal morphology, increased the down-regulated tight junction proteins, and reduced the increase of serum diamine oxidase activity, the intestinal fatty acid binding protein, and TNF-α levels in piglets challenged with LPS (p < 0.05). The proteomics analysis revealed a total of 141 differentially expressed proteins (DEPs), consisting of 64 up-regulated DEPs and 77 down-regulated DEPs in the PQQ + LPS group compared to the LPS group. The KEGG pathway analysis indicated enrichment of the tight junction pathway and the apoptosis pathway (p < 0.05). Compared to the LPS group, the piglets in the PQQ + LPS group had increased levels of Bcl-2 protein, reduced positive apoptosis signals, and a decrease in the abundance of MKK 3/6 and p-p38 proteins (p < 0.05). In conclusion, dietary supplementation of PQQ could alleviate jejunal inflammatory damage and cell apoptosis in piglets challenged with LPS through the MKK3/6-p38 signaling pathway.

The Impact of Sea Buckthorn (Hippophae rhamnoides L.) and Cornelian Cherry (Cornus mas L.) Plant Extracts on the Physiology of Gastrointestinal Tract Cell In Vitro Model in the Context of Metabolic Diseases

The aim of this study was to evaluate the impact of ethanol extracts from sea buckthorn and Cornelian cherry fruits and leaves on physiology of gastrointestinal tract cells. We used three cell lines relevant to the types of cells, which are exposed to bioactive compounds after oral administration, namely intestinal absorptive cells (Caco-2/HT-29 MTX model), hepatocytes (HepG2 cells) and immunocompetent cells (RAW 264.7 and P388D1 monocytes). The contents of antioxidant and bioactive polyphenols, such as cinnamic, caffeic and p-coumaric acids, rutin, myricetin, resveratrol, quercetin, apigenin and kaempferol, were assessed in the extracts using HPLC chromatography. The application of the extracts to Caco-2/HT-29-MTX cultures increased enterocyte differentiation markers (alkaline phosphatase and villin1 level) and goblet cell markers (mucins) over a fortnight. The extracts reduced lipid droplet size in hepatocytes challenged with hyperglycaemic glucose concentration, insulin and palmitate. Sea buckthorn leaf, fruit and Cornelian cherry leaf extracts blocked oxidative burst in the PMA-stimulated monocytes, while the sea buckthorn leaf and Cornelian cherry fruit extracts downregulated lipopolysaccharide-induced NO and IL-1β, respectively. The results indicate that the tested extracts modulate the behaviour of cells in the gastrointestinal tract in a beneficial way, especially regarding lipid accumulation and innate immunity actions.

Effect of ultrasound treatment on quality parameters and health promoting activity of fish protein hydrolysates extracted from side streams of Atlantic mackerel (Scomber scombrus).

Fish protein hydrolysates (FPH) obtained by enzymatic hydrolysis allows for smart valorization of fish side streams. However, further treatments are normally needed to enhance bioactive and functional properties of the obtained FPH. At present, the commonly used methods to improve functional properties of FPH include chemical and enzymatic modification. Chemical treatments often cause environmental problems, while the enzymatic modification method requires the use of quite expensive enzymes. In recent years, emerging technologies such as ultrasound treatment (US-treatment) have shown great potential in protein modification with high efficiency and safety, low energy consumption, and low nutritional destructiveness. In this study, high-power ultrasound treatments were applied to fish protein hydrolysates (FPH) extracted from Atlantic mackerel (Scomber scombrus) side streams to improve their quality parameters. The effect of three different treatments of 300 W, 450 W and 600 W at the operating frequency of 20 kHz for 10 min on the physicochemical, structural, and functional characteristics of FPH, were examined. The results have shown that with an increase in ultrasound power, the protein solubility of FPH increased linearly, and the changes were significant for all US-treated samples compared to control (untreated) samples. US-treatment significantly increased the degree of hydrolysis of FPH samples treated with 450 W and 600 W compared to control samples. The carbonyl content of FPH increased (significantly for 450 W and 600 W), while thiol groups decreased (significantly for 300 W and 450 W). This indicated that some US-treatments induced oxidation of FPH, however the values of the protein oxidation were low. Amino acid composition of FPH revealed that US-treatment increased the proportion of essential amino acids in the sample treated with 300 W and 450 W, but the increase was not significant. After the US-treatment, all FPH samples became lighter and less yellowish and reddish, which suggest potentially higher attractiveness to consumers. In addition, the in vitro antioxidant activity was assessed using the DPPH, FRAP, and ABTS assays and the cell-free dipeptidyl peptidase IV (DPP-IV) inhibitory activity was also measured. Moreover, these biological activities were measured at cellular level utilizing human intestinal Caco-2 cells. Specifically, the FPH capacity to lower H2O2-induced reactive oxygen species (ROS) and lipid peroxidation levels was used to measure its antioxidant activity. The findings suggest that Scomber scombrus hydrolysates could find use as ingredients for promoting health.

Inflammation-induced epigenetic imprinting regulates intestinal stem cells

It remains unknown whether and how intestinal stem cells (ISCs) adapt to inflammatory exposure and whether the adaptation leaves scars that will affect their subsequent regeneration. We investigated the consequences of inflammation on Lgr5+ ISCs in well-defined clinically relevant models of acute gastrointestinal graft-versus-host disease (GI GVHD). Utilizing single-cell transcriptomics, as well as organoid, metabolic, epigenomic, and invivo models, we found that Lgr5+ ISCs undergo metabolic changes that lead to the accumulation of succinate, which reprograms their epigenome. These changes reduced the ability of ISCs to differentiate and regenerate exvivo in serial organoid cultures and also invivo following serial transplantation. Furthermore, ISCs demonstrated a reduced capacity for invivo regeneration despite resolution of the initial inflammatory exposure, demonstrating the persistence of the maladaptive impact induced by the inflammatory encounter. Thus, inflammation imprints the epigenome of ISCs in a manner that persists and affects their sensitivity to adapt to future stress or challenges.