Journal: iScience

Article Title: LRMDA promotes intestinal mucosal barrier integrity and limits colitis and colon cancer

doi: 10.1016/j.isci.2026.116785

Figure Lengend Snippet: LRMDA deficiency impairs colonic mucosal barrier by reducing mucin secretion (A) Representative images of PAS/AB-stained distal colon sections of Lrmda +/+ ( n = 19) and Lrmda −/− ( n = 17) mice. (B) Quantification of inner mucus layer thickness in the distal colons of mice as in (A). (C) Representative scanning election microscopy images of the distal colon of Lrmda +/+ ( n = 6) and Lrmda −/− ( n = 6) mice. (D) Area (left) and number (right) of mucin granules in the distal colon of Lrmda +/+ and Lrmda −/− mice from (C). (E) Transmission electron microscopy images of goblet cells of Lrmda +/+ ( n = 6) and Lrmda −/− ( n = 6) mice. (F) A scheme of FITC-dextran treatment model (left) and concentration of FITC-dextran in serum (right) of Lrmda +/+ ( n = 6) and Lrmda −/− ( n = 6) at the 4th hour after being deprived of food overnight and being injected with FITC-dextran (500 mg per kg body weight, i.g.) by gavage. (G) Representative immunofluorescence imaging (anti-FITC) analysis of the distal colon sections of Lrmda +/+ and Lrmda −/− mice treated as in (F). (H) FISH (EUB338) combined with immunofluorescence imaging (anti-MUC2) analysis of the distal colon sections of Lrmda +/+ ( n = 6) and Lrmda −/− ( n = 6) mice. ns, no significance; ∗ p < 0.05; ∗∗∗∗ p < 0.0001. Two-tailed Student’s t test in (B), (D), and (F). Mean ± SEM (B), (D), and (F). Scale bars represent 100 μm in (A), (C), and (G); 5 μm in (E); and 50 μm in (H), respectively. Data are representative results of three independent experiments (A–D, F, and G) or two independent experiments (E and H).

Article Snippet: Anti-rabbit MUC2 , Abclonal , Cat# A4767.

Techniques: Staining, Microscopy, Transmission Assay, Electron Microscopy, Concentration Assay, Injection, Immunofluorescence, Imaging, Two Tailed Test

Journal: Microbiome

Article Title: Gut microbiota-derived riboflavin mediates protective effects of MUC2 in acute pancreatitis via CD40-macrophage pathway

doi: 10.1186/s40168-026-02375-z

Figure Lengend Snippet: Intestinal Muc2 knockout aggravated pancreatic injuries in AP. A PAS-staining and the numbers of mucus-containing goblet cells in the duodenum of AP patients with different etiologies. Original magnification, ×200. B Immunofluorescence and quantification of MUC2 in the duodenum of AP patients with different etiologies. Original magnification, ×200. C Histopathological changes and pathological scores of the pancreas. Original magnification, ×100 (top panels) or ×200 (bottom panels). D Serum amylase activity. E TUNEL staining and quantification of TUNEL + cells in pancreas. Original magnification, ×200. F Real-time PCR analysis of IL-1β, TNF-α, IL-6 mRNA levels in pancreas; tubulin was used as the reference gene. G ELISA analysis of serum IL-1β, TNF-α, and IL-6 levels. The data are presented as the means ± SEM ( n = 6 mice per group). ns, no significant difference; * p < 0.05; HC, healthy controls; ABP, acute biliary pancreatitis; HTGAP, hypertriglyceridemic acute pancreatitis; AP, acute pancreatitis (cerulein); WT, Muc2 flox/flox littermate control; Muc2 ΔIEC , mice with specific deletion of Muc2 in intestinal epithelium

Article Snippet: Membranes were blocked with 3% bovine serum albumin (BSA) for 1 h, then incubated overnight at 4 °C with primary antibodies against MUC2 (A14659; Abclonal, China) or CD40 (A22714; Abclonal, China) in primary antibody dilution buffer (Epizyme Biotech, China).

Techniques: Knock-Out, Staining, Immunofluorescence, Activity Assay, TUNEL Assay, Real-time Polymerase Chain Reaction, Analysis, Enzyme-linked Immunosorbent Assay, Control

Journal: Microbiome

Article Title: Gut microbiota-derived riboflavin mediates protective effects of MUC2 in acute pancreatitis via CD40-macrophage pathway

doi: 10.1186/s40168-026-02375-z

Figure Lengend Snippet: Intestinal Muc2 knockout aggravated AP-related intestinal injuries. A Histopathological changes and pathological scores of the colon. Original magnification, ×100 (top panels) or ×200 (bottom panels). B TUNEL staining and quantification of TUNEL + cell in the colon. Original magnification, ×200. C Immunofluorescence staining for Claudin-1 and Occludin (green) in the colon. Original magnification, ×100. D Real-time PCR analysis of IL-1β, TNF-α, IL-6 mRNA levels in the colon; tubulin was used as the reference gene. E ELISA analysis of serum DAO, D-lactate levels. F FISH detection of bacteria translocation in the pancreas and colon by the EUB338 probe. Original magnification, ×200. G Colony-forming units (CFUs) of anaerobic bacteria in mesenteric lymph nodes and liver. The data are presented as the means ± SD ( n = 6 mice per group). ns, no significant difference; * p < 0.05; WT, Muc2 flox/flox littermate control; Muc2 ΔIEC , mice with specific deletion of Muc2 in intestinal epithelial cells; AP, acute pancreatitis. FISH, fluorescence in situ hybridization

Article Snippet: Membranes were blocked with 3% bovine serum albumin (BSA) for 1 h, then incubated overnight at 4 °C with primary antibodies against MUC2 (A14659; Abclonal, China) or CD40 (A22714; Abclonal, China) in primary antibody dilution buffer (Epizyme Biotech, China).

Techniques: Knock-Out, TUNEL Assay, Staining, Immunofluorescence, Real-time Polymerase Chain Reaction, Analysis, Enzyme-linked Immunosorbent Assay, Bacteria, Translocation Assay, Control, Fluorescence, In Situ Hybridization

Journal: Microbiome

Article Title: Gut microbiota-derived riboflavin mediates protective effects of MUC2 in acute pancreatitis via CD40-macrophage pathway

doi: 10.1186/s40168-026-02375-z

Figure Lengend Snippet: Muc2 deficiency aggravated intestinal microbiota dysbiosis and reduced riboflavin production. A The bar graph of the relative abundance of the top 20 genera in feces between WT and Muc2 ΔIEC groups. B The bar graph of the relative abundance of the top 20 genera in feces between AP and Muc2 ΔIEC + AP groups. C KEGG enrichment analysis of differential fecal metabolites between WT and Muc2 ΔIEC groups. D KEGG enrichment analysis of differential fecal metabolites between AP and Muc2 ΔIEC + AP groups. E The VIP value plots of the differential fecal metabolites (the top 20 of VIP value) between WT and Muc2 ΔIEC groups. F The VIP value plots of the differential fecal metabolites (the top 20 of VIP value) between AP and Muc2 ΔIEC + AP groups. G Riboflavin, flavin mononucleotide, and flavin adenine dinucleotide levels in feces. H Targeted metabolomics analysis of riboflavin, flavin mononucleotide and flavin adenine dinucleotide levels in pancreas. I Network analysis between gut microbiome and fecal metabolites. Data are presented as the means ± SEM ( n = 5 or 6 mice per group). ns, no significant difference; * p < 0.05; WT, Muc2 flox/flox littermate control; Muc2 ΔIEC , mice with specific deletion of Muc2 in intestinal epithelium; AP, acute pancreatitis

Article Snippet: Membranes were blocked with 3% bovine serum albumin (BSA) for 1 h, then incubated overnight at 4 °C with primary antibodies against MUC2 (A14659; Abclonal, China) or CD40 (A22714; Abclonal, China) in primary antibody dilution buffer (Epizyme Biotech, China).

Techniques: Analysis, Microbiome, Control

Journal: Microbiome

Article Title: Gut microbiota-derived riboflavin mediates protective effects of MUC2 in acute pancreatitis via CD40-macrophage pathway

doi: 10.1186/s40168-026-02375-z

Figure Lengend Snippet: Rib deficiency weakened the protective effect of Lactobacillus plantarum on AP in Muc2 ΔIEC mice. A Schematic diagram of L. plantarum AR113 with Rib deletion (AR113Δ Rib ). B Experiment design and time axis of AR113Δ Rib gavage and AP induction in Muc2 ΔIEC mice. C Histopathological changes and pathological scores of the pancreas. Original magnification, ×100 (top panels) or ×200 (bottom panels). D Histopathological changes and pathological scores of the colon. Original magnification, ×100 (top panels) or ×200 (bottom panels). E ELISA analysis of serum IL-1β, TNF-α, and IL-6 levels. F Real-time PCR analysis of IL-1β, TNF-α, and IL-6 mRNA levels in the colon. Tubulin was used as the reference gene. G Immunofluorescence staining for claudin1and occludin (green) in the colon. Original magnification, ×200. H FISH detection of bacteria translocation in the pancreas by the EUB338 probe. Original magnification, ×200. The data are presented as the means ± SEM ( n = 6 mice per group). * p < 0.05; Muc2 ΔIEC , mice with specific deletion of Muc2 in intestinal epithelium; AR113, L. plantarum AR113; AR113Δ Rib , L. plantarum AR113 with Rib deletion; AP, acute pancreatitis; FISH, fluorescence in situ hybridization

Article Snippet: Membranes were blocked with 3% bovine serum albumin (BSA) for 1 h, then incubated overnight at 4 °C with primary antibodies against MUC2 (A14659; Abclonal, China) or CD40 (A22714; Abclonal, China) in primary antibody dilution buffer (Epizyme Biotech, China).

Techniques: Enzyme-linked Immunosorbent Assay, Analysis, Real-time Polymerase Chain Reaction, Immunofluorescence, Staining, Bacteria, Translocation Assay, Fluorescence, In Situ Hybridization

Journal: Microbiome

Article Title: Gut microbiota-derived riboflavin mediates protective effects of MUC2 in acute pancreatitis via CD40-macrophage pathway

doi: 10.1186/s40168-026-02375-z

Figure Lengend Snippet: Riboflavin alleviated AP in Muc2 ΔIEC mice. A Schematic diagram of riboflavin treatment. B Histopathological changes and pathological scores of the pancreas. Original magnification, ×100 (top panels) or ×200 (bottom panels). C Serum amylase activity. D ELISA analysis of serum IL-1β, TNF-α, and IL-6 levels. E Histopathological changes and pathological scores of the colon. Original magnification, ×100 (top panels) or ×200 (bottom panels). F Real-time PCR analysis of IL-1β, TNF-α, and IL-6 mRNA levels in the colon. Tubulin was used as the reference gene. G Immunofluorescence staining for claudin-1and occludin (green) in the colon. Original magnification, ×200. H FISH detection of bacteria translocation in the pancreas and colon by the EUB338. I CFUs of anaerobic bacteria in liver and mesenteric lymph nodes. The data are presented as the means ± SEM ( n = 6 mice per group). * p < 0.05; Muc2 ΔIEC , mice with specific deletion of Muc2 in intestinal epithelium; AP, acute pancreatitis; RF, riboflavin; CFUs, colony-forming units

Article Snippet: Membranes were blocked with 3% bovine serum albumin (BSA) for 1 h, then incubated overnight at 4 °C with primary antibodies against MUC2 (A14659; Abclonal, China) or CD40 (A22714; Abclonal, China) in primary antibody dilution buffer (Epizyme Biotech, China).

Techniques: Activity Assay, Enzyme-linked Immunosorbent Assay, Analysis, Real-time Polymerase Chain Reaction, Immunofluorescence, Staining, Bacteria, Translocation Assay

Journal: Microbiome

Article Title: Gut microbiota-derived riboflavin mediates protective effects of MUC2 in acute pancreatitis via CD40-macrophage pathway

doi: 10.1186/s40168-026-02375-z

Figure Lengend Snippet: Riboflavin reduced the pro-inflammatory macrophage infiltration in AP mice with intestinal Muc2 knockout. A Immunofluorescence staining and quantification for pro-inflammatory macrophages (F4/80 + NOS2 + ) in the pancreas. Original magnification, ×200. B Immunofluorescence staining and quantification for neutrophil (Ly6G + ). Original magnification, ×200. C Flow cytometry analysis and quantification of pro-inflammatory macrophages (F4/80 + CD86 + ) and anti-inflammatory macrophages (F4/80 + CD206 + ) in the pancreas. D Flow cytometry analysis and quantification of neutrophil (CD11b + Ly6G +) . E Immunofluorescence staining and quantification for Th17 cells (CD4 + RORγ + ). Original magnification, ×200. F Immunofluorescence staining and quantification for Treg cells (CD4 + FOXP3 + ). Original magnification, ×200. G Immunofluorescence staining and quantification for pro-inflammatory macrophages (F4/80 + NOS2 + ) in the pancreas. Original magnification, ×200. H Real-time PCR analysis of Nos2 mRNA levels in macrophage with different concentrations of riboflavin. Tubulin was used as the reference gene. I Real-time PCR analysis of Nos2, TNF-α, IL-6, and IL-1β mRNA levels. Tubulin was used as the reference gene. J Immunofluorescence staining for NOS2 (red). Original magnification, ×200. The data are presented as the means ± SEM ( n = 6 mice per group). ns, no significant difference; * p < 0.05; WT, Muc2 flox/flox littermate control; Muc2 ΔIEC , mice with specific deletion of Muc2 in intestinal epithelium; AP, acute pancreatitis; RF, riboflavin

Article Snippet: Membranes were blocked with 3% bovine serum albumin (BSA) for 1 h, then incubated overnight at 4 °C with primary antibodies against MUC2 (A14659; Abclonal, China) or CD40 (A22714; Abclonal, China) in primary antibody dilution buffer (Epizyme Biotech, China).

Techniques: Knock-Out, Immunofluorescence, Staining, Flow Cytometry, Analysis, Real-time Polymerase Chain Reaction, Control

Journal: Microbiome

Article Title: Gut microbiota-derived riboflavin mediates protective effects of MUC2 in acute pancreatitis via CD40-macrophage pathway

doi: 10.1186/s40168-026-02375-z

Figure Lengend Snippet: Reduced intestinal MUC2 was associated with decreased serum riboflavin in AP patients. A Spearman correlation analysis of intestinal MUC2 expression level and serum riboflavin concentration in the initial patient cohort. B Analysis of intestinal MUC2 expression levels between MAP/MSAP and SAP. C Analysis of serum riboflavin concentrations between MAP/MSAP and SAP. D Spearman correlation analysis of intestinal MUC2 expression level and Ranson score in the initial patient cohort. E Spearman correlation analysis of serum riboflavin concentration and Ranson score in the initial patient cohort. F Spearman correlation analysis of serum riboflavin concentration and Ranson score in the validation cohort. AP, acute pancreatitis; MAP, mild acute pancreatitis; MSAP, moderately severe acute pancreatitis; SAP, severe acute pancreatitis

Article Snippet: Membranes were blocked with 3% bovine serum albumin (BSA) for 1 h, then incubated overnight at 4 °C with primary antibodies against MUC2 (A14659; Abclonal, China) or CD40 (A22714; Abclonal, China) in primary antibody dilution buffer (Epizyme Biotech, China).

Techniques: Analysis, Expressing, Concentration Assay, Biomarker Discovery

Journal: Research

Article Title: AI-Powered Discovery of Rosmarinic Acid as a Novel Ferroptosis Inhibitor for Ulcerative Colitis via Targeting the ALOX15–VDAC1 Axis

doi: 10.34133/research.1352

Figure Lengend Snippet: Attenuation of DSS-induced colitis by rosmarinic acid through suppression of ALOX15-dependent ferroptosis and preservation of intestinal barrier function. (A) The daily weight change of each group of mice. (B) Disease activity index (DAI) score of each group of colitis mice. (C) Representative images of colons from each group. (D) Representative H&E and immunohistochemical staining images of colon tissues showing the expression of intestinal barrier proteins in each group. Image captured at ×20 magnification; scale bar: 100 μm. (E) Quantitative analysis of the expression levels of intestinal barrier proteins (ZO-1, Occludin, Claudin-1, and MUC-2) and ALOX15 using ImageJ software based on previously shown IHC staining images. (F) Relative protein expression levels of ferroptosis-related markers (ACSL4, COX-2, ALOX15, GPX4, FTH1, and SLC7A11) in mouse colon tissue assessed by Western blotting and quantified using ImageJ software. (G) Relative protein expression levels and quantitative analysis of tight junction proteins (ZO-1, Occludin, and Claudin-1). Compared with the DSS group: * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001.

Article Snippet: Primary antibodies targeting ZO-1, Occludin, Claudin-1, MUC-2, and ALOX15 (ABclonal, China), diluted in 5% bovine serum albumin (BSA), were then applied overnight at 4 °C.

Techniques: Preserving, Activity Assay, Immunohistochemical staining, Staining, Expressing, Software, Immunohistochemistry, Western Blot