Journal: bioRxiv

Article Title: Enteric ChAT-expressing neurons as new target for Lactobacillus plantarum ameliorates inflammatory bowel diseases

doi: 10.1101/2025.09.09.673902

Figure Lengend Snippet: (A) Schematic illustration from human subjects. Ulcerative colitis patients (UC, n=35), healthy controls (HC, n=26). (B) Representative endoscopic images. (C) Levels of hemoglobin in the blood. (D-F) Leukocyte (D), monocyte (E) and neutrophil (F) counts in the blood. (G) Principal co-ordinates analysis (PCoA) of unweighted unifrac distance based on the 16S rRNA sequencing from stool samples. ANOSIM (analysis of similarities), R = 0.1, p = 0.007. (H) Linear discriminative analysis (LDA) score of the top 15 most differential genera from HC and UC. (I) Relative abundance of Lactobacillus genus from HC and UC. (J-M) Correlative analysis of Lactobacillus abundance with C-reactive protein (CRP) values (J), leukocyte (K), monocyte (L) and neutrophil (M) counts in UC patients. (N)Venn diagram indicating genus enriched in the stool samples from healthy people and normal mice (LDA score > 3 and p < 0.05 using Wilcoxon rank-sum test). Mean ± SEM shown. ns, not significant, *p < 0.05, **p < 0.01, ***p < 0.001, and ****p < 0.0001, determined by unpaired Student’s t-test. See also Figure S1, Table S1 and S2.

Article Snippet: For antibacterial activity of the Lactobacillus strains against pathogen bacteria such as Staphylococcus aureus ATCC 6538 and Escherichia coli ATCC 700928, the Oxford cup method was performed according to previous study., Petri plates were poured with sterile Luria Bertani (LB) agar medium and seeded with 200 μL freshly grown pathogen bacteria, followed by wells of 9 mm diameter excavated with a sterilized Oxford cup.

Techniques: Sequencing

Journal: bioRxiv

Article Title: Enteric ChAT-expressing neurons as new target for Lactobacillus plantarum ameliorates inflammatory bowel diseases

doi: 10.1101/2025.09.09.673902

Figure Lengend Snippet: (A) The abundance of Lactobacillus genus from UC patients in diverse disease course by a previous report. (B) Body weight change. (C) Disease activity index (DAI). (D) Representative image of hematoxylin and eosin (H&E)-staining sections in colon. Scale bar, 50 μm. (E) Histological scores. (F) Principal co-ordinates analysis (PCoA) of stool samples based on unweighted unifrac distances by 16S rRNA sequencing. ANOSIM (analysis of similarities), R = 0.983259, p = 0.001. (G) Averaged relative abundance of bacteria at the genus level from Control and DSS. (H) Linear discriminative analysis (LDA) score of differentially enriched bacterial genera measured by Linear discriminant analysis Effect Size (LEfSe) analysis between Control and DSS. (I) Relative abundance of Lactobacillus genus from Control and DSS in mice. (J-K) Correlation analysis of Lactobacillus abundance with DAI (J) and histological scores (K). Control group (Control, n=8) DSS group (DSS, n=8). Mean ± SEM shown. ns, not significant, *p < 0.05, **p < 0.01, ***p < 0.001, and ****p < 0.0001, determined by unpaired Student’s t-test.

Article Snippet: For antibacterial activity of the Lactobacillus strains against pathogen bacteria such as Staphylococcus aureus ATCC 6538 and Escherichia coli ATCC 700928, the Oxford cup method was performed according to previous study., Petri plates were poured with sterile Luria Bertani (LB) agar medium and seeded with 200 μL freshly grown pathogen bacteria, followed by wells of 9 mm diameter excavated with a sterilized Oxford cup.

Techniques: Activity Assay, Staining, Sequencing, Bacteria, Control

Journal: bioRxiv

Article Title: Enteric ChAT-expressing neurons as new target for Lactobacillus plantarum ameliorates inflammatory bowel diseases

doi: 10.1101/2025.09.09.673902

Figure Lengend Snippet: (A-N) Growth curve of L. plantarum (A), L. paracasei (B), L.fermentum (C), L. alimentarius (D), L. brevis (E), L. delbrueckii (F), L.casei (G), L.helveticus (H), L.reuteri (I), L. nagelii (J), L. sakei (K), L.oryzae (L), L.pentosus (M), and L.coryn iformis (N) (n=3). (O) The survival rate of Lactobacillus strains following simulated gastrointestinal fluids. (P) Antibacterial activity. (Q) Diagram of a Caco-2 monolayer. (R) Adhesion of the Lactobacillus strains with Caco-2 cell monolayers (MOE 10, n=3). (S) 4 kDa FITC-dextran paracellular transport from the apical to basolateral compartment on a Caco-2 cell monolayer pretreated with Lactobacillus strains (MOE; 10, 24 h) before TNFα exposure (20 ng/mL, 24 h, n=3). (T) Loading plot of principal component analysis (PCA). (U) Score plot of PCA based on the survival rate, antibacterial activity, adhesion and intestinal permeability of Lactobacillus strains in vitro. (V) Fuzzy synthetic evaluation (FSE) analysis. Mean ± SEM shown. Data are determined by one-way ANOVA with Tukey’s multiple comparison test. ns, not significant, *p < 0.05, **p < 0.01, ***p < 0.001, and ****p < 0.0001, compared with the Control group, # p<0.05, ## p<0.01, ### p < 0.001, and #### p < 0.0001, compared with the TNFα group.

Article Snippet: For antibacterial activity of the Lactobacillus strains against pathogen bacteria such as Staphylococcus aureus ATCC 6538 and Escherichia coli ATCC 700928, the Oxford cup method was performed according to previous study., Petri plates were poured with sterile Luria Bertani (LB) agar medium and seeded with 200 μL freshly grown pathogen bacteria, followed by wells of 9 mm diameter excavated with a sterilized Oxford cup.

Techniques: Activity Assay, Permeability, In Vitro, Comparison, Control

Journal: bioRxiv

Article Title: Enteric ChAT-expressing neurons as new target for Lactobacillus plantarum ameliorates inflammatory bowel diseases

doi: 10.1101/2025.09.09.673902

Figure Lengend Snippet: (A) Experimental diagram for Lactobacillus strains administration schedule. (B) Body weight change (n=8). (C) Body weight change at day 8 after DSS induction (n=8). (D) Disease activity index (DAI) score (n=8). (E) DAI at day 8 after DSS induction (n=8). (F) Representative photographs of colon. (G) Colon length (n=8). (H and I) Representative image of hematoxylin and eosin (H&E)-staining sections in colon and histological scores (n=6). Scale bar, 50 μm. (J-K) Relative gene expression of proinflammatory cytokines including TNF-α (J) and IL-1β (K) in colonic tissues accessed by quantitative real-time PCR. (L-M) AB/PAS staining of the distal colon and elevation of goblet cells per crypt (n=3). Scale bar, 50 μm. (N-O) Representative immunostaining images and quantification of Muc2 in the colon (n=3). Scale bar, 50 μm. (P) Relative gene expression of Muc2 in colonic tissues accessed by quantitative real-time PCR. Data are determined by two-way ANOVA with Tukey’s multiple comparison test (B-E) or one-way ANOVA with Tukey’s multiple comparison test (G-O). ns, not significant, *p < 0.05, **p < 0.01, ***p < 0.001, and ****p < 0.0001, compared with the Control group, # p<0.05, ## p<0.01, ### p < 0.001, and #### p < 0.0001, compared with the DSS group.

Article Snippet: For antibacterial activity of the Lactobacillus strains against pathogen bacteria such as Staphylococcus aureus ATCC 6538 and Escherichia coli ATCC 700928, the Oxford cup method was performed according to previous study., Petri plates were poured with sterile Luria Bertani (LB) agar medium and seeded with 200 μL freshly grown pathogen bacteria, followed by wells of 9 mm diameter excavated with a sterilized Oxford cup.

Techniques: Activity Assay, Staining, Gene Expression, Real-time Polymerase Chain Reaction, Immunostaining, Comparison, Control