Journal: Advanced Science

Article Title: Gut Microbiota‐Derived Indole‐3‐Acetaldehyde Contributes to the Disease‐Alleviating Effects of Kurarinone in Experimental Autoimmune Encephalomyelitis

doi: 10.1002/advs.78358

Figure Lengend Snippet: KU treatment mitigates EAE and reduces Th1 and Th17 cell proportions in mice. (A) Schematic timeline of EAE induction and KU administration (100 mg kg −1 , intragastric). Mice were immunized with MOG 35‐55 peptide, and KU or vehicle was administered from 11 to 21 dpi. (B) Line graph of clinical scores in the control (Ctrl), vehicle‐treated EAE, and KU‐treated EAE groups ( n = 8 per group). (C) Disease incidence curves of EAE mice. (D) Cumulative clinical scores and peak score in each group. (E) Representative HE staining and LFB staining of spinal cord from the Ctrl, vehicle‐treated EAE, and KU‐treated EAE groups. (F) Inflammation and demyelination scores based on HE and LFB staining ( n = 6 per group). (G) The proportion of CD3 + CD4 + T cells in the CNS and spleen (SP) ( n = 3–4 per group). (H) Absolute counts of CD3 + and CD4 + T cells in the CNS of each group. (I) The proportion of CD4 + IFNγ + (Th1) cells in the CNS and SP. (J) The proportion of CD4 + IL‐17A + (Th17) cells in the CNS and SP. (K) Relative mRNA expression of T‐bet and RORγt in the spinal cord measured by qPCR. (L) ELISA detection of serum IFNγ and IL‐17A concentrations in each group. Data are presented as mean ± SEM; * p < 0.05, ** p < 0.01, *** p < 0.001 vs. vehicle‐treated EAE.

Article Snippet: The mouse serum was detected using the IFNγ and IL‐17A ELISA kits (Elabscience, E‐EL‐M0047, E‐EL‐M0048) according to the kit instructions.

Techniques: Control, H&E Stain, Staining, Expressing, Enzyme-linked Immunosorbent Assay

Journal: Advanced Science

Article Title: Gut Microbiota‐Derived Indole‐3‐Acetaldehyde Contributes to the Disease‐Alleviating Effects of Kurarinone in Experimental Autoimmune Encephalomyelitis

doi: 10.1002/advs.78358

Figure Lengend Snippet: KU modulates gut microbiota to ameliorate EAE. (A) Alpha diversity (Shannon index) of gut microbiota in the Ctrl, vehicle‐treated EAE, and KU‐treated EAE groups ( n = 8 per group). Pair‐wise comparisons were performed using Wilcoxon rank‐sum test. (B) Principal coordinate analysis (PCoA) of gut microbiota based on Bray‐Curtis distance (pseudo‐ F = 30.12, R 2 = 0.741, p < 0.001); each dot represents an individual sample. (C) Stacked bar plot showing the relative abundance of gut microbiota at the family level in the Ctrl, vehicle‐treated EAE, and KU‐treated EAE groups. (D) Schematic timeline of ABX treatment and FMT and EAE induction: EAE mice received FMT from vehicle‐treated or KU‐treated donor mice (100 mg kg −1 , intragastric). (E) Clinical scores of FMT‐Vehicle and FMT‐KU groups ( n = 9 per group). (F) Disease incidence curves of EAE mice. (G) Statistical analysis of cumulative clinical scores and peak score. (H) Representative HE staining and LFB staining of spinal cord tissues from FMT‐Vehicle and FMT‐KU mice. (I) Inflammation score and demyelination score in the spinal cord ( n = 4 per group). (J) Flow cytometric plots of CD4 + IFNγ + (Th1) and CD4 + IL‐17A + (Th17) cells in the CNS and SP. (K) Quantification of CD4 + IFNγ + and CD4 + IL‐17A + cell percentages in the CNS and SP ( n = 5 per group). Data are presented as mean ± SEM. * p < 0.05, ** p < 0.01, *** p < 0.001 vs. EAE mice that received FMT from vehicle‐treated donors.

Article Snippet: The mouse serum was detected using the IFNγ and IL‐17A ELISA kits (Elabscience, E‐EL‐M0047, E‐EL‐M0048) according to the kit instructions.

Techniques: Analysis, H&E Stain, Staining

Journal: Advanced Science

Article Title: Gut Microbiota‐Derived Indole‐3‐Acetaldehyde Contributes to the Disease‐Alleviating Effects of Kurarinone in Experimental Autoimmune Encephalomyelitis

doi: 10.1002/advs.78358

Figure Lengend Snippet: KU ameliorates EAE in an A. muciniphila ‐dependent manner. (A) Heatmap showing the relative abundance of the top 10 gut microbial species in the Ctrl, vehicle‐treated EAE, and KU‐treated groups ( n = 8 per group). (B) Linear discriminant analysis effect size (LEfSe) analysis was performed to screen differentially abundant microbial taxa across the three groups. Taxa with an LDA score > 3 were defined as potential microbial biomarkers with statistical significance. (C) Schematic timeline of ABX pretreatment, KU administration, A. muciniphila supplementation, and EAE induction. (D) Clinical scores of EAE, EAE+ABX, EAE+ABX+KU, and EAE+ABX+KU+ A. muciniphila ( n = 8 per group). (E) Cumulative clinical scores and peak score in each group. (F) Representative HE staining and LFB staining of the spinal cord. (G) Spinal cord inflammation score and demyelination score ( n = 4 per group). (H) Flow cytometric plots of CD4 + IFNγ + (Th1) cells in the CNS and SP of each group. (I) Quantification of CD4 + IFNγ + cell percentages in the CNS and SP ( n = 4 per group). (J) Flow cytometric plots of CD4 + IL‐17A + (Th17) cells in the CNS and SP of each group. (K) Quantification of CD4 + IL‐17A + cell percentages in the CNS and SP ( n = 4 per group). Data are presented as mean ± SEM. * p < 0.05, ** p < 0.01, *** p < 0.001; ns, p > 0.05.

Article Snippet: The mouse serum was detected using the IFNγ and IL‐17A ELISA kits (Elabscience, E‐EL‐M0047, E‐EL‐M0048) according to the kit instructions.

Techniques: Analysis, H&E Stain, Staining

Journal: Advanced Science

Article Title: Gut Microbiota‐Derived Indole‐3‐Acetaldehyde Contributes to the Disease‐Alleviating Effects of Kurarinone in Experimental Autoimmune Encephalomyelitis

doi: 10.1002/advs.78358

Figure Lengend Snippet: IAAld alleviates EAE and reduces Th1 and Th17 cell proportions. (A) Schematic timeline of EAE induction and IAAld administration ( n = 8 per group). (B) Clinical scores of vehicle‐treated EAE, EAE+IAAld 40 mg/kg/day (IAAld 40), and EAE+IAAld 80 mg/kg/day (IAAld 80) groups. (C) Disease incidence curves of EAE mice with different doses of IAAld treatment. (D) Quantification of cumulative clinical scores and peak score in each group. (E) Representative HE staining and LFB staining of spinal cord tissues from the vehicle‐treated EAE, IAAld 40, and IAAld 80 groups. (F) Spinal cord inflammation score and demyelination score ( n = 3 per group). (G) Flow cytometric plots of CD4 + IFNγ + (Th1) and CD4 + IL‐17A + (Th17) cells in the CNS and SP of each group. (H) Quantification of CD4 + IFNγ + and CD4 + IL‐17A + cell percentages in the CNS and SP ( n = 4 per group). Data are presented as mean ± SEM. * p < 0.05, ** p < 0.01, *** p < 0.001.

Article Snippet: The mouse serum was detected using the IFNγ and IL‐17A ELISA kits (Elabscience, E‐EL‐M0047, E‐EL‐M0048) according to the kit instructions.

Techniques: H&E Stain, Staining

Journal: Advanced Science

Article Title: Gut Microbiota‐Derived Indole‐3‐Acetaldehyde Contributes to the Disease‐Alleviating Effects of Kurarinone in Experimental Autoimmune Encephalomyelitis

doi: 10.1002/advs.78358

Figure Lengend Snippet: IAAld suppresses Th1 and Th17 cell differentiation in an AhR‐dependent manner by repressing JAK/STAT signaling. (A) KEGG pathway enrichment analysis of the intersectional targets of IAAld and MS. (B) In silico prediction of the binding interaction between IAAld and AhR by molecular docking. The calculated binding energy (BE) is −6.6 kcal/mol (inset: chemical structure of IAAld). (C,D) Flow cytometric quantification and representative plots of CD4 + IFNγ + (Th1) and CD4 + IL‑17A + (Th17) cells under in vitro differentiation conditions across PBS, IAAld (10 µM), and IAAld+siAhR (AhR knockdown by siRNA) groups ( n = 3 per group). (E) Immunofluorescence staining of AhR (green) in Th1 and Th17 cells of each group, and cell nuclei were stained with DAPI (blue). (F) Western blot analysis showing protein levels of AhR, p‑JAK2, JAK2, p‑STAT4, STAT4, p‑STAT3, STAT3, SOCS1, SOCS3, and GAPDH in Th1 (left) and Th17 (right) cells. (G) Quantitative analysis of AhR/GAPDH ratio, p‐JAK2/JAK2 ratio, p‐STAT4/STAT4 ratio, p‐STAT3/STAT3 ratio, SOCS1/GAPDH ratio and SOCS3/GAPDH ratio in each group ( n = 4 per group). Data are presented as mean ± SEM. * p < 0.05, ** p < 0.01, *** p < 0.001.

Article Snippet: The mouse serum was detected using the IFNγ and IL‐17A ELISA kits (Elabscience, E‐EL‐M0047, E‐EL‐M0048) according to the kit instructions.

Techniques: Cell Differentiation, Analysis, In Silico, Binding Assay, Docking Assay, In Vitro, Knockdown, Immunofluorescence, Staining, Western Blot, Significance Assay

Journal: Advanced Science

Article Title: Gut Microbiota‐Derived Indole‐3‐Acetaldehyde Contributes to the Disease‐Alleviating Effects of Kurarinone in Experimental Autoimmune Encephalomyelitis

doi: 10.1002/advs.78358

Figure Lengend Snippet: IAAld treatment mitigates EAE in an AhR‐dependent manner. (A) Relative mRNA expression levels of Ahr, Cyp1a1, and Cyp1b1 in the spinal cord of EAE mice treated with vehicle, IAAld 40, or IAAld 80 groups. (B) Schematic timeline depicting the experimental protocol for EAE induction, followed by administration of IAAld (80 mg/kg/day) and AhR inhibitor (AhRi; SR1, 50 mg/kg/day). (C) Clinical scores for EAE mice treated with vehicle, IAAld, and IAAld+AhRi groups ( n = 6 per group). (D) Disease incidence curves for EAE mice over time. (E) Quantification of cumulative clinical scores and peak score for each group. (F) Representative HE and LFB staining of spinal cord tissues for assessment of inflammation and demyelination, respectively. (G) Quantification of spinal cord inflammation and demyelination scores ( n = 3 per group). (H) Representative flow cytometry plots indicating the percentages of CD4 + IFNγ + (Th1) and CD4 + IL‐17A + (Th17) cells in the CNS and SP of each group. (I) Quantification of CD4 + IFNγ + and CD4 + IL‐17A + cell percentages in the CNS and SP across groups ( n = 3–4 per group). Data are presented as mean ± SEM. * p < 0.05, ** p < 0.01, *** p < 0.001.

Article Snippet: The mouse serum was detected using the IFNγ and IL‐17A ELISA kits (Elabscience, E‐EL‐M0047, E‐EL‐M0048) according to the kit instructions.

Techniques: Expressing, Staining, Flow Cytometry