Journal: Frontiers in Cellular and Infection Microbiology

Article Title: Curcumin β-D-Glucuronide Modulates an Autoimmune Model of Multiple Sclerosis with Altered Gut Microbiota in the Ileum and Feces

doi: 10.3389/fcimb.2021.772962

Figure Lengend Snippet: Effects of curcumin monoglucuronide (CMG) in an autoimmune model of multiple sclerosis (MS), experimental autoimmune encephalomyelitis (EAE). (A) For EAE induction, we sensitized C57BL/6 mice with the myelin oligodendrocyte glycoprotein (MOG) 35-55 peptides. We divided the mice into four groups: Control, Induction, Latent, and Whole groups (n = 7–9 per group), and treated the groups with CMG on days 0–4 (Induction), on days 11–15 (Latent), or throughout the course (Whole). We treated the control mice (Control) with phosphate-buffered saline (PBS). All CMG-treated groups had less severe EAE compared with the control group. The Whole group tended to have a lower EAE score than the Control group 30 days post-induction (p.i.) ( P = 0.06, Mann–Whitney U test). The Latent group tended to recover from acute EAE more quickly than the Control group 35 days p.i. ( P = 0.056, Mann–Whitney U test). The clinical score was calculated as the mean ± standard error (SE) of seven to nine mice per group. (B) We monitored body weight changes of mice daily. The levels of body weight changes had an inverse relationship to the severity of clinical signs. The Control EAE group had more weight loss compared with the Induction ( P = 0.06), Latent ( P = 0.07), and Whole ( P = 0.14) groups on day 34. Results are the mean ± SE of seven to nine mice per group. (C–F) We stained the spinal cord sections (scale bar: C and D = 500 µm, E and F = 50 µm) from EAE mice with Luxol fast blue (C, D) or anti-CD3 antibody (E, F) to visualize myelin or T cells, respectively, compared the spinal cord pathology between the Control (C, E) and CMG-treated Whole groups (D, F) . The Whole group had less severe demyelination (arrowhead), meningitis (arrow), and perivascular inflammation (paired arrows) and fewer T cell infiltration, compared with the Control group. The stained sections are representative of seven to nine mice per group.

Article Snippet: The CNS sections were incubated with antibodies against CD3 (T cell marker, 100-fold dilution, Biocare Medical, Pacheco, CA; antigen retrieval: 10 mM citrate buffer pH 6.0 at 120°C for 15 min), B220 (B cell marker, 300-fold dilution, eBioscience, San Diego, CA; no antigen retrieval), F4/80 (macrophage marker, 200-fold dilution, Bio-Rad, Hercules, CA; antigen retrieval: 100 μg/ml proteinase-K for 10 min), Ly-6G (neutrophil marker, 500-fold dilution, BD Bioscience, Franklin Lakes, NJ; no antigen retrieval), Foxp3 (Treg marker, 100-fold dilution, eBioscience; antigen retrieval: 10 mM citrate buffer pH 6.0 at 120°C for 15 min), and goat anti-mouse IgA-UNLB antibody (IgA-producing cell marker, 2000-fold dilution, SouthernBiotech, Birmingham, AL; no antigen retrieval) ( ).

Techniques: Control, Saline, MANN-WHITNEY, Staining