Figure 4
CKD-506 decreased the levels of Th1/17-secreted pro-inflammatory cytokines and decreased the proliferation of myelin oligodendrocyte glycoprotein35–55 (MOG35–55)-stimulated splenocytes. Mice induced using MOG35–55 and pertussis toxin were sacrificed by cervical dislocation on day 12 post-induction and the splenocytes were isolated. Mice were not treated with any drugs, including vehicle. CCK8 solution was used to examine the proliferation of the splenocytes incubated with the MOG35–55 peptide (10, 20, and 40 μg/mL), vehicle, and CKD-506 (1 and 3 μM) for 72 h at 37 °C. The absorbance values of the mixtures were analyzed 3 h later. The data were obtained from three independent experiments with three replicates (a). Televels of IFN-γ, IL-12, IL-17A, IL-1β, IL-4, and TNF-α in the culture supernatant of the splenocytes stimulated with MOG35–55 (20 μg/mL) for 72 h at 37 °C and those in the culture supernatant of stimulated splenocytes co-incubated with CKD-506 (3 μM) were examined (n = 9 per group) (b). Data are presented as mean ± SD. Two-way ANOVA, followed by Bonferroni’s post-hoc test for (a); one-way ANOVA, followed by Dunnett’s post-hoc (b). #p < 0.05, and ###p < 0.001, non-stimulated groups versus MOG-stimulated group (vehicle group); *p < 0.05, **p < 0.01, and ***p < 0.001, drug-treated group versus vehicle group. No (gray color), vehicle treatment; low (light red), 1 μM CKD-506 treatment; high (dark red), 3 μM CKD-506 3 μM treatment.
