Effects of R723 on inflammation-related gene expression and microgliosis in spinal cord of mSOD1
transgenic mice. (A) R723-treated mSOD1G93A mice had reduced number of lectin-positive microglia in the spinal cord compared with vehicle-treated controls. Lumbar sections of the spinal cord were stained with fluorescein isothiocyanate (FITC)-conjugated tomato lectin. Scale bar = 100 μm. Data are representative of three animals. (B, C) Quantitative RT-PCR analyses in spinal cords of R723-treated mSOD1G93A mice and vehicle-treated controls (120 days old) were performed (n = 4 in each group). The expression levels of INF-γ and iNOS were significantly reduced in the R723-treated group (P = 0.0495 for each experiment). (D) Immunohistochemical analysis showed R723 treatment for 30 days had suppressed the expression level of iNOS in the spinal cords of mSOD1G93A mice. Scale bar = 100 μm. Data are representative of three animals. (E) Quantitative RT-PCR analyses in spinal cords of R723 treated mSOD1G93A mice and vehicle-treated controls (120 days old) were performed (n = 4 in each group). Relative mRNA expression is shown for TNF, MCP1, Il-12b, Il-6, Il-1b, NOX2, and Ly6c, which are related to M1 macrophages/microglia, and for Il-4, Arg1, Ym1, Il-4, EPO and CSF3, which are related to M2 macrophages/microglia. There were no significant differences in the expression levels of these molecules between two groups after the correction of multiple comparisons. (F) Quantitative RT-PCR analysis revealed that R723 had suppressed the expression of Retnla after 30 days of treatment in the spinal cords of mSOD1G93A mice (P = 0.0495, n = 4 in each group). Data are expressed as means ± SEM. *P < 0.05, Mann-Whitney U-test.