Figure 8. RG1-MSC-CM promotes RIII recovery via increased release of VEGF and IL-6. (A) 8 selected cytokines in unconcentrated conditioned medium derived from BM-MSCs pre-activated by different concentration of RG1 (0.1 μM, 1 μM, 10 μM, 100 μM) were detected by ELISA. Data represent mean ± SD of at least three independent experiments. ##, P < 0.05 versus MSC-CM. (B) The effect of neutralization of 3 selected cytokines or addition of selected exogenous cytokines on apoptosis (left panel) and proliferation (right panel) of intestine in irradiated rats. The apoptosis and proliferation were evaluated by quantification of TUNEL-positive and PCNA-positive cell per 5 crypts 3 days after radiation, respectively. Data is reported as mean ± SD (n = 4~5). ##, P < 0.05 versus IR + DMEM-F12. **, P < 0.05 versus IR + RG1-MSC-CM. RMC: RG1-MSC-CM, 2NAs: anti-VEGF antibody + anti-IL-6 antibody. Scar bar 50 μm and 100 μm. (C) Pro/anti-inflammatory cytokines were extracted from jejunal protein of irradiated rats on day 3. The level of cytokines was determined by enzyme-linked immunosorbent assay (ELISA). (3~4 rats/group). **, P < 0.05 versus IR + DMEM-F12. ##, P < 0.05 versus IR + RG1-MSC-CM. (D) The effect of neutralization (left panel) of selected cytokines or addition of selected exogenous (right panel) cytokines on the survival of irradiated rats. Cumulative survival analyzed using the Kaplan-Meier method. P-values were determined by log-rank testing. The number of rats in each group is present in the parenthesis. (E) Mean survival time. Data represent the mean ± SD. **, P < 0.05 versus IR + DMEM-F12. ##, P < 0.05 versus IR + RG1-MSC-CM.