Figure 5.

miR-200a overexpression attenuated DSS-induced colonic injury in vitro. (A) The expression of miR-200a in cells transfected with the miR-200a mimic or the miR-scramble control for 48 h. (B–E) The levels of inflammatory cytokines (IL-1β, TNF-α, IL-6, IL-10) were analyzed by ELISA kits in the NCM460 cells. (F) The production of MDA in Enterocytes was detected by commercial kits. (G) The activity of ROS in Enterocytes was detected by commercial kits. (H, I) The expression of 4-HNE and GSH/GSSG in Enterocytes was detected by commercial kits. (J–M) Representative immunoblot bands for the Nrf2, Keap1, and NQO1 proteins in Enterocytes. GAPDH was used as a loading control. (N) The mRNA levels of the Nrf2-regulated gene were detected by qRT-PCR in the NCM460 cells. (O) Cell viability detected by MTT in Enterocytes from different experimental groups. (P, R) Representative images of TUNEL staining in the NCM460 cells from different experimental groups. (Original magnifications, ×200, scale bars =50 μm);. The images of TUNEL positive for quantification were analyzed using Image J software. (Q, S) Representative images of the percentage of apoptosis were examined by flow cytometry. Data are provided as mean ± SEM and reflect three experiments, analyzed using one-way ANOVA with Tukey post-hoc analysis. ^∗P < 0.05; ns, not significant.