Iguratimod Inhibits the Aggressiveness of Rheumatoid Fibroblast-Like Synoviocytes

Figure 4

Iguratimod suppressed the migration and invasion of RA-FLSs by blocking MAPK signaling. (a, c) RA-FLS monolayers were scratched with a sterile pipette tip. Each well was treated with 2% FBS and iguratimod at various concentrations. Microscopy images were acquired at 48 h after wounds were created. Representative microscopy images from the wound assay were shown. (b, d) RA-FLSs were starved in serum-free DMEM for 24 h, FLSs (, 200 μl) were seeded in the upper chambers, treated with different concentrations of iguratimod for 24 h, and then a transwell assay was performed. The data shown were the for three independent experiments (totally 6 RA-FLSs lines). (e) RA-FLSs were cultured in the presence of TNF-α (10 ng/ml) with or without iguratimod or DMSO at indicated times. Protein lysates obtained from equal numbers of RA-FLSs were analyzed by western blot. Total JNK, P38, ERK, phospho-JNK, phospho-P38, and phospho-ERK, as well as GAPDH, were detected. (f) RA-FLSs were cultured in the presence of TNF-α (10 ng/ml) with or without iguratimod or DMSO at indicated times. Protein lysates obtained from equal numbers of RA-FLSs were analyzed by western blot. Protein pATF-2, total ATF-2, ELK-1, and GAPDH were detected. The experiments were repeated four times (totally 4 RA-FLSs lines). The representative plots were shown. GAPDH was used as endogenous control, and relative expression of each protein is shown as a protein/GAPDH ratio. The summarized data were shown. The experiments were repeated three times. The data were analyzed using one-way ANOVA followed by Turkey’s test. , versus the control group.

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