Figure 5From: Anti-invasive effects of Celastrus Orbiculatus extract on interleukin-1 beta and tumour necrosis factor-alpha combination-stimulated fibroblast-like synoviocytes NF-κB mediates IL-1β and TNF-α-induced MMP-9 expression. (A) FLSs were pretreated with BAY (5, 10 μM) for 1 h and then stimulated with IL-1β (10 ng/ml) and TNF-α (10 ng/ml) for 48 h. MMP-9 mRNA expression in cells was analysed by qRT-PCR. GAPDH was used as an internal control. The histogram shows the mRNA levels from three independent experiments. (B) and (C) FLSs were pretreated with BAY (5, 10 μM) for 1 h and then stimulated with IL-1β (10 ng/ml) and TNF-α (10 ng/ml). After 48 h, conditioned media was collected and gelatin zymography, western blotting or ELISA was performed. (D) FLSs were transfected with pGL2-MMP-9WT reporter plasmids and then cultured with BAY (5, 10 μM) with or without IL-1β (10 ng/ml) and TNF-α (10 ng/ml) for 48 h. Luciferase activity in the cell extracts was determined. The data are presented as mean ± SD of triplicate experiments. ** P < 0.01 versus normal control group, ◆ P < 0.05, ◆◆ P < 0.01 versus IL-1β and TNF-α combination-treated group.Back to article page