Fig 5. Effect of IL-1β on the NF-κB, p38, and ERK pathways in orbital fibroblasts, and the role of the NF-κB, p38, and ERK pathways in IL-1β-induced IL-8 expression in orbital fibroblasts.

(A) Orbital fibroblasts were treated with IL-1β (10 ng/mL) for 10–120 minutes. At the designated times, the degradation of IκBα and the phosphorylation of p38 and ERK were confirmed by Western blot analysis. Similar results were obtained in three independent experiments with orbital fibroblasts of passages (P)7 and P10 from TAO patient #53. (B) Orbital fibroblasts were pre-treated with inhibitors, BAY 11–7085 for NF-κB, SB 203580 for p38, or PD 98059 for ERK for 1 hour, followed by treatment with IL-1β (10 ng/mL) for 24 hours. IL-8 mRNA level was determined by quantitative real time RT-PCR. Similar results were obtained in three independent experiments with orbital fibroblasts of P9 and P10 from TAO patient #53. (C) Orbital fibroblasts were treated with BAY 11–7085, SB 203580, or PD 98059 for 24 hours. Cell viability was measured using the MTT reduction assay. Similar results were obtained in three independent experiments with orbital fibroblasts of P8 and P10 from TAO patient #53. *p < 0.05 compared with cells treated with the same concentration of IL-1β as calculated by one-way ANOVA. P, number of passages.