Pirfenidone attenuates IL-1¦Â-induced COX-2 and PGE₂ production in orbital fibroblasts through suppression of NF-¦ÊB activity

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• 作者：Youn-Hee Choi ; Keum Ok Back ; Hee Ja Kim ; Sang Yeul Lee ; Koung Hoon Kook
• 关键词：thyroid-associated ophthalmopathy ; orbital fibroblast ; pirfenidone ; COX-2 ; PGE2 ; IL-1¦Â ; NF-¦ÊB
• 刊名：Experimental Eye Research
• 出版年：2013
• 出版时间：August, 2013
• 年：2013
• 卷：113
• 期：Complete
• 页码：1-8
• 全文大小：1033 K

文摘

The aim of this study was to determine the effect of pirfenidone on interleukin (IL)-1¦Â-induced cyclooxygenase (COX)-2 and prostaglandin (PG)E₂ expression in orbital fibroblasts from patients with thyroid-associated ophthalmopathy (TAO). Primary cultures of orbital fibroblasts from patients with TAO (n?=?4) and non-TAO subjects (n?=?4) were prepared. The level of PGE₂ in orbital fibroblasts treated with IL-1¦Â in the presence or absence of pirfenidone was measured using an enzyme-linked immunosorbent assay. The effect of pirfenidone on IL-1¦Â-induced COX-2 expression in orbital fibroblasts from patients with TAO was evaluated by reverse transcription-polymerase chain reaction (PCR) and quantitative real-time PCR analyses, and verified by Western blot. Activation of nuclear factor-¦ÊB (NF-¦ÊB) was evaluated by immunoblotting for inhibitor of ¦ÊB (I¦ÊB)¦Á and phosphorylated I¦ÊB¦Á, and DNA-binding activity of p50/p65 NF-¦ÊB was analyzed by electrophoretic mobility shift assay. In addition, IL-1 receptor type 1 (IL-1R1) expression was assessed by RT-PCR in IL-1¦Â-treated cells with or without pirfenidone. Pirfenidone significantly attenuated IL-1¦Â-induced PGE₂ release in both TAO and non-TAO cells. IL-1¦Â-induced COX-2 mRNA and protein expression decreased significantly following co-treatment with pirfenidone. IL-1¦Â-induced I¦ÊB¦Á phosphorylation and degradation decreased in the presence of pirfenidone and led to decreased nuclear translocation and DNA binding of the active NF-¦ÊB complex. In our system, neither IL-1¦Â nor pirfenidone co-treatment influenced IL-1R1 expression. Our results suggest that pirfenidone attenuates the IL-1¦Â-induced PGE₂/COX-2 production in TAO orbital fibroblasts, which is related with suppression of the NF-¦ÊB activation.