Department of Orthopaedics, The Sixth Affiliated People's Hospital, Shanghai Jiaotong University, Shanghai 200233, PR China.
Mol Med Rep. 2012 Mar;5(3):827-31. doi: 10.3892/mmr.2011.722. Epub 2011 Dec 19.
This study aimed to investigate whether celecoxib suppresses fibroblast proliferation and collagen expression by inhibiting extracellular signal-regulated kinase 1/2 (ERK1/2) and SMAD2/3 phosphorylation. Celecoxib was added to NIH/3T3 fibroblasts stimulated by fibroblast growth factor-2 (FGF-2) or transforming growth factor-β1 (TGF‑β1). NIH/3T3 fibroblast proliferation and viability were assessed by MTT assays; ERK1/2 expression and SMAD2/3 expression were assessed by quantitative RT-PCR and Western blot analysis. The results indicated that celecoxib suppressed cell proliferation (IC50 FGF+ group, 75 ± 1.9 µmol/l) stimulated by FGF-2, and also inhibited cell viability (IC50 FGF- group, 252 ± 2.3 µmol/l) by inhibiting ERK1/2 phosphorylation but not ERK1/2 expression. In addition, celecoxib treatment led to the apoptosis of NIH/3T3 fibroblasts (IC50 FGF- group, 35 ± 1.4 µmol/l). Celecoxib also suppressed collagen expression (0.35-fold COL3 and 0.43-fold COL1 with 320 µmol/l celecoxib relative to the untreated group following stimulation for 3 h, p<0.01) when stimulated by TGF‑β1, by inhibiting SMAD2/3 phosphorylation but not SMAD2/3 expression. Celecoxib is capable of inhibiting ERK1/2 and SMAD2/3 phosphorylation, which is responsible for NIH/3T3 fibroblast proliferation and collagen expression.
本研究旨在探讨塞来昔布是否通过抑制细胞外信号调节激酶 1/2（ERK1/2）和 SMAD2/3 磷酸化来抑制成纤维细胞增殖和胶原表达。向成纤维细胞生长因子-2（FGF-2）或转化生长因子-β1（TGF-β1）刺激的 NIH/3T3 成纤维细胞中加入塞来昔布。通过 MTT 测定法评估 NIH/3T3 成纤维细胞增殖和活力；通过定量 RT-PCR 和 Western blot 分析评估 ERK1/2 表达和 SMAD2/3 表达。结果表明，塞来昔布抑制 FGF-2 刺激的细胞增殖（FGF+组 IC50 为 75±1.9µmol/l），并通过抑制 ERK1/2 磷酸化而不是 ERK1/2 表达来抑制细胞活力（FGF-组 IC50 为 252±2.3µmol/l）。此外，塞来昔布处理导致 NIH/3T3 成纤维细胞凋亡（FGF-组 IC50 为 35±1.4µmol/l）。塞来昔布还通过抑制 SMAD2/3 磷酸化而不是 SMAD2/3 表达，抑制 TGF-β1 刺激的胶原表达（320µmol/l 塞来昔布处理 3 小时后 COL3 降低 0.35 倍，COL1 降低 0.43 倍，与未处理组相比，p<0.01）。塞来昔布能够抑制 ERK1/2 和 SMAD2/3 磷酸化，这是 NIH/3T3 成纤维细胞增殖和胶原表达的原因。
