Department of Orthopaedics, The Sixth Affiliated People's Hospital, Shanghai Jiaotong University School of Medicine, 600 Yishan Road, Shanghai 200233, China.
Eur J Pharmacol. 2012 Mar 5;678(1-3):1-5. doi: 10.1016/j.ejphar.2011.12.018. Epub 2011 Dec 23.
In the present study, the effects of celecoxib on proliferation, collagen expression, ERK1/2 and SMAD2/3 phosphorylation in NIH/3T3 fibroblasts were investigated. NIH/3T3 fibroblasts stimulated with fibroblast growth factor-2 (FGF-2) or transforming growth factor-β1 (TGF-β1) were examined in the presence of celecoxib. Proliferation was assessed by MTT assays; ERK1/2 expression and SMAD2/3 expression were assessed by quantitative RT-PCR and western blotting; ERK1/2 phosphorylation and SMAD2/3 phosphorylation were assessed by western blot analysis. The results indicated that celecoxib could suppress cell proliferation stimulated by FGF-2 (IC(50) FGF+group, 75±1.9μmol/l) and TGF-β1 (IC(50) TGF+group, 48±1.4μmol/l), by inhibiting ERK1/2 phosphorylation but not ERK1/2 expression. Celecoxib also suppressed collagen expression (0.35-fold COL3 and 0.43-fold COL1 at 320μmol/l celecoxib relative to the untreated control after stimulation with TGF-β1 for 3h, P<0.01), by inhibiting SMAD2/3 phosphorylation but not SMAD2/3 expression. The suppression of NIH/3T3 fibroblast proliferation and collagen expression upon stimulation by FGF-2 and TGF-β1 is likely a result of the inhibition of ERK1/2 and SMAD2/3 phosphorylation by celecoxib.
在本研究中，研究了塞来昔布对 NIH/3T3 成纤维细胞增殖、胶原表达、ERK1/2 和 SMAD2/3 磷酸化的影响。在存在塞来昔布的情况下，检查了由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。通过 MTT 测定法评估增殖；通过定量 RT-PCR 和 Western blot 评估 ERK1/2 表达和 SMAD2/3 表达；通过 Western blot 分析评估 ERK1/2 磷酸化和 SMAD2/3 磷酸化。结果表明，塞来昔布可抑制 FGF-2（IC（50）FGF+组，75±1.9μmol/l）和 TGF-β1（IC（50）TGF+组，48±1.4μmol/l）刺激的细胞增殖，通过抑制 ERK1/2 磷酸化而不是 ERK1/2 表达。塞来昔布还抑制胶原表达（在 TGF-β1 刺激 3 小时后，与未处理对照相比，在 320μmol/l 塞来昔布时 COL3 减少 0.35 倍，COL1 减少 0.43 倍，P<0.01），通过抑制 SMAD2/3 磷酸化而不是 SMAD2/3 表达。FGF-2 和 TGF-β1 刺激的 NIH/3T3 成纤维细胞增殖和胶原表达的抑制可能是塞来昔布抑制 ERK1/2 和 SMAD2/3 磷酸化的结果。
