TGF-β_1对甲状腺相关性眼病眼外肌成纤维细胞Smads基因表达的影响

Effects of transforming growth factor-β_1 on gene expression of Smads in orbital fibroblasts with thyroid-associated oph thalmolopathy

目的:测定转化生长因子-β1(transforming growth factor-β1,TGF-β1)对甲状腺相关性眼病(thyroid-associated ophthalmopathy,TAO)患者纤维化眼外肌来源的成纤维细胞(orbital fibroblasts,OF)细胞Smad3,Smad4,Smad7基因表达的影响。方法:采用实时荧光定量逆转录聚合酶连反应(real-time quantitative RT-PCR)观察5μg/L TGF-β1刺激OF在不同时间点(0h;15,30min;1,2,4h)表达Smad3 mRNA,Smad4 mRNA,Smad7 mRNA的变化。结果:5μg/L TGF-β1刺激OF 15min后,Smad3 mRNA的表达量已显著增加,为对照组的10.71倍,于1h达顶峰,为对照组的25.07倍(P<0.01),持续近2h,然后呈下降趋势,4h基本恢复正常;5μg/L TGF-β1刺激OF 15min后,Smad4 mRNA的表达量开始增加,为对照组的1.54倍,于1h达顶峰,为对照组的15.99倍(P<0.01),然后呈下降趋势,4h基本恢复正常;5μg/L TGF-β1刺激OF30min后,Smad7 mRNA的表达量明显开始增加,为对照组的3.21倍,持续至4h为对照组的14.66倍(P<0.01)。结论:TGF-β1可活化眼眶成纤维细胞Smads信号通路。在一定的时间范围内,诱导OF表达Smad3 mRNA和Smad4 mRNA呈先增加后下降的趋势,而Smad7 mRNA表达则呈现持续增加的趋势,说明TGF-β1/Smads这一纤维化相关的经典信号转导通路可能在TAO眼外肌纤维化机制中发挥了重要的作用。

AIM：To investigate the effects of transforming growth factor-β1 （TGF-β1 ） on gene expression of main components of Smads family including Smad3, Smad4 and Smad7 in orbital fibroblasts（OF） of fibrosis extraocular muscle with thyroid -associated ophthalmopathy（TAO）. METHODS. OF were treated with 5g/L TGF-β1 at different time points （0min, 15min, 30min,lh, 2h and 4h）, and real-time quantitative RT-PCR was performed to observe the effects of TGF- β1 on the expression of Smad3, Smad4 and Smad7 mRNA. RESULTS= The expression of Smad3 mRNA was 10.71 times to that of control group at 15min and 25.07 times at 1h（P〈0.01）; Smad4 mRNA was 1.54 times to that of control group at 15min and 15.99 times at lh（P〈0.01 ） ; Smad7 mRNA was 3.21 times to that of control group at 30rain and 14.66 times at 4h（ P〈0.01 ）. CONCLUSION - TGF- β up- regulate the expression of Smad3, Smad4, Smad7 mRNA in OF in a time dependent fashion, TGF-β1/Smad pathway may play an important role in the pathogenesis of extraocular muscle fibrosis with TAO.