摘要
目的探讨转化生长因子-β1(transforming growth factor-β1,TGF-β1)对人结膜下 Tenon 囊成纤维细胞(human subconjunctival Tenon's capsule fibroblast,HTCF)表型转化的作用及其信号转导通路。方法白内障手术中取人结膜下 Tenon 囊组织块培养成纤维细胞。用第5～9代细胞,以10μg/L TGF-β1诱导 HTCF48h,使其表型转化为肌成纤维细胞(myofibroblast,MF)。用蛋白质印迹免疫检测技术和免疫细胞化学技术鉴定细胞表型。在诱导后不同时间点(0、5、30、60、120、180min,12、24、36h),用蛋白质印迹免疫检测技术和免疫细胞化学技术测定smad2、AKT、ERK、P38蛋白表达。结果 10μg/L TGF-β1诱导 HTCF 后α-SMA 在24h 达峰值,蛋白表达在48h 最高;10μg/L TGF-β1 诱导 HTCF 后 smad2、AKT、P38表达具有时间双向性,smad2具有两个峰值。结论 TGF-β1能显著诱导 HTCF,使其表型转化为 MF,smad 信号通路参与了这一过程。(中国眼耳鼻喉科杂志,2007,7:14～16)
Purpose To explore the transforming effect on phenotype from the human subconjunctival Tenon＇ s capsule fibwblast （HTCF） to Myofibroblast （MF） induced by transforming growth factor β1（TGF-β1） and its signal pathway. Methods HTCF was derived and cultured from the Tenon＇ s capsule explant during cataract surgery. These HTCF were induced with TGF-β1 for 48 h. The phenotype of the HTCF was tmnsformed to MF that was verified by immunocytechemistry RT-PCR and western blotting technology. The level of α-smooth muscle-actin （α-SMA） and the expressions of smad2, AKT, ERK and P38 were measured at different time （0,5,30,60,120,180 min, 12,24,36 h） after induction with TGF-β1 at a level of 10 μg/L.Results The level of α-SMA reached to the peak 24 h and the expressino of the protein, 48 h after induction. The expression of smad2, AKT and P38 increased obviously after induction and there were two peaks of the expression of smad2. Conclusion TGF-β1 can induce phenotype transformation of HTCF to MF. Smad Signal pathway may be involved in this transforming course. （Chin J Ophthalmol and Otorhinolaryngol,2007,7：14-16）
出处
《中国眼耳鼻喉科杂志》
2007年第1期14-16,I0001,共4页
Chinese Journal of Ophthalmology and Otorhinolaryngology
基金
上海市科委重大课题(04JC14023)
