眼科新进展2017，Vol.37Issue(7)：605-609,5.DOI:10.13389/j.cnki.rao.2017.0154
TGF-β2刺激人Tenon囊成纤维细胞生物学行为改变及表达整合素连接激酶(ILK)的研究
Biological behavior changes and intergrin-linked kinase (ILK) expression after exposure to TGF-β2 in human Tenon fibroblasts
摘要
Abstract
Objective To investigate the biological behavior and expression of intergrin-linked kinase (ILK) after exposure to transforming growth factor beta 2 (TGF-β32) in human Tenon fibroblasts (HTFs).Methods The primary ceils of HTFs were cultured by tissue attached culture method and identified by immunofluorescence analysis with Vimentin and keratin.The proliferation levels of HTFs induced by different concentrations of TGF-β2 were analyzed by MTT.The α-smooth muscle actin (α-SMA) and ILK mRNA expression were analyzed by quantitative Real-time PCR.The protein expression of α-SMA,ILK and E-cadherin were analyzed by Western Blot,and the protein expression of α-SMA and E-cadherin were also analyzed by immunofluorescence staining.Results MTT analysis showed that the optical density levels of 5.0 μg · L-1 and 10.0 μg · L-1 TGF-β2 were significantly higher than those of 0.1 μg · L 1,1.0 μg · L-1 and the control after exposure for 48 hours and 72 hours,and all these optical density levels were significantly higher than that for 24 hours (all P ＜0.05).The expression of α-SMA and ILK mRNA increased significantly when cells were treated with 5.0 μg · L-1 TGF-β2 for 48 hours in comparison with the control group (all P ＜ 0.05).The protein of α-SMA,ILK and E-cadherin were expressed both in TGF-β2 treated groups and control group,and TGF-β2 up-regulated the expression of them.There were significant differences when compared with the control group (all P ＜ 0.05).Immunofluorescent staining showed that α-SMA and E-cadherin were detected in TGF-β2 treated groups.α-SMA expressed in cytoplasm,while E-cadherin both in cytoplasm and nucleus.Conclusion TGF-β2 can induce the proliferation,transdifferentiation and adhesion of HTFs,and up-regulate the expression of ILK in vitro,suggests that ILK may play a role in the process of scar formation after glaucoma filtration surgery.关键词
成纤维细胞/转化生长因子/整合素连接激酶/增殖/转分化Key words
fibroblasts/transforming growth factor/integrin-linked kinase/proliferation/transdifferentiation分类
医药卫生引用本文复制引用
崔丽珺,孙娜,田蕴霖,王群,裴澄,谢安明..TGF-β2刺激人Tenon囊成纤维细胞生物学行为改变及表达整合素连接激酶(ILK)的研究[J].眼科新进展,2017,37(7):605-609,5.基金项目
国家自然科学基金项目(编号:81470614、81300765、81070721) （编号:81470614、81300765、81070721）
西安交通大学基本科研业务费自由探索项目(编号:xjj2012062) （编号:xjj2012062）
西安交通大学医学院第一附属医院青年创新基金(编号:2010-858).National Natural Science Foundation of China (No:81470614,81300765,81070721) （编号:2010-858）
Fundamental Research Funds for the Central Universities sponsored by Xi'an Jiaotong University (No:xjj2012062) （No:xjj2012062）
Youth Foundation of the First Affiliated Hospital,Xi'an Jiaotong University (No:2010-858) （No:2010-858）
