[1]崔丽珺,孙娜,田蕴霖,等.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.[doi:10.13389/j.cnki.rao.2017.0154]
　CUI Li-Jun,SUN Na,TIAN Yun-Lin,et al.Biological behavior changes and intergrin-linked kinase （ILK） expression after exposure to TGF-β2 in human Tenon fibroblasts[J].Recent Advances in Ophthalmology,2017,37(7):605-609.[doi:10.13389/j.cnki.rao.2017.0154]
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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/HTML
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《眼科新进展》[ISSN:1003-5141/CN:41-1105/R]

卷:
37卷
期数:
2017年7期
页码:
605-609
栏目:
实验研究
出版日期:
2017-07-05

文章信息/Info

Title:
Biological behavior changes and intergrin-linked kinase （ILK） expression after exposure to TGF-β2 in human Tenon fibroblasts
作者:
崔丽珺; 孙娜; 田蕴霖; 王群; 裴澄; 谢安明
710061　陕西省西安市，西安交通大学第一附属医院眼科（崔丽珺，王群，裴澄，谢安明）；710004　陕西省西安市，陕西省眼科诊疗中心，西安市第四医院眼科，西安交通大学医学院附属广仁医院眼科（孙娜，田蕴霖）
Author(s):
CUI Li-Jun; SUN Na; TIAN Yun-Lin; WANG Qun; PEI Cheng; XIE An-Ming
Department of Ophthalmology，the First Affiliated Hospital of Xi’an Jiaotong University（CUI Li-Jun，WANG Qun，PEI Cheng，XIE An-Ming），Xi’an 710061，Shaanxi Province，China；Department of Shaanxi Ophthalmic Medical Center，Xi’an No.4 Hospital，Affiliated Guangren Hospital，Xi’an Jiaotong University（SUN Na，TIAN Yun-Lin），Xi’an 710004，Shaanxi Province，China
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
Keywords:
fibroblasts; transforming growth factor; integrin-linked kinase; proliferation; transdifferentiation
分类号:
R775
DOI:
10.13389/j.cnki.rao.2017.0154
文献标志码:
A
摘要:
目的　探讨转化生长因子β2（transforming growth factor beta 2，TGF-β2）对体外培养的人Tenon囊成纤维细胞（human Tenon fibroblasts，HTFs）生物学行为及表达整合素连接激酶（integrin-linked kinase，ILK）的影响。方法　组织块贴壁法培养原代HTFs，免疫荧光染色鉴定波形蛋白和角蛋白。MTT法检测不同浓度TGF-β2对HTFs细胞的促增殖作用，RT-PCR检测TGF-β2作用前后α-平滑肌肌动蛋白（smooth muscle actin，SMA）、ILK mRNA的表达，Western Blot检测α-SMA、ILK、E-cadherin蛋白的表达，细胞免疫荧光染色检测α-SMA、E-cadherin蛋白的表达。结果　MTT检测结果显示，TGF-β2作用48 h、72 h，5.0 μg·L-1及10.0 μg·L-1诱导下的OD值显著高于0.1 μg·L-1、1.0 μg·L-1组及空白对照组（均为P＜0.05），5.0 μg·L-1及10.0 μg·L-1作用48 h及72 h的OD值显著高于24 h（均为P＜0.05）。RT-PCR检测结果显示，5.0 μg·L-1TGF-β2诱导48 h后，α-SMA mRNA及ILK mRNA水平和空白对照组相比明显上调，差异均有统计学意义（均为P＜0.05）。Western Blot检测结果显示，ILK、α-SMA及E-cadherin在空白对照组和5.0 μg·L-1TGF-β2处理组均有表达，但TGF-β2能够明显上调三者的表达，5.0 μg·L-1TGF-β2处理48 h组与空白对照组差异均有统计学意义（均为P<0.05）。细胞免疫荧光染色结果显示，TGF-β2处理组α-SMA、E-cadherin均有荧光表达，其中α-SMA表达在细胞质中，而E-cadherin在细胞质和细胞核中均有表达。结论　TGF-β2可以诱导体外培养HTFs增殖，转分化及黏附性增强，同时促进ILK的高表达，提示ILK在青光眼术后瘢痕化过程中可能也起到一定作用。
Abstract:
Objective　To investigate the biological behavior and expression of intergrin-linked kinase （ILK） after exposure to transforming growth factor beta 2 （TGF-β2） in human Tenon fibroblasts （HTFs）.Methods　The primary cells 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.

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备注/Memo

备注/Memo:
国家自然科学基金项目（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）；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）
更新日期/Last Update: 2017-08-02