[1]李静,谢安明,张坚,等. Bevacizumab对人Tenon囊成纤维细胞转分化的抑制作用[J].眼科新进展,2015,35(7):615-618.[doi:10.13389/j.cnki.rao.2015.0167]
　LI Jing,XIE An-Ming,ZHANG Jian,et al. Inhibitive effects of Bevacizumab on transdifferentiation of human Tenon capsule fibroblasts[J].Recent Advances in Ophthalmology,2015,35(7):615-618.[doi:10.13389/j.cnki.rao.2015.0167]
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 Bevacizumab对人Tenon囊成纤维细胞转分化的抑制作用
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《眼科新进展》[ISSN:1003-5141/CN:41-1105/R]

卷:
35卷
期数:
2015年7期
页码:
615-618
栏目:
实验研究
出版日期:
2015-07-05

文章信息/Info

Title:
 Inhibitive effects of Bevacizumab on transdifferentiation of human Tenon capsule fibroblasts
作者:
 李静; 谢安明; 张坚; 王建萍; 薛雨顺
 710068　陕西省西安市，陕西省人民医院眼科
Author(s):
 LI Jing ; XIE An-Ming ; ZHANG Jian ; WANG Jian-Ping ; XUE Yu-Shun
 Department of Ophthalmology , Shaanxi Provincial People ’ s Hospital , Xi ’ an 710068 . Shaanxi Province , China
关键词:
 ; 人Tenon囊成纤维细胞; Bevacizumab; 转分化; 转化生长因子; 青光眼
Keywords:
 ; human Tenon ’ s capsule fibroblasts ;  Bevacizumab ;  transdifferentiation ;  transforming growth factor ;  glaucoma
DOI:
10.13389/j.cnki.rao.2015.0167
文献标志码:
A
摘要:
 目的　观察Ｂｅｖａｃｉｚｕｍａｂ（贝伐单抗）对转化生长因子（ｔｒａｎｓｆｏｒｍｉｎｇｇｒｏｗｔｈｆａｃｔｏｒ，ＴＧＦ）-β２诱导下的人Ｔｅｎｏｎ囊成纤维细胞转分化的抑制作用，探讨抗新生血管药物抑制青光眼术后滤过泡瘢痕化的机制。方法　用含体积分数１０％胎牛血清的ＤＭＥＭ高糖型培养基对人Ｔｅｎｏｎ囊成纤维细胞株进行体外常规培养和传代。实验分为空白对照组、ＴＧＦ-β２处理组（加入终浓度为１０μｇ?Ｌ－１的ＴＧＦ-β２ＤＭＥＭ完全培养基）及Ｂｅｖａｃｉｚｕｍａｂ干预组（加入１０μｇ?Ｌ－１的ＴＧＦ-β２和１．０ｇ?Ｌ－１的ＢｅｖａｃｉｚｕｍａｂＤＭＥＭ完全培养基），置于３７℃、体积分数５％二氧化碳培养箱中培养４８ｈ，并分别应用细胞免疫荧光染色技术和ＷｅｓｔｅｒｎＢｌｏｔ实验检测Ｂｅｖ-ａｃｉｚｕｍａｂ对ＴＧＦ-β２刺激下人Ｔｅｎｏｎ囊成纤维细胞α-平滑肌肌动蛋白（α-ｓｍｏｏｔｈｍｕｓｃｌｅａｃｔｉｎ，α-ＳＭＡ）表达的影响。结果　α-ＳＭＡ蛋白主要表达在细胞质中，免疫荧光染色结果显示ＴＧＦ-β２处理组可以见到α-ＳＭＡ的荧光表达，而Ｂｅｖａｃｉｚｕｍａｂ干预组α-ＳＭＡ蛋白表达受到抑制。ＷｅｓｔｅｒｎＢｌｏｔ结果显示空白对照组、ＴＧＦ-β２ 处理组及Ｂｅｖａｃｉｚｕｍａｂ干预组α-ＳＭＡ蛋白相对表达量分别为０．６３０±０．０３８、１．１３０±０．０７１和０．３４０±０．０３３，Ｂｅｖａｃｉｚｕｍａｂ干预组α-ＳＭＡ蛋白相对表达量低于空白对照组和ＴＧＦ-β２处理组（均为Ｐ＜０．０５）。结论　Ｂｅｖａｃｉｚｕｍａｂ可明显抑制ＴＧＦ-β２诱导下人Ｔｅｎｏｎ囊成纤维细胞α-ＳＭＡ蛋白的表达，抑制成纤维细胞表型转化。
Abstract:
 Objective To investigate the inhibitive effects of Bevacizumab on transdifferentiation of human Tenon ’ s capsule fibroblasts induced by transforming growth factor-β2 ( TGF-β2 ) in vitro , and explore the mecharusm of anti-angiogenesis drugs inhibiting bleb scar after glaucoma filtering surgery. Methods Human Tenon ’ s capsule fibroblasts cell line were cultured and passaged in DMEM/High glucose medium with 10% fetal bovine serum. The cells were divided into blank control group, TGF-β2 group and TGF-β2 + Bevacizumab group. Medium containing a final concentration of 10 vg . L -l TGF-β2 was added into TGF-β2 group and medium containing a final concentration of 10 pdg . L-1 TGF-β2 plus l. 0 g . L-l Bevacizumab was added into TGF-β2 + Bevacizumab group which were used into the 37 0C ,5% carbon dioxide cultured 48 hours. Immunofluorescence staining and Western Blot were used to measure the expression of d一smooth muscle actin (d—SMA) protein after culture for 48 hours with Bevacizumab. Results The expression of d—SMA protein mainly located in human Tenon fibroblast cytoplasm. Immunofluorescence staining showed that TGF-β2 up-regulated the expression of d—SMA protein. while Bevacizumab significantly inhibited the expression of dSMA protein. Western Blot results showed that the relative d—SMA protein expression of blank control group, TGF-β2 treatment group and TGF-βz + Bevacizumab treatment group were 0. 630 + 0. 038 . 1. 130 + 0. 071 and 0. 340 + 0. 033 . TGF-βz + Bevacizumab treatment group was lower than blank control group and TGF-β2 treatment group ( all P < 0. 05 ) . Conclusion Bevacizumab can obviously inhibit the expression ofd一SMA protein of human Tenon capsule fibroblasts, and inhibit fibroblast phenotype transformation.

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备注/Memo

备注/Memo:
 陕西省自然科学基础研究计划项目资助（编号：２０１４ＪＭ２-８１９０）
更新日期/Last Update: 2015-07-03