[1]任师杰,张凤妍,祁颖,等.法舒地尔对转化生长因子-β2诱导的人晶状体上皮细胞迁移及细胞外基质合成的影[J].眼科新进展,2014,34(5):418-421.[doi:10.13389/j.cnki.rao.2014.0115]
　REN Shi-Jie,ZHANG Feng-Yan,Ql Ying,et al.Effects of fasudil on human lens epithelial cell migration and extracellular matrix synthesis induced by transforming growth factor- β2[J].Recent Advances in Ophthalmology,2014,34(5):418-421.[doi:10.13389/j.cnki.rao.2014.0115]
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法舒地尔对转化生长因子-β2诱导的人晶状体上皮细胞迁移及细胞外基质合成的影
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《眼科新进展》[ISSN:1003-5141/CN:41-1105/R]

卷:
34卷
期数:
2014年5期
页码:
418-421
栏目:
实验研究
出版日期:
2014-05-05

文章信息/Info

Title:
Effects of fasudil on human lens epithelial cell migration and extracellular matrix synthesis induced by transforming growth factor- β2
作者:
任师杰; 张凤妍; 祁颖; 崔琨明
４５００５２河南省郑州市， 郑州大学第一附属医院眼科
Author(s):
REN Shi-Jie ;  ZHANG Feng-Yan ;  Ql Ying ;  CUI Kun-Ming
关键词:
法舒地尔; 后发性白内障; 晶状体上皮细胞; 细胞外基质
Keywords:
fasudil;  posterior capsule opacification;  lens epithelial cells ;  extracellular matrix
DOI:
10.13389/j.cnki.rao.2014.0115
文献标志码:
A
摘要:
目的　探讨法舒地尔对转化生长因子-β２（ｔｒａｎｓｆｏｒｍｉｎｇｇｒｏｗｔｈｆａｃｔｏｒ-β２，ＴＧＦ-β２）诱导的人晶状体上皮细胞（ｌｅｎｓｅｐｉｔｈｅ- ｌｉａｌｃｅｌｌｓ，ＬＥＣｓ）ＨＬＥ-Ｂ３迁移及细胞外基质合成的影响。方法　将培养的ＨＬＥ-Ｂ３分为以下４组：Ａ组：含体积分数１０％胎牛血清的完全培养基；Ｂ组：含１０μｇ?Ｌ－１ＴＧＦ-β２的完全培养基；Ｃ组：含１５μｍｏｌ?Ｌ－１法舒地尔的完全培养基；Ｄ组：含１０μｇ? Ｌ－１ＴＧＦ-β２＋１５μｍｏｌ?Ｌ－１法舒地尔的完全培养基。ＣＣＫ-８法检测不同浓度的法舒地尔对细胞增殖的影响，Ｔｒａｎｓｗｅｌｌ小室迁移实验检测各组细胞迁移能力，ＥＬＩＳＡ法检测各组细胞上清Ｉ型胶原（ｃｏｌｌａｎｇｅ-Ｉ，ＣＯＬ-Ｉ）、纤维连接蛋白（ｆｉｂｒｏｎｅｃｔｉｎ，ＦＮ）的表达。结果　随着法舒地尔干预浓度的增加，对ＨＬＥ-Ｂ３细胞的增殖抑制作用逐渐增强，各浓度组间差异有统计学意义（Ｐ＝０．０００）。随着法舒地尔作用时间延长，对ＨＬＥ-Ｂ３细胞增殖的抑制作用也逐渐增强，各组间差异有统计学意义（Ｐ＝０．０００）。Ｔｒａｎｓｗｅｌｌ小室迁移实验显示，４８ｈ时Ａ组、Ｂ组、Ｃ组、Ｄ组细胞穿过聚碳酸酯嵌套膜的个数分别为（２９．１７±１．１７）个、（８７．６０±５．３１）个、（１５．６０±１．０７）个、（４４．６１±２．０６）个，各组间差异有统计学意义（Ｆ＝１１１．６１４，Ｐ＝０．０００）；Ｂ组较Ａ组、Ｄ组较Ｃ组迁移细胞明显增多（均为Ｐ＝０．０００）；Ｄ组较Ｂ组、Ｃ组较Ａ组迁移细胞明显减少（Ｐ＝０．０００、Ｐ＝０．００３），但Ｄ组较Ａ组、Ｂ组较Ｃ组细胞迁移数量明显增多（Ｐ＝０．００３、Ｐ＝０．０００）。各组细胞上清液中ＦＮ、ＣＯＬ-Ｉ的含量于１２ｈ开始升高，２４ｈ、３６ｈ时持续升高，４８ｈ 时增高明显，各时间点各组间以及组内各时间点间差异均有统计学意义（均为Ｐ＜０．０５）。与Ａ组相比，各时间点Ｂ组ＦＮ和ＣＯＬ-Ｉ蛋白的表达明显增加（均为Ｐ＜０．０５）；各时间点Ｄ组较Ｂ组显著下降（均为Ｐ＜０．０５）；但Ｃ组与Ａ组比较差异均无统计学意义（均为Ｐ＞０．０５）；Ｄ组与Ａ组、Ｃ组与Ｂ组、Ｃ组与Ｄ组比较，ＣＯＬ-Ｉ、ＦＮ差异均有统计学意义（均为Ｐ＜０．０５）。结论　法舒地尔可抑制ＴＧＦ-β２诱导的细胞迁移和细胞外基质的合成，可能在后发性白内障的防治过程中发挥作用。
Abstract:
Objective To investigate the effects of fasudil on human lens epithelial cell ( HLEC)-B3 nugration and extracellular matrix synthesis induced by transforming growth factor-β2 ( TGF-β2 ) . Methods The cultured HLEC-B3 were divided into 4 groups : group A was the control group,the cells in group B were exposed in 10 μg . L-l TGF-β2 , group C in 15 μmol . L-I fasudil and group D in 15 μmol . L-l fasudil + 10 μg . L-l TGF-β2. Cell proliferation was assessed by CCK-8 assay, and the mobility of HLE-B3 was evaluated by Transwell assay. Then ELISA method was used to check the protein content of the fibronectin ( FN) and the collagen-l ( COL-I) in the supernatant. Results After treatment with different concentrations of fasudil, the proliferation of HLE-B3 was significantly inhibited, there was statistical difference among each group (P = 0. 000) . Fasudil inhibited HLE-B3 proliferation in a time dependent manner , there was statistical difference among different time points (P = 0. 000 ) . Transwell migration assay showed that the number of migration cells across the polycarbonate membrane at 48 hours in group A,B,C.D were 29. 17 + 1. 17 ,87. 60 +5. 31 . 15. 60 + 1. 07 ,44. 61 +2. 06 . respectively,there was statistical difference among each group ( F = 111. 614, P = 0. 000) , group B was obvious more than group A, and group D was obvious more than group C ( all P = 0. 000) , group D was obvious less than group B, group C was obvious less than group A (P = 0. 000.0. 003 ) , but group D was obvious more than group A, group B was more than group C (P = 0. 003 ,0. 000 ) . The contents of FN and COL-I in supernatant of each group increased from 12 hours .sustained at 24 hours and 36 hours. obvious increased at 48 hours .there were statistical differences among different groups at different time points and among different time points of different groups ( all P < 0. 05 ) . Compared with group A,the expression of FN and COL-I in group B at different time points obvious increased ( all P < 0. 05) , group D were obvious less than group B at different time points ( all P < 0. 05 ) . but there was no statistical difference between group C and group A ( all P > 0. 05) . and there were statistical differences between group A and D, group C and B, group C and D ( all P < 0. 05 ) . Conclusion Fasudil can inhibit the HLEC migration and extracellular matrix synthesis induced by transforming growth factor-β2 , which may provide new options for prevention and treatment of posterior capsule opacification.

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备注/Memo

备注/Memo:
郑州大学研究生科学研究基金项目（编号：１２Ｙ０３４０４）
更新日期/Last Update: 2014-05-04