[1]朱江,许治国,任梅,等.BAMBI过表达抑制缺氧诱导的恒河猴视网膜血管内皮细胞增殖和迁移[J].眼科新进展,2016,36(5):418-422.[doi:10.13389/j.cnki.rao.2016.0112]
　ZHU Jiang,XU Zhi-Guo,REN Mei,et al.Inhibitive effects of BAMBI overexpression on retinal vascular endothelial cell proliferation and migration induced by hypoxia in Rhesus monkey[J].Recent Advances in Ophthalmology,2016,36(5):418-422.[doi:10.13389/j.cnki.rao.2016.0112]
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BAMBI过表达抑制缺氧诱导的恒河猴视网膜血管内皮细胞增殖和迁移
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《眼科新进展》[ISSN:1003-5141/CN:41-1105/R]

卷:
36卷
期数:
2016年5期
页码:
418-422
栏目:
实验研究
出版日期:
2016-05-05

文章信息/Info

Title:
Inhibitive effects of BAMBI overexpression on retinal vascular endothelial cell proliferation and migration induced by hypoxia in Rhesus monkey
作者:
朱江; 许治国; 任梅; 党晓洁
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
Author(s):
ZHU Jiang; XU Zhi-Guo; REN Mei; DANG Xiao-Jie
Shaanxi Ophthalmic Medical Center , Xi ’an No. 4 Hospital , Affiliated Guangren Hospital , School of Medicine , Xi ’ an Jiaotong University , Xi ’ an 710004 ,Shaanxi Province , China
关键词:
视网膜血管内皮细胞; 缺氧; BAMBI; 增殖; 迁移
Keywords:
retinal vascular endothelial cell;  hypoxia ;  BAMBI ;  proliferation ;  migration
DOI:
10.13389/j.cnki.rao.2016.0112
文献标志码:
A
摘要:
目的　探讨过表达激活素膜结合抑制剂（ｂｍｐａｎｄａｃｔｉｖｉｎｍｅｍｂｒａｎｅ-ｂｏｕｎｄｉｎｈｉｂｉｔｏｒ，ＢＡＭＢＩ）对缺氧诱导的恒河猴视网膜血管内皮细胞（ＲＦ／６Ａ）增殖和迁移的影响。方法　将ＲＦ／６Ａ细胞分为四组：正常对照组、缺氧组、缺氧＋ｐＦＬＡＧ组和缺氧＋ＢＡＭＢＩ组。Ｗｅｓｔｅｒｎｂｌｏｔ检测各组细胞中ＢＡＭＢＩ的表达，倒置显微镜观察各组细胞形态和密度，ＣＣＫ-８法和流式细胞仪分别检测各组细胞的增殖和周期情况，Ｔｒａｎｓｗｅｌｌ检测细胞的迁移，Ｗｅｓｔｅｒｎｂｌｏｔ检测转化生长因子β１（ｔｒａｎｓｆｏｒｍｉｎｇｇｒｏｗｔｈｆａｃｔｏｒ-β１，ＴＧＦ-β１）和血管内皮生长因子（ｖａｓｃｕｌａｒｅｎｄｏｔｈｅｌｉａｌｇｒｏｗｔｈｆａｃｔｏｒ，ＶＥＧＦ）的表达。结果　与对照组相比，缺氧组细胞数目明显增多，增殖增加，培养１２０ｈ后，对照组细胞增殖倍数为５．００±０．２８，缺氧组为７．６４±０．３２（Ｐ＜０．００１）；缺氧组Ｓ期进程显著加快，占（１０．４４±２．６１）％，对照组Ｓ期占（２０．７９±２．２３）％（Ｐ＜０．０５）；细胞迁移增加，缺氧组细胞数目为３３．８０±４．２０，对照组为８．３５±２．５０（Ｐ＜０．０５），ＴＧＦ-β１和ＶＥＧＦ的表达明显增加（Ｐ＜０．０５）。缺氧＋ＢＡＭＢＩ组：缺氧处理的细胞转染ｐＦＬＡＧ-ＢＡＭ-ＢＩ后，与缺氧组相比，ＢＡＭＢＩ蛋白表达显著上调，细胞数目明显减少，增殖显著降低，培养１２０ｈ后，增殖倍数为５．５３±０．２７（Ｐ＜０．００１），细胞Ｓ期受到阻滞，占（１８．７５±２．９２）％，迁移显著下降，数目为１３．００±２．８０（Ｐ＜０．０５），ＴＧＦ-β１和ＶＥＧＦ的表达明显下调（Ｐ＜０．０５）。结论　过表达ＢＡＭＢＩ可抑制缺氧诱导的ＲＦ／６Ａ细胞的增殖和迁移，进而有望抑制视网膜血管新生。
Abstract:
Objective To explore the effects of bmp and activin membranebound inhibitor ( BAMBI) overexpression on retinal vascular endothelial cell ( RF/6A) proliferation and migration induced by hypoxia in thesus monkey. Methods Cell hypoxia model was established and cells were divided into four groups :Control group , hypoxia group, hypoxia + pFLAG group and hypoxia + BAMBI group. The expression of BAMBI was measured by Western blot. Cell morphology and density were observed under inverted microscope. Cell proliferation and cell cycle were detected by CCK8 and flow cytometry , respectively. Cell migration was exanuned by transwell assay. Finally, the expressions of transfornung growth factor-[31 ( TGF-[31 ) and vascular endothelial growth factor ( VEGF) were measured by Western blot. Results Compared with the control group , cell numbers were increased obviously in hypoxia group. Cell growth was promoted after 120 hours of incubation, the proliferation fold was 5. 00 + 0. 28 . which was 7. 64 + 0. 32 in control group (P < 0. 001 ) . The S phase was strongly accelerated . accounting for ( 10. 44 +2. 61) % ,compared with ( 20. 79 +2. 23 ) % in control group (P < 0. 05 ) . Cell migration were increased, the cell numbers was 33. 80 + 4. 20 , while which was 8. 35 + 2. 50 in control group (P < 0. 05 ) .the expressions of TGF-[31 and VEGF were enhanced significantly in hypoxia group ( P < 0. 05 ) . Hypoxia + BAMBI group : After transfection with pFLAG-BAMBI in hypoxia-induced cells ,in comparison with hypoxia group , the expression of BAMBI was up-regulated sigruficantly ; Cell numbers were decreased obviously , cell growth was inhibited after 120 hours of incubation. the proliferation fold was 5. 53 +0. 27 (P <0. 001) ,the S phase of cell cycle was blocked , accounting for ( 18. 75 +2. 92 ) % , cell migration were strongly decreased , the cell numbers was 13. 00 + 2. 80 ( P < 0. 05 ) , the expressions of TGF-[31 and VEGF were suppressed significantly in pFLAG-BAMBI transfected cells as compared with hypoxia group (P < 0. 05 ) . Conclusion The overexpression of BAMBI inhibits hypoxia induced proliferation and nugration in RF/6A cells .thus may inhibit the retinal neovascularization.

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更新日期/Last Update: 2016-05-04