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  

Inhibitive effects of BAMBI overexpression on retinal vascular endothelial cell proliferation and migration induced by hypoxia in Rhesus monkey

作　　者：朱江[1] 许治国[1] 任梅[1] 党晓洁 

机构地区：[1]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,陕西省西安市710004

出　　处：《眼科新进展》2016年第5期418-422,共5页Recent Advances in Ophthalmology

摘　　要：目的探讨过表达激活素膜结合抑制剂(bmp and activin membrane-bound inhibitor,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+pF LAG组和缺氧+BAMBI组。Western blot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,Western blot检测转化生长因子β1(transforming growth factor-β1,TGF-β1)和血管内皮生长因子(vascular endothelial growth factor,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120 h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pF LAG-BAMBI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120 h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。Objective To explore the effects of bmp and activin membrane-bound inhibitor（ BAMBI） overexpression on retinal vascular endothelial cell（ RF /6 A） proliferation and m igration induced by hypoxia in rhesus m onkey. Meth od s Cell hypoxia m odel was established and cells were divided into four groups： Control group,hypoxia group,hypoxia ＋ p FLAG group and hypoxia ＋ BAMBI group. The expression of BAMBI was m easured by W estern blot. Cell m orphology and density were observed under inverted m icroscope. Cell proliferation and cell cycle were detected by CCK8 and flow cytom-etry,respectively. Cell m igration was exam ined by transwell assay. Finally,the expressions of transform ing growth factor-β1（ TGF-β1） and vascular endothelial growth factor（ VEGF） were measured by Western blot. Results Compared with the control group,cell num bers were increased obviously in hypoxia group. Cell growth was prom oted after1 2 0 hours of incubation,the proliferation fold was 5. 0 0 ± 0. 2 8,which was 7. 6 4 ± 0.3 2 in control group（ P 〈0. 0 0 1）. The S phase was strongly accelerated,accounting for（ 10. 44 ± 2. 61） %,compared with（ 20. 79 ± 2. 23） % in control group（ P 〈0.0 5）. Cell m igration were increased,the cell num bers was 3 3. 8 0 ± 4. 2 0,while which was 8. 3 5 ± 2. 5 0 in control group（ P 〈0. 0 5）,the expressions of TGF-β1 and VEGF were enhanced significantly in hypoxia group（ P 〈0. 0 5）. H ypoxia ＋ BAMBI group：After transfection with p FLAG-BAMBI in hypoxia-induced cells,in com parison with hypoxia group,the expression of BAMBI was up-regulated significantly; Cell num bers were decreased obviously,cell growth was inhibited after 1 2 0 hours of incubation,the proliferation fold was 5. 5 3 ± 0. 2 7（ P 〈0. 0 0 1）,the S phase of cell cycle was blocked,accounting for（ 1 8. 7 5± 2. 9 2） %,cell m igration were strongly decreased,the cell num bers was 1 3. 0 0 ± 2. 8 0（ P 〈0. 0 5）,the expressions of TGF-β1 and VEGF were suppressed significantly

关 键 词：视网膜血管内皮细胞 缺氧 BAMBI 增殖 迁移 

分 类 号：R285.5[医药卫生—中药学]