G补缀FHA域血管生成因子1在糖尿病视网膜新生血管形成中的作用  

Study on the role of angiogenic factor with G patch and FHA domains 1 in diabetic retinal neovascularization

作　　者：李蓉[1] 姚国敏[1] 闫红林[2] 王莉[3] 蔡天志[3] 袁博博 巨伟 王丽娜 Rong Li;Guo-Min Yao;Hong-Lin Yan;Li-Wang;Tian-Zhi Cai;Bo-Bo Yuan;Wei Ju;Li-Na Wang(Department of Ophthalmology,the First Affiliated Hospital of Xi'an Medical University,Xi'an 710077,Shaanxi Province,China;Department of Drug Clinical Trial Institution,the First Affiliated Hospital of Xi'an Medical University,Xi'an 710077,Shaanxi Province,China;Department of Scientific Research,the First Affiliated Hospital of Xi'an Medical University,Xi'an 710077,Shaanxi Province,China;Translational Medicine Center,Xi'an Ninth Hospital Affiliated to Medical College of Xi'an Jiaotong University,Xi'an 710054,Shaanxi Province,China)

机构地区：[1]西安医学院第一附属医院眼科,中国陕西省西安市710077 [2]西安医学院第一附属医院药物临床试验机构,中国陕西省西安市710077 [3]西安医学院第一附属医院科研科,中国陕西省西安市710077 [4]西安交大医学院附属市九院转化医学中心,中国陕西省西安市710054

出　　处：《国际眼科杂志》2020年第9期1487-1492,共6页International Eye Science

基　　金：国家自然科学基金项目(No.81500726);陕西省卫生健康科研基金项目(No.2018D074);西安市科技局医学研究项目[No.201805097YX5SF31(4)];科技厅自然科学基金配套基金(No.XYFYPT-2020-07)。

摘　　要：目的:观察G补缀FHA域血管生成因子1(AGGF1)在糖尿病视网膜组织及高糖条件下视网膜血管内皮细胞中的表达,以及AGGF1对视网膜血管内皮细胞增殖、迁移和管腔形成的影响。方法:将C57BL/6J小鼠随机分为对照组和糖尿病视网膜病变(DR)模型组,采用免疫组化法检测视网膜组织中AGGF1的蛋白表达。将体外培养的恒河猴脉络膜视网膜血管内皮细胞(RF/6A细胞)随机分为对照组(低糖环境培养)和高糖组(培养基中加入25mmol/L D-葡萄糖),采用免疫荧光法检测细胞中AGGF1的蛋白表达。然后将RF/6A细胞分为对照组和AGGF1处理组,分别采用CCK-8法、Transwell法和Matrigel检测细胞增殖、迁移及管腔形成。结果:AGGF1蛋白在视网膜各层均有表达,在血管内皮细胞中也有明显表达,AGGF1在DR组视网膜中的表达(0.17±0.05)明显强于对照组(0.07±0.02)(P<0.05)。AGGF1蛋白在高糖组和对照组RF/6A细胞中均有表达,AGGF1在高糖下RF/6A细胞中的表达(0.63±0.10)明显强于对照组(0.40±0.03)(P<0.05)。处理12h,AGGF1组细胞增殖率(114.88%±0.84%)明显高于对照组(100.00%±2.17%)(P<0.05);处理24h,AGGF1组细胞增殖率(157.35%±1.89%)明显高于对照组(142.77%±0.50%)(P<0.05);处理48h,AGGF1组细胞增殖率(185.39%±1.90%)明显高于对照组(160.17%±1.33%)(P<0.05)。处理12h,AGGF1组细胞迁移数(127.00±7.00个)明显多于对照组(90.33±6.66个)(P<0.05)。处理12h,AGGF1组细胞管腔数(33.67±1.15个)明显多于对照组(15.33±3.51个)(P<0.05);AGGF1组细胞分支总长度(8226.33±288.55μm)明显多于对照组(6463.33±938.01μm)(P<0.05)。结论:糖尿病视网膜组织和高糖诱导的视网膜血管内皮细胞表达AGGF1蛋白明显增多,AGGF1可促进视网膜血管内皮细胞增殖、迁移和管腔形成,提示AGGF1可能参与了DR的视网膜新生血管形成。AIM:To observe the effect of AGGF1 on the proliferation,migration and tube formation of retinal endothelial cells in diabetic retinal tissue and high glucose conditions.METHODS:C57BL/6J mice were randomly divided into the control group and diabetic retinopathy(DR)model group.The cultured rhesus monkey choroido-retinal endothelial cells(RF/6A cells)were randomly divided into the control group(cultured in low-glucose environment)and the high-glucose group(cultured in medium with 25mmol/L D-glucose),and the AGGF1 protein expression in the cells was detected by immunofluorescence assay.RF/6A cells were then divided into the control group and AGGF1 treatment group,and cell proliferation,migration and tube formation was detected by CCK-8,Transwell and Matrigel,respectively.RESULTS:AGGF1 protein was expressed in all layers of the retinas and in vascular endothelial cells.The expression of AGGF1 in the retinas of DR group(0.17±0.05)was significantly higher than that of the control group(0.07±0.02)(P<0.05).AGGF1 protein was expressed in RF/6A cells in both the high glucose group and the control group,and the expression of AGGF1 in RF/6A cells under high glucose was significantly higher(0.63±0.10)than that in the control group(0.40±0.03)(P<0.05).After 12h of treatment,the cell proliferation rate(114.88%±0.84%)in the AGGF1 group was significantly higher than that in the control group(100.00%±2.17%)(P<0.05).After 24h of treatment,the cell proliferation rate of the AGGF1 group(157.35%±1.89%)was significantly higher than that of the control group(142.77%±0.50%)(P<0.05).After 48h of treatment,the cell proliferation rate of the AGGF1 group(185.39%±1.90%)was significantly higher than that of the control group(160.17%±1.33%)(P<0.05).After 12h of treatment,the number of migrated cells(127.00±7.00)in the AGGF1 group was significantly higher than that in the control group(90.33±6.66)(P<0.05).After 12h of treatment,the number of tube formation(33.67±1.15)in the AGGF1 group was significantly higher than that in the control

关 键 词：G补缀FHA域血管生成因子1 糖尿病视网膜病变 高糖 视网膜血管内皮细胞 血管生成 

分 类 号：R73[医药卫生—肿瘤]