[1]李蓉,杜军辉,姚杨,等.血管内皮细胞生长因子抑制剂对视网膜血管内皮细胞自噬的促进作用[J].眼科新进展,2018,38(10):935-939.[doi:10.13389/j.cnki.rao.2018.0221]
 LI Rong,DU Jun-Hui,YAO Yang,et al.The promoting effect of vascular endothelial growth factor inhibitor on autophagy of retinal vascular endothelial cells[J].Recent Advances in Ophthalmology,2018,38(10):935-939.[doi:10.13389/j.cnki.rao.2018.0221]
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血管内皮细胞生长因子抑制剂对视网膜血管内皮细胞自噬的促进作用/HTML
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《眼科新进展》[ISSN:1003-5141/CN:41-1105/R]

卷:
38卷
期数:
2018年10期
页码:
935-939
栏目:
实验研究
出版日期:
2018-10-05

文章信息/Info

Title:
The promoting effect of vascular endothelial growth factor inhibitor on autophagy of retinal vascular endothelial cells
作者:
李蓉杜军辉姚杨李晓艳
710077 陕西省西安市,西安医学院第一附属医院眼科,西安医学院全科医学院五官科教研室,西安医学院医院管理研究所(李蓉,李晓艳),西安医学院第一附属医院中心实验室(姚杨);710054 陕西省西安市,西安交通大学附属西安市第九医院眼科(杜军辉)
Author(s):
LI RongDU Jun-HuiYAO YangLI Xiao-Yan
Department of Ophthalmology,Teaching and Researching Office of Ophthalmology and Otorhinolaryngology,Hospital Management Institute of Xi’an Medical University (LI Rong,LI Xiao-Yan),Central Laboratory (YAO Yang),Xi’an 710077,Shaanxi Province,China;Department of Ophthalmology,Xi’an Ninth Hospital Affiliated to Medical College of Xi’an Jiaotong University (DU Jun-Hui),Xi’an 710054,Shaanxi Province,China
关键词:
自噬血管内皮细胞生长因子缺氧视网膜血管内皮细胞
Keywords:
autophagyvascular endothelial growth factorhypoxiaretinal vascular endothelial cell
分类号:
R774.1
DOI:
10.13389/j.cnki.rao.2018.0221
文献标志码:
A
摘要:
目的 观察血管内皮细胞生长因子(vascular endothelial growth factor,VEGF)抑制剂对小鼠视网膜血管内皮细胞(retinal vascular endothelial cells,RVECs)自噬水平的影响。方法 将RVECs随机分为五组:正常对照组、缺氧对照组、自噬抑制剂3-甲基腺嘌呤(3-methyladenine,3-MA)组、VEGF抑制剂(anti-VEGF)组和3-MA+anti-VEGF组。采用Western blot法检测RVECs中两种自噬标志蛋白微管相关蛋白1 轻链 3 (LC3)及Beclin-1的表达;计算绿色荧光蛋白(green fluorescent protein,GFP)阳性斑点细胞占比;透射电子显微镜下观察细胞内形成的自噬体的超微结构。结果 正常对照组、缺氧对照组、3-MA组、anti-VEGF及3-MA+anti-VEGF组的LC3-Ⅱ/LC3-Ⅰ分别为0.182±0.125、0.587±0.101、0.309±0.151、0.914±0.037及0.585±0.098;Beclin-1分别为0.205±0.035、0.590±0.120、0.425±0.082、0.842±0.087及0.607±0.022;GFP阳性斑点细胞比例分别为17.107%±3.521%、90.278%±2.684%、82.591%±4.490%、94.798%±1.760%及89.472%±3.764%。与正常对照组相比,缺氧对照组LC3-Ⅱ/LC3-Ⅰ和Beclin-1蛋白表达增高,GFP阳性斑点细胞的比例增加(均为P<0.05),同时自噬体增多。与缺氧对照组相比,3-MA组的LC3-Ⅱ/LC3-Ⅰ、Beclin-1蛋白表达水平及GFP阳性斑点细胞比率降低(均为P<0.05),同时自噬体减少;anti-VEGF组的LC3-Ⅱ/LC3-Ⅰ、Beclin-1蛋白表达水平及GFP阳性斑点细胞比率升高(均为P<0.05),同时自噬体增多。与anti-VEGF组相比,3-MA+anti-VEGF组的LC3-Ⅱ/LC3-Ⅰ、Beclin-1蛋白表达水平及GFP阳性斑点细胞比率降低(均为P<0.05),同时自噬体减少。结论 VEGF抑制剂对缺氧状态下视网膜血管内皮细胞的自噬水平有促进作用。
Abstract:
Objective To explore the effects of vascular endothelial growth factor (VEGF) inhibitor on autophagy level of mouse retinal vascular endothelial cells (RVECs).Methods RVECs were randomly divided into five groups:the normal control group,hypoxia control group,autophagy inhibition group [adding autophagy inhibitor 3-methyladenine (3-MA)],anti-VEGF group (adding VEGF inhibitor) and 3-MA+anti-VEGF group.The two autophagy marker proteins microtubule associated protein 1 light chain 3 (LC3) and Beclin-1 were examined by Western blot.The ratio of green fluorescent protein-positive (GFP+) puncta in RVECs was counted.The ultrastructure of autophagosomes formed in cells was observed under transmission electron microscopy (TEM).Results The expression ratio of LC3-Ⅱ to LC3-Ⅰ was 0.182±0.125,0.587±0.101,0.309±0.151,0.914±0.037 and 0.585±0.098,respectively,while the expression level of Beclin-1 was 0.205±0.035,0.590±0.120,0.425±0.082,0.842±0.087 and 0.607±0.022 for the normal control group,hypoxia control group,3-MA group,anti-VEGF group and 3-MA+anti-VEGF group,respectively.GFP(+) cell rate was 17.107%±3.521%,90.278%±2.684%,82.591%±4.490%,94.798%±1.760% and 89.472%±3.764%,respectively.Compared with the normal control group,the expression of LC3-Ⅱ/LC3-Ⅰ and Beclin-1 proteins in the hypoxia control group enhansed,the GFP+ cell rate increased (all P<0.05) and the formation of autophagosomes increased.Compared with the hypoxia control group,LC3-Ⅱ/LC3-Ⅰ,Beclin-1 and GFP+ cell rate in the 3-MA group decreased (all P<0.05),and the formation of autophagosomes decreased.Compared with the hypoxia control group,LC3-Ⅱ/LC3-Ⅰ,Beclin-1 and GFP+ cell rate in the anti-VEGF group increased (all P<0.05),and the formation of autophagosomes increased.Compared with the anti-VEGF group,LC3-Ⅱ/LC3-Ⅰ,Beclin-1 and GFP+ cell rate in the 3-MA+anti-VEGF group decreased (all P<0.05),and the formation of autophagosomes decreased.Conclusion VEGF inhibitor has a promoting effect on the autophagy level of retinal vascular endothelial cells under hypoxia.

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备注/Memo

备注/Memo:
国家自然科学基金(编号:81500726);西安市科技局医学研究项目[编号:201805097YX5SF31(4)];陕西省普通高校青年杰出人才支持计划
更新日期/Last Update: 2018-09-28