虫草素对高糖培养的恒河猴脉络膜-视网膜血管内皮细胞血管新生的影响  

Anti-angiogenic mechanism of cordycepin on rhesus macaque choroid-retinal endothelial cell line cultured in high glucose condition

作　　者：朱小丽[1] 孙小凤[1] 赖铭莹[1] 

机构地区：[1]深圳市眼科医院暨南大学附属深圳市眼科医院深圳眼科学重点实验室,中国广东省深圳市518000

出　　处：《国际眼科杂志》2016年第7期1237-1241,共5页International Eye Science

基　　金：深圳市知识创新计划基础研究项目(No.JCYJ20130402145545766);深圳市科技计划项目(No.201202144)~~

摘　　要：目的:观察虫草素对高糖培养的恒河猴脉络膜-视网膜血管内皮细胞(RF/6A)血管新生作用的影响,探讨其可能的作用机制。方法:培养RF/6A细胞,分为正常对照组(NC组)、高糖对照组(HG组)、高糖加不同浓度的虫草素组(HG+10μg/m L组、HG+50μg/m L组、HG+100μg/m L组)。采用CCK8法检测各组细胞的增殖活性;采用Transwell实验检测各组细胞的迁移;采用Matrigel检测各组管腔形成的情况;采用Western-blot检测各组细胞中VEGF和VEGFR2蛋白表达的情况。结果:与NC组相比,HG组RF/6A的增殖活性增加(P<0.05)。不同浓度的虫草素对RF/6A的增殖均有抑制作用,随着虫草素浓度增加,细胞活性下降。HG+10μg/m L组、HG+50μg/m L组、HG+100μg/m L组的增殖活性抑制率分别为(10.2±0.9)%、(23.4±1.5)%、(31.1±1.2)%,与HG组比较差异均有统计学意义(P<0.05)。NC组、HG组、HG+10μg/m L组、HG+50μg/m L组、HG+100μg/m L组细胞迁移个数分别为55.6±2.70、87.4±2.40、65.4±2.7、57.8±2.38、62.4±2.77个。与NC组相比,HG组迁移细胞数量增加(P<0.05);不同浓度虫草素组细胞迁移数量随着虫草素浓度的增加而减少,与HG组相比,差异具有统计学意义(P<0.05)。NC组、HG组、HG+10μg/m L组、HG+50μg/m L组、HG+100μg/m L组的细胞管腔形成个数分别为18.7±2.08、25.7±1.52、19.9±1.57、16.3±2.51、5.7±1.72个。与NC组相比,HG组细胞管腔形成个数增加(P<0.05);不同浓度虫草素组细胞管腔形成个数随着虫草素浓度的增加而减少,与HG组相比,差异具有统计学意义(P<0.05)。与NC组相比,HG组VEGF和VEGFR2蛋白表达的量增加(P<0.05);不同浓度虫草素组中细胞内VEGF和VEGFR2蛋白表达的量均低于高糖对照组(P<0.05)。结论:虫草素可能通过抑制高糖下VEGF和VEGFR2蛋白的表达,抑制RF/6A细胞增殖、迁移和管腔形成,进而抑制血管新生。AIM： To investigate the angiogenesis effect and protective mechanism of cordycepin on rhesus macaque choroid- retinal endothelial （ RF/ 6A） cell line cultured in high glucose condition. METHODS： Cultured RF/ 6A cells were divided into normal control group, high glucose group and high glucose （HG） ＋ different concentration cordycepin groups （HG＋ 10μ g/ mL group, HG＋ 50μ g/ mL group, HG＋ 100μ g/mL group）. The cell proliferation was assessed using cholecystokinin octapeptide dye after treated for 48h. The cell migration was investigated by a Transwell assay. The tube formation was measured on Matrigel. Furthermore, the impact of cordycepin on high glucose - induced＆amp;nbsp;activation of VEGF and VEGF receptor 2 （VEGFR-2） was tested by Western blot analysis. RESULTS： Compared with normal control group, cell viability markedly increased in high glucose group （ P 〈0. 05）. Cordycepin inhibited RF/ 6A cell proliferation in a dose- dependent fashion： 10. 2 ± 0. 9%, 23. 4 ± 1. 5% and 31. 1±1. 2% inhibition as the concentrations of cordycepin were 10, 50 and 100μ g/ mL, respectively. The difference had statistically significant （P〈0. 05） compared with high glucose group. The number of cell migration were 55. 6±2. 70, 87. 4 ± 2. 40, 65. 4 ± 2. 7, 57. 8 ± 2. 38, 62. 4 ± 2. 77 in normal control group, high glucose group and HG＋10μ g/mL group, HG ＋ 50μ g/ mL group, HG ＋ 100μ g/ mL group respectively. Migration of RF/ 6A conspicuously increased in high glucose group （ P 〈 0. 05） compared with normal control group; while showing a gradually reducing trend with the increase of cordycepin dose and a statistically significant difference compared with high glucose group （P〈0. 05）. The number of tube formation were 18. 7±2. 08, 25. 7 ± 1. 52, 19. 9 ± 1. 57, 16. 3 ± 2. 51, 5. 67 ± 1. 72 in the abovementioned group. Similarly showing a gradually reducing trend with the increase of cordycepin dose and a statistically significant difference with high glu

关 键 词：虫草素 高糖培养 恒河猴脉络膜-视网膜血管内皮细胞 血管内皮生长因子 血管新生 

分 类 号：R276.7[医药卫生—中医五官科学]