原花青素对氧诱导视网膜病变新生血管形成及血管内皮生长因子表达的抑制研究  

Inhibitory effect of proanthocyanidin on neovascularization and VEGF expression in oxygen-induced retinopathy

作　　者：杨璐[1] 李冬平[1] 杨芬 杜淑芳[1] 牛红蕾 刘小燕[1] 董京艳[1] 高昭 张蕴达[1] Yang Lu;Li Dongping;Yang Fen;Du Shufang;Niu Honglei;Liu Xiaoyan;Dong Jingyan;Gao Zhao;Zhang Yunda(First Division,Department of Retinal Disorders,Shanxi Eye Hospital,Taiyuan 030002,China)

机构地区：[1]山西省眼科医院玻璃体视网膜一科,太原030002

出　　处：《中国药物与临床》2022年第2期101-105,共5页Chinese Remedies & Clinics

基　　金：山西省应用基础研究项目(面上青年基金)(201601D202093)。

摘　　要：目的探讨原花青素(PC)对C57小鼠氧诱导视网膜病变新生血管形成的抑制作用。方法采用随机数字表法将105只7日龄C57小鼠随机分为正常组、氧诱导视网膜病变(OIR)组、阴性对照组(PBS)组、低剂量PC组、中剂量PC组、高剂量PC组和Lucentis组(阳性对照组),每组15只。将实验组小鼠在高氧分压环境下饲养5 d后,于出生后第12天(P12)移出氧箱。正常组:未进氧箱饲养。各组小鼠置于的常氧空气环境中继续饲养5 d,直到出生后第17天,接下来的实验根据所设的不同的组别和时间点处死小鼠并取材。P12-P16,各实验组给予相对应的干预。P17,摘除各组小鼠眼球行视网膜铺片,进行Isolectin GS-IB4荧光染色,并用image pro plus软件分析。实时荧光定量聚合酶链反应(real-time PCR)检测各组小鼠视网膜中血管内皮生长因子(VEGF)mRNA水平。蛋白质印迹法检测各组小鼠视网膜中VEGF的蛋白表达水平。结果根据各组P17小鼠视网膜铺片免疫荧光结果显示,在OIR小鼠P12~P16注射PC,能显著减少OIR中视网膜病理性新生血管簇的形成及星形胶质细胞的损害,并呈PC剂量依赖型,高剂量PC组效果最佳。P17,小鼠视网膜VEGF的mRNA水平及蛋白表达:与正常对照组比较,OIR组、PBS组表达上调,差异有统计学意义(P<0.05),2组之间比较差异无统计学意义(P>0.05)。与OIR组比较,PC组下调与Lucentis组表达下调,差异有统计学意义(P<0.05)。高剂量PC组与Lucentis组2组之间比较差异无统计学意义(P>0.05)。结论PC以剂量依赖的方式下调氧OIR小鼠视网膜组织中VEGF的表达,从而抑制视网膜新生血管的产生。Objective To investigate the inhibitory effect of proanthocyanidin(PC)on neovascularization in C57 mice with oxygen-induced retinopathy(OIR).Methods Using a random number table method,1057-day-old C57 mice were randomly divided into the normal group,OIR group,PBS group(negative control),low-dose,medium-dose and high-dose PC groups,and the Lucentis group(positive control)(n=15 each).The mice to be modeled for OIR were fed under high oxygen partial pressure for 5 days,and were released from the oxygen chamber on day 12 postnatum(P12).The normal group was fed without use of oxygen chamber.The mice in each group were kept in a normoxic environment for 5 days until day 17 postnatum(P17).Subsequently,the mice in each group were sacrificed and harvested for tissues according to group setting and time points.On P12 through P16,each experimental group was given corresponding intervention.On P17,eyeballs were harvested from each group of mice and used for retina flat-mount preparations which were subjected to Isolectin GS-IB4 fluorescence staining and Image Pro Plus software analysis.Real-time PCR was used to examine the VEGF mRNA expression,and Western blotting was used to measure the VEGF protein expression,in retina of mice in each group.Results Immunofluorescence staining of retina flat-mount preparations in each group on P17 showed that PC injection during P12 through P16 significantly reduced the pathologic neovascularization and astrocyte damage in the retina of OIR mice in a dose-dependent manner,and such effect was the most prominent in high-dose PC group.On P17,the mRNA and protein expression levels of mice retina VEGF were up-regulated in the OIR and PBS groups compared to the normal control group with statistically significant difference(P<0.05),but there was no difference between the OIR and PBS groups(P>0.05).Compared with the OIR group,the mice retina VEGF mRNA and pro-tein expression was down-regulated in PC groups the Lucentis group,with statistically significant difference(P<0.05).There was no significant

关 键 词：原花青素 氧诱导视网膜病变 视网膜新生血管 血管内皮生长因子 

分 类 号：R774.1[医药卫生—眼科]