脯氨酸羟化酶2对氧诱导视网膜病变新生血管生成的影响  

Effect of proline hydroxylases 2 on neonatal mice by oxygen-induced retinopathy

作　　者：屈晓杰[1] 王雨生[1] 张鹏[1] 储昭节[1] 

机构地区：[1]第四军医大学西京医院眼科全军眼科研究所,陕西省西安市710032

出　　处：《眼科新进展》2011年第6期511-515,共5页Recent Advances in Ophthalmology

基　　金：国家自然科学基金资助(编号:30672291;30872818);德国洪堡基金会(Alexander von Humboldt Foundation)仪器设备捐赠基金(V-8151/02085;To YS Wang)资助~~

摘　　要：目的观察脯氨酸羟化酶2(proline hydroxylase2,PHD2)对氧诱导小鼠视网膜病变动物模型新生血管生成的影响。方法 40只7d龄的C57BL/6J幼鼠随机分为4组,即正常对照组、高氧组、300mg.kg-1和500mg.kg-1PHD2抑制剂处理组,每组各10只。后3组放入动物氧舱,在氧气含量为体积分数(75±2)%的高氧环境中连续饲养5d,正常对照组于正常大气环境中饲养。300mg.kg-1和500mg.kg-1PHD2抑制剂处理组在结束高浓度给氧后,腹腔注射不同剂量的PHD2抑制剂(黄芩素),高氧组和正常对照组给予腹腔注射生理盐水,每天1次,连续5d。于末次注射结束后处死小鼠,Western blotting法检测视网膜中PHD2蛋白的表达,视网膜铺片法观察视网膜血管的情况,HE染色后计数突破内界膜的内皮细胞核数,免疫组织化学法检测视网膜a-SMA和VEGF蛋白的表达。结果 Western blotting显示视网膜中存在PHD2蛋白的表达,且高氧组蛋白表达量明显高于正常对照组(P=0.002),正常对照组高于300mg.kg-1、500mg.kg-1PHD2抑制剂组(P=0.004、0.001)。视网膜铺片结果显示高氧组自视盘发出的血管扩张迂曲,中周部视网膜有大量新生血管生成,近视盘周围有明显的血管无灌注区;300mg.kg-1和500mg.kg-1PHD2抑制剂处理组较高氧组明显改善。突破视网膜内界膜的内皮细胞核数正常对照组、高氧组、300mg.kg-1和500mg.kg-1PHD2抑制剂处理组分别为(0.60±0.84)个、(39.40±5.17)个、(15.80±1.93)个和(6.30±1.57)个,4组比较差异有统计学意义(P<0.05),高氧组最多,正常对照组最少。免疫组织化学染色结果显示300mg.kg-1和500mg.kg-1PHD2抑制剂处理组a-SMA表达较高氧组明显增加;高氧组、300mg.kg-1及500mg.kg-1PHD2抑制剂处理组的VEGF表达量均较正常对照组明显升高,300mg.kg-1和500mg.kg-1PHD2抑制剂处理组与高氧组比较,VEGF表达量无明显降低。结论视网膜存在PHD2的表达,抑制其表达后能有效改善高氧所致小鼠视网膜血管的Objective To investigate effects of proline hydroxylase 2（PHD2）on neovascularization of oxygen-induced retinopathy（OIR）models.MethodsForty C57BL/6J mice at postnatal 7 days were randomly divided into 4 groups：normal control group,hyperoxia group,300 mg·kg-1 PHD2 inhibitor treatment group and 500 mg·kg-1 PHD2 inhibitor treatment group,10 mice in each group.The normal control group lived in normal circumstances and another three groups were exposed to（75±2）% oxygen for 5 days and then transferred to the normal air.After the end of the hyperoxia,300 mg·kg-1 PHD2 inhibitor treatment group and 500 mg·kg-1 PHD2 inhibitor treatment group were given different doses of inhibitor at 1 time per day,and for 5 days.Hyperoxia group and normal control group were only given normal saline injection.Mice in each group were executed at the last injection.Western blotting was used to detect PHD2 protein expression in retina,and retinal preparation was used to observe condition of retinal vessels.HE staining was used to count endothelial cell nucleus breaking through the internal limiting membrane.Retinal a-SMA and VEGF protein were detected by immunohistochemical staining.ResultsThe PHD2 protein expressed in retina by Western blotting,and higher in hyperoxia group than that in normal control group（P=0.002）,higher in normal control group than that in 300 mg·kg-1 and 500 mg·kg-1 PHD2 inhibitor treatment groups（P=0.004,0.001）.Retinal preparation showed that blood vessels expanded and were with tortuous from in hyperoxia group,lots of neovascularization in retina at central perimeter,and obvious blood no perfusion area was found around near optic disc.300 mg·kg-1 PHD2 inhibitor treatment group and 500 mg·kg-1 PHD2 inhibitor treatment group had better improvement than hyperoxia group.The endothelial cell nucleus breaking through the internal limiting membrane in the normal control group,hyperoxia group,300 mg·kg-1 PHD2 inhibitor treatment group and 500 mg·kg-1 PHD2 inhibitor treatment group were 0.60±0.8

关 键 词：视网膜新生血管 脯氨酸羟化酶2 氧诱导视网膜病变 鼠 

分 类 号：R774.1[医药卫生—眼科]