环氧合酶-2抑制剂Rofecoxib抑制小鼠视网膜新生血管形成的实验研究  

Inhibitive effects of COX-2 inhibition Rofecoxib on retinal neovascularization in rats

作　　者：刘宁宁[1] 柳力敏[1] 万超[1] 胡悦东[1] 周赟[1] 才娜[1] 陈蕾[1] 

机构地区：[1]中国医科大学附属第一医院眼科,辽宁省沈阳市110001

出　　处：《眼科新进展》2010年第7期629-632,共4页Recent Advances in Ophthalmology

基　　金：辽宁省自然科学基金资助(编号:20082119)~~

摘　　要：目的探讨环氧合酶-2(cyclooxygenase-2,COX-2)抑制剂Rofecoxib抑制小鼠视网膜新生血管形成的机制。方法同日出生7d的2窝C57BL/6小鼠各10只,随机分为治疗组及未治疗组。2组在高氧环境下饲养5d后,置于正常环境中饲养。治疗组每天1次腹腔注射COX-2抑制剂Rofecoxib(15mg.kg-1),未治疗组注入等量的生理盐水,连续5d。生后第17天摘取小鼠的眼球用于检查。荧光素血管灌注观察视网膜血管形态,HE染色计数与内界膜有关系的血管内皮细胞核。免疫组织化学检测视网膜中COX-2和VEGF蛋白表达;RT-PCR检测COX-2和VEGFmRNA表达。结果 2组小鼠每个切面均可见突破内界膜的血管内皮细胞核,未治疗组平均每个切面为(22.56±2.13)个,治疗组小鼠平均每个切面为(5.39±1.52)个,2组比较,差异有显著统计学意义(P<0.001)。未治疗组:COX-2和VEGF蛋白表达在内核层、神经节细胞层和突破视网膜内界膜的新生血管。治疗组:外核层、内核层、神经节细胞层未见COX-2蛋白阳性表达;VEGF蛋白在外核层、内核层无阳性表达,神经节细胞层个别细胞浆内可见微弱表达。COX-2的光密度(opticaldensity,OD)值未治疗组为3.64±0.75,治疗组为1.02±0.55,2组比较,差异有显著统计学意义(P<0.001);VEGF的OD值未治疗组为4.10±0.51,治疗组为3.74±0.49,2组比较,差异也有显著统计学意义(P<0.001)。治疗组视网膜中COX-2吸光度值为1.67±0.38,未治疗组视网膜中COX-2吸光度值为4.45±0.24。治疗组鼠视网膜COX-2mRNA基因表达比未治疗组降低了62%(P<0.05)。治疗组视网膜中VEGF吸光度值为3.17±0.21,未治疗组鼠视网膜VEGF吸光度值为7.74±0.15。治疗组鼠视网膜VEGFmRNA基因表达比未治疗组降低了59%(P<0.05)。结论 COX-2抑制剂Rofecoxib通过抑制VEGF和COX-2蛋白及mRNA的表达来抑制小鼠视网膜新生血管形成。Objective To investigate the inhibitive effects of cyclooxygenase-2 （COX-2） inhibition Rofecoxib on retinal neovascularization in rats.Methods Two postnatal 7 days broods of C57BL/6 rats born in the same day （10 cases in each brood） were divided into treated group and untreated group.The rats of two groups were breed in hyperxia of 5 days,then breed in the normal condition.The rats in treated group were injected the COX-2 inhibition Rofecoxib （15 mg·kg-1）,once a day;The rats in untreated group were injected the equal physiological saline of 5 days.At the postnatal 17 days,the eyeballs were extracted.The fluorescein perfusion method was adopted to assess the vascular pattern,HE staining methods was applied to count the nuclei of new vessels related to the inner retinal,immunohistochemistry was used to detect the expression of COX-2 and VEGF protein,and RT-PCR for COX-2 mRNA and VEGF mRNA was performed.Results The nucleus of endothelial cell breaking through the inner limiting membrane could be found in all slices of two groups,the number of the nucleus in each slice were （22.56±2.13） cases in the treated group and （5.39±1.52） cases,the difference was significant compared between two groups（P0.001）.In the untreated group,the expression of COX-2 and VEGF protein were showed in the inner nuclear layer,the ganglion cell layer and the neovascularization breaking through the inner limiting membrane;In the treated group,there was no expression of COX-2 protein in the outer nuclear layer,inner nuclear layer and ganglion cell layer,and no expression of VEGF protein in the outer nuclear layer and inner nuclear layer,little in the endochylema of some ganglion cells.The optical density of COX-2 and VEGF were 3.64±0.75 and 4.10±0.51 in the untreated group,1.02±0.55 and 3.74±0.49 in the treated group,the difference were all significant compared between two groups （all P0.001）.The absorbance value of COX-2 and VEGF were 1.67±0.38 and 3.17±0.21 in the treated group,4.45±0.24 and 7.74±0.15 in

关 键 词：视网膜新生血管 环氧合酶-2 ROFECOXIB 

分 类 号：R774.1[医药卫生—眼科] Q959.836[医药卫生—临床医学]