CD146在聚乙二醇诱导的小鼠脉络膜新生血管模型中的表达及意义  

Expression and clinical significance of CD146 in polyethylene glycol-induced choroidal neovascularization in a mouse model

作　　者：刘彦尧 宾玥[1] 丁琳[1] 麦雨欣 王星[1] 彭惠[1] Liu Yanyao;Bin Yue;Ding Lin;Mai Yuxin;Wang Xing(Department of Ophthalmology,The First Affiliated Hospital of Chongqing Medical University,Chongqing Key Laboratory of Ophthalmology,Chongqing Eye Institute)

机构地区：[1]重庆医科大学附属第一医院眼科、眼科学重庆市重点实验室重庆市眼科研究所

出　　处：《重庆医科大学学报》2019年第11期1492-1499,共8页Journal of Chongqing Medical University

基　　金：国家自然科学基金面上资助项目（编号：81670881）

摘　　要：目的:探讨在聚乙二醇(polyethylene glycol,PEG)诱导的小鼠脉络膜新生血管(choroidal neovascularization,CNV)模型中CD146的表达及意义。方法:将60只8周龄雄性C57BL/6J小鼠采用随机数字表法,将小鼠随机分为第5、10和15天组,每组各20只。设定每组小鼠左眼为正常对照眼,右眼为实验眼,采用在视网膜下注射PEG诱导形成脉络膜新生血管模型。造模后摘取各组小鼠眼球,制作视网膜组织切片及HE染色,鉴定CNV模型。通过比较各组视网膜HE染色切片外核层(outer nuclear layer,ONL)厚度,观察PEG的视网膜毒性作用。采用实时荧光定量PCR法检测小鼠神经视网膜和RPE/脉络膜复合体中CD146、血管内皮生长因子(vascular endothelial growth factor,VEGF)、血管内皮生长因子受体2(vascular endothelial growth factor receptor 2,VEGFR2)的mRNA水平变化。免疫组化染色法检测小鼠眼内CD146、VEGF和VEGFR2的表达。结果:HE染色和ONL层厚度比较均证实,视网膜下注射PEG造模成功且模型可靠,视网膜下注射后第5和10天均有CNV形成。实验组小鼠神经视网膜和脉络膜中CD146、VEGF、VEGFR2的mRNA表达水平与对照组相比明显升高,差异有统计学意义(F=30.412,P=0.000;F=84.974,P=0.000;F=117.423,P=0.000;F=918.786,P=0.000;F=319.110,P=0.000;F=113.896,P=0.000)。Person相关性分析提示在视网膜下注射PEG后,小鼠RPE/脉络膜复合体中CD146与VEGF和VEGFR2的表达量呈正相关(r=0.940,P=0.000;r=0.940,P=0.000;r=0.769,P=0.045;r=0.910,P=0.003;r=0.910,P=0.003;r=0.777,P=0.042)。免疫组化染色的结果显示,在造模第10天后,造模组与正常对照组相比较CD146、VEGFR2在神经节细胞层、内核层、外从状层、外核层的阳性表达均有不同程度增强(P=0.000,P=0.000,P=0.000)。结论:CD146伴随着CNV的形成表达上调,且与VEGF的表达量和VEGFR2的表达量呈正相关性,由此推断CD146可能在CNV形成的病理过程中起到至关重要的作用。Objective:To explore the expression of CD146 in polyethylene glycol(PEG)-induced choroidal neovascularization(CNV) in a mouse model and its significance.Methods:Sixty 8-week-old male C57 BL/6 J mice were randomly and equally divided into 5-day group,10-day group,and 15-day group using a random number table.The left eye was set as a normal control eye,while the right eye was set as an experimental eye.The CNV model was established by subretinal injection of PEG.After model establishment,the eyeballs of mice were collected in each group.The sections of the retina were stained with hematoxylin-eosin(HE)to evaluate the CNV model.Thickness of the outer nuclear layer(ONL)in HE-stained sections of the retina was compared between the three groups to evaluate the retinal toxicity of PEG.The mRNA levels of CD146,vascular endothelial growth factor(VEGF),and vascular endothelial growth factor receptor 2(VEGFR2)in the neuroretina and RPE/choroid complex were determined by RT-PCR.Immunohistochemical staining was used to measure the expression of CD146,VEGF,and VEGFR2 in the eyes of mice.Results:Both HE staining and thickness of the ONL confirmed that model establishment by subretinal injection of PEG was successful and reliable.The development of CNV was observed at 5 and 10 days after subretinal injection.The mRNA expression of CD146,VEGF,and VEGFR2 in the neuroretina and choroid was significantly higher in the experiment group than in the control group(F=30.412,P=0.000;F=84.974,P=0.000;F=117.423,P=0.000;F=918.786,P=0.000;F=319.110,P=0.000;F=113.896,P=0.000).The Pearson correlation analysis showed that the expression of CD146 was positively correlated with the expression of VEGF and VEGFR2 in the RPE/choroid complex after subretinal injection of PEG(r=0.940,P=0.000;r=0.940,P=0.000;r=0.769,P=0.045;r=0.910,P=0.003;r=0.910,P=0.003;r=0.777,P=0.042).The results of immunohistochemical staining showed that at 10 days after model establishment,the model group had significantly higher expression of CD146 and VEGFR2 in the ganglion cel

关 键 词：CD146 血管内皮生长因子 血管内皮生长因子受体2 脉络膜新生血管 年龄相关性黄斑变性 

分 类 号：R774[医药卫生—眼科]