YKL-40在激光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 YKL-40 in choroidal neovascularization model in mice induced by laser

作　　者：宾玥[1] 刘彦尧 丁琳[1] 彭惠[1] Bin Yue;Liu Yanyao;Ding Lin;Peng Hui(Department of Ophthalmology, The First Affdiated Hospital of Chongqing Medical University;Chongqing Key Laboratory of Ophthalmolog;Chongqing Eye Institut)

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,重庆400016

出　　处：《重庆医科大学学报》2018年第6期863-868,共6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:探讨在激光诱导的小鼠脉络膜新生血管(choroidal neovascularization,CNV)模型中几丁质酶-3样蛋白-1(chitinase-3-like-1,YKL-40)的表达及意义。方法:将40只成年雄性C57BL/6J小鼠以随机数字表法按照造模后取材时间的不同随机分为光凝后7 d组和14 d组,各20只。设定每组小鼠左眼为正常对照眼,右眼通过532 nm激光光凝视网膜诱导小鼠脉络膜新生血管模型,分别于光凝后7 d和14 d心脏灌注2×106F1TC-dextran荧光素,行视网膜色素上皮(retinal pigment epithelium,RPE)/脉络膜复合体铺片以及苏木精-伊红染色法(hematoxylin-eosin staining,HE)鉴定CNV模型。采用实时荧光定量PCR法检测各组小鼠视网膜神经层(简称神经视网膜)和RPE/脉络膜复合体中YKL-40与血管内皮生长因子(vascular endothelial growth factor,VEGF)的m RNA水平变化。免疫荧光染色法观察YKL-40在各组小鼠眼组织中的表达及定位。结果:脉络膜铺片及HE染色均证实激光造模成功,光凝后7 d和14 d均有CNV形成。在光凝后7 d组中,神经视网膜中YKL-40和VEGF的m RNA相对表达量明显升高[(1.939±0.209),(4.017±1.312)],与正常组[(1.002±0.076),(1.017±0.219)]相比差异均有统计学意义(t=13.320,P=0.006;t=5.457,P=0.012),而RPE/脉络膜复合体中YKL-40和VEGF表达[(0.968±0.381),(1.192±0.354)]较正常组[(1.004±0.101),(1.021±0.240)]无明显改变(t=-4.519,P=0.053;t=2.450,P=0.134);在光凝后14d组,神经视网膜和RPE/脉络膜复合体中YKL-40和VEGF的m RNA表达水平[(3.174±1.583),(3.045±1.430),(12.669±4.512),(8.254±2.968)]与正常组[(1.013±0.173),(1.043±0.371),(1.037±0.347),(1.078±0.462)]相比均明显升高,且差异具有统计学意义(t=4.777,P=0.041;t=3.508,P=0.039;t=4.827,P=0.040;t=12.800,P=0.006)。Pearson相关性分析提示在激光造模后YKL-40的表达与VEGF具有正相关性。免疫荧光发现YKL-40主要表达于光凝后小鼠视神经节细胞层、内丛状层和内核层,且光凝后7 d和14 d YKL-40表达[(0Objective ：To explore the expression and significance of ehifinase-3-1ike- 1 （YKIL-40） in elmroidal neovascularization （CNV） models in mice. Methods：Forty adult male C.57Bl/6J mice were divided into day 7 group and day 14 group post laser coagu- lation randomly according to the sampling time, with twenty mice in each group. The left eyes of mice were set as normal control and the right eyes were established CNV models by 532 nm laser photocoagulafion. Retinal pigment epithelium（RPE）/choroidal flalmounts after cardiac perfusion with FITC-dextran and HE staining on the 7th and 14th day post laser were used to identify the CNV models. The mRNA levels of YKL-40 and vascular endothelial growth lactor（VEGF） in neumretina and RPE/chomidal complex were detected by real-time PCIL The localizalions and expressions of YKL- 40 in eyes of each group were determined by immunofluores cence staining. Results：Both RPE/choroidal flatmount and HE staining confirmed the success of model and the CNV formation on day 7 and day 14 after laser coagulation. Real-lime PCR showed that in neuroretina the expressions of YKL-40 and VEGF[（1.939± 0.209）, （4.017 ± 1.312）] were higher in day 7 group than control group[（ 1.002± 0.076）, （ 1.017 ± 0.219）],and the differences were significant （t= 13.320, P=0.O06 ; t=5.457, P=0.O 12 ）. However, in RPE/choroidal complex, no significant differences （ t= -4.519,P=0.053 ;t=2.450,P=-0.134） were observed between control group[（ 1.004 ± 0.101 ）, （1.021 ± 0.240）] and day 7 group[（0.968 ± 0.381）, （1.192 ± 0.354）]. Futhermore,the mRNA levels of YKL-40 and VEGF in day 14 group[（3.174 ± 1.583）, （3.005± 1.430）, （12.669 ± 4.512）, （8.254 ± 2.968）] were higher than those in control group[（ 1.013± 0.173）, （1.043 ± 0.371 ）, （1.037 ± 0.347）, （1.078 ± 0.462）] both in neuroretina and in RPE/choroidal complex tissues, and the differences were significant（t=4.777 ,P=0.041 ;t= 3.508,P=-0.039;t=4.827,P=0.040;t=12.80

关 键 词：YKL-40 血管内皮生长因子 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]