Notch-Dll4在激光诱导的大鼠脉络膜新生血管生成中的作用  

Effect of Notch-Dll4 in laser-induced choroidal neovascularization in rats

作　　者：马可[1] 李晓[1] 王雨生[1] 叶子[1] 蔡岩[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》2010年第5期401-405,410,共6页Recent Advances in Ophthalmology

基　　金：国家自然科学基金资助(编号:30872818)~~

摘　　要：目的探讨Notch信号通路中Delta样配体4(Delta-like ligand4,Dll4)在激光诱导的大鼠脉络膜新生血管(choroidal neovascularization,CNV)生成中的作用机制。方法将雄性棕色挪威(brown-Norway,BN)大鼠分为正常组、光凝组、实验组和对照组。光凝组、实验组、对照组利用532nm激光诱导建立双眼CNV模型。实验组在建立CNV模型后立刻玻璃体内注射25g.L-1Avastin10μL,对照组注射0.01mmol.L-1PBS10μL。采用免疫荧光染色检测Dll4和血管内皮生长因子(vascular endo-thelial growth factor,VEGF)在正常组和光凝组光凝后7d中的表达;采用Western-blotting和RT-PCR检测正常组和光凝组(光凝后1d、3d、7d、14d)实验组、对照组(注射后7d)Dll4和VEGF的蛋白和mRNA表达;通过HE染色和视网膜色素上皮-脉络膜-巩膜铺片检测实验组和对照组(注射后14d)CNV生成的厚度和面积。结果Dll4和VEGF在CNV区域有共表达。Dll4和VEGF蛋白及其mRNA的表达趋势一致,均在光凝后1d表达开始上升,7d至高峰,14d开始下降。玻璃体内注射Avastin后,VEGF和Dll4蛋白及其mRNA表达均下降;实验组CNV面积(24202±2608)μm2较对照组(37226±3208)μm2明显减小,差异有统计学意义(P<0.05);实验组CNV厚度(53.68±5.89)μm较对照组(82.86±7.46)μm明显减小,差异有统计学意义(P<0.05)。结论Dll4在CNV生成中具有一定作用,可能参与了VEGF调控CNV生成的过程。Objective To investigate the effective mechanism of Delta-like ligand 4（Dll4）in Notch signal pathway in choroidal neovascularization（CNV）.Methods Brown-Norway（BN）rats were randomly divided into four groups,including normal group,photocoagulation group,treatment group and control group.The experimental CNV in double eyes was induced by 532 nm laser photocoagulation in rats of photocoagulation group,treatment group and control group.As soon as photocoagulation,the rats in treatment group were treated by intravitreous injection with 10 μL Avastin（25 g·L-1）and control group were treated by intravitreous injection with 10 μL PBS（0.01 mmol·L-1）.The coexpression of Dll4 and vascular endothelial growth factor（VEGF）was detected by triple immunofluorescence in normal group,photocoagulation group at 7 days after photocoagulation.The protein and mRNA expressions of Dll4 and VEGF were examined by Western-blotting and RT-PCR in normal group,photocoagulation group at 1 day,3 days,7 days and 14 days,treatment group and control group（7 days after injection）.The thickness and the area of CNV were qualified by HE staining and choroidal flat mount in treatment group and control group（14 days after injection）.Results The coexpression of Dll4 and VEGF existed in the area of CNV.The expression of Dll4 and VEGF were with one accord,increased at 1 day after photocoagulation,peaked at 7 days after photocoagulation and then decreased at 14 days after photocoagulation.After the intravitreous injection with Avastin,the expressions of Dll4 and VEGF were all significantly decreased.The area of CNV in treatment group（24 202±2 608）μm2 were significantly decreased compared to control group（37 226±3 208）μm2,there was statistical difference（P〈0.05）.The thickness of CNV in treatment group（53.68±5.89）μm were also significantly decreased compared to control group（82.86±7.46）μm,there was statistical difference（P〈0.05）.Conclusion Dll4 plays an important role in the pathogenesis of C

关 键 词：Delta样配体4 血管内皮生长因子 脉络膜新生血管 

分 类 号：R773.4[医药卫生—眼科]