重组CFB-siRNA在脉络膜新生血管中抑制作用的实验研究  

Inhibition efficacy of CFB-siRNA on laser-induced choroidal neovascularization in rat

作　　者：李凡[1] 尚庆丽[1] 郝玉华[1] 马景学[1] 叶存喜[1] 王鑫[1] 

机构地区：[1]河北医科大学第二医院眼科,河北石家庄050000

出　　处：《河北医科大学学报》2017年第11期1305-1309,1365,共6页Journal of Hebei Medical University

基　　金：河北省应用基础研究计划重点基础研究项目(09966111D)

摘　　要：目的补体旁路途径在脉络膜新生血管(choroidal neovascularization,CNV)的发生发展中起重要作用,补体因子B(complement factor B,CFB)作为补体旁路途径的重要因子可成为阻断补体活化的靶点。本研究探讨重组CFB-siRNA在实验性CNV中的抑制作用与机制。方法取Brown Norway大鼠45只随机分为空白对照组、实验对照组、实验组各15只(30只眼)。空白对照组不给予任何干预措施,实验对照组与实验组均激光光凝建立大鼠CNV模型。实验对照组在CFB表达高峰前日尾静脉注射生理盐水0.5μL,实验组同一时间尾静脉注射CFB-SiRNA(0.5μL/75μg),均隔日注射1次,共注射3次。各组分别于光凝后3、7、14、21、28d进行荧光素眼底血管造影(fluorescein fundusangiography,FFA),根据荧光素渗漏程度对各光凝斑评分并检测CNV的生长情况;采用免疫组织化学法检测各组视网膜脉络膜组织中CFB、血管内皮生长因子(vascular endothelial growth factor,VEGF)、碱性成纤维细胞因子(basic fibroblast growth factor,BFGF)的表达情况,并测定其灰度值。结果 FFA显示:实验组与实验对照组比较,在光凝后7、14、21、28dCNV发生率差异有统计学意义(P<0.05)。免疫组织化学检测结果显示:空白对照组正常大鼠CFB、VEGF和BFGF在视网膜和脉络膜中的表达非常弱。CFB表达情况:光凝后3d实验组与实验对照组CFB表达达最高峰,随后表达减少,光凝后7d仍有少量表达,光凝后14~28d表达趋于稳定。实验对照组VEGF、BFGF在光凝后7~14d表达明显增多,光凝后21d达高峰;实验组VEGF、BFGF在光凝后14、21、28d表达减少。3组CFB、VEGF和BFGF灰度值在组间、时点间、组间·时点间交互作用差异均有统计学意义(P<0.05)。结论尾静脉注射CFB-siRNA可抑制实验性CNV的发展,通过抑制CFB,阻断补体旁路途径,减少CNV形成过程中VEGF、BFGF的表达。Objective Alternative complement pathway plays important roles in the pathogenesis and development of choroidal neovascularization(CNV).Complement factor B(CFB),an essential factor for the alternative complement pathway,has been considered as a target to block the activity of complement.In this study,we aim to investigate the inhibitive effects and the potential mechanism of recombinant CFB-siRNA in the CNV.Methods Fortyfive Brown Norway rats were randomly divided into:blank control(n=15),subject to no treatment;control group(n= 15), which was subject to CNV induction by laser photocoagulation,followed by administration of normal saline(0.5μL)via caudal vein injection one day before the expression peak of CFB,and test group,which was subject to laser photocoagulation to induce CNV,followed by administration of CFB-siRNA(0.5μL/75μg)via caudal vein injection one day before the expression peak of CFB.The injection of normal saline and CFB-siRNA was performed every two days for thrice.Fundus fluorescein angiography(FFA)was performed on day 3,7,14,21,and 28 after laser photocoagulation.Photocoagulation score was determined according to the leakage of the fluorescein,based on which to detect the growth of CNV.Expression of CFB, VEGF,and BFGF in the retina and the choroid was measured using immunohistochemical method.Results FFA showed that the difference of CNV incidence between the experimental group and the experimental group was statistically significant at 7,14,21 and 28 days after photocoagulation(P<0.05).In the blank control,the expression of CFB,VEGF,and BFGF in the retina and choroid was comparatively lower as revealed by immunohistochemical method.For the expression of CFB,peak value was obtained 3 days after photocoagulation in the experimental and control group,followed by down-regulation of CFB.Low content of CFB was still detected 7 days after photocoagulation,while the expression was stable from day 14 to day 28.In the control group,significant up-regulation was observed in the BEGF and BFGF 7-14 days

关 键 词：脉络膜新生血管化 补体因子B 血管内皮生长因子 

分 类 号：R773.4[医药卫生—眼科]