CAJ | 학술논문

目的探讨重组B因子(CFB)小干扰RNA(siRNA)抑制激光诱导的大鼠脉络膜新生血管(CNV)的效果.方法采用基因重组的方法获得重组CFB-siRNA,取棕色挪威(BN)大鼠42只84只眼,用激光光凝方式建立CNV模型,随机分为尾静脉注射组、玻璃体腔注射组、视网膜下腔注射组、对照组.3种不同的注射方式组分别设有相应的空质粒注射组.除对照组外,其余各组于激光光凝后第1、3、5天分别进行注射CFB-siRNA,尾静脉、玻璃体腔、视网膜下腔注射组的注射剂量分别为50、20、10 μg;对照组激光光凝后不给于任何干预.激光光凝前和光凝后14 d分别行荧光素眼底血管造影(FFA),根据荧光渗漏程度对各激光光斑评分来检测CNV的生成情况;免疫组织化学法检测各组大鼠脉络膜内血管内皮生长因子(VEGF)、第Ⅷ因子的表达情况.结果各组激光光斑荧光渗漏程度评分结果显示,CFB-siRNA尾静脉注射组与对照组相比荧光渗漏程度明显减轻(X~2=15.1620,P<0.05).免疫组织化学结果显示,CFB-siRNA尾静脉注射组VEGF、第Ⅷ因子的阳性表达强度在激光光凝后不同时间点之间差异无统计学意义(F=20.35,18.33;P>0.05),而与同时间点其他各组相比,明显降低(F=77.96,55.68;P<0.05).其他各组VEGF、第Ⅷ因子的阳性表达强度在激光光凝后14 d均显著强于激光光凝后7 d(F=60.89,61.12;P<0.05).结论通过下调VEGF及第Ⅷ因子的表达水平,重组CFB-siRNA能有效的抑制CNV的生成.
목적 탐토중조B인자(CFB)소간우RNA(siRNA)억제격광유도적대서맥락막신생혈관(CNV)적효과.방법 채용기인중조적방법획득중조CFB-siRNA,취종색나위(BN)대서42지84지안,용격광광응방식건립CNV모형,수궤분위미정맥주사조、파리체강주사조、시망막하강주사조、대조조.3충불동적주사방식조분별설유상응적공질립주사조.제대조조외,기여각조우격광광응후제1、3、5천분별진행주사CFB-siRNA,미정맥、파리체강、시망막하강주사조적주사제량분별위50、20、10 μg;대조조격광광응후불급우임하간예.격광광응전화광응후14 d분별행형광소안저혈관조영(FFA),근거형광삼루정도대각격광광반평분래검측CNV적생성정황;면역조직화학법검측각조대서맥락막내혈관내피생장인자(VEGF)、제Ⅷ인자적표체정황.결과 각조격광광반형광삼루정도평분결과현시,CFB-siRNA미정맥주사조여대조조상비형광삼루정도명현감경(X~2=15.1620,P<0.05).면역조직화학결과현시,CFB-siRNA미정맥주사조VEGF、제Ⅷ인자적양성표체강도재격광광응후불동시간점지간차이무통계학의의(F=20.35,18.33;P>0.05),이여동시간점기타각조상비,명현강저(F=77.96,55.68;P<0.05).기타각조VEGF、제Ⅷ인자적양성표체강도재격광광응후14 d균현저강우격광광응후7 d(F=60.89,61.12;P<0.05).결론 통과하조VEGF급제Ⅷ인자적표체수평,중조CFB-siRNA능유효적억제CNV적생성.
Objective To observe the inhibitory effects of construction of complement factor (CFB)small interference RNA (siRNA) on choroidal neovascularization(CNV)induced by photocoagulation in rats.Methods We constructed an expression vector of CFB-siRNA by cutting CFB-siRNA and plasmid pRNATU6.1/Neo.Experimental CNV was induced by photocoagulation in 42 Brown Norway rats.After the model was set up,the rats were randomly divided into tail intravenous injection,vitreous injection,subretinal injection,and control group;each group except the control one had a corresponding blank plasmid control group.CFB-siRNA was injected 1,3,and 5 days respectively after photocoagulation in the injection groups;the dosage was 50,20,and 10 μg in tail intravenous injection,vitreous injection,and subretinal injection group respectively,while no injection was give to the control group after photocoagulation.Before and 14 days after photocoagulation,fundus fluorescein angiograoph (FFA) was performed and CNV development was judged by the leakage;the expression of vascular endothelial growth factor (VEGF) and factor Ⅷ were detected by immunohistochemistry.Results The leakage of fluorescein was obvious lower in tail intravenous injection group than that in the control group (X~2=15.1620,P<0.05).The expression of VEGF and factor Ⅷ in tail intravenous injection group at different time points after photocoagulation didn't differ much (F=20.35,18.33;P>0.05);while was apparently lower than that in the other groups at different time points (F=77.96,55.68;P<0.05).All of the groups,except tail intravenous injection group,had higher expression of VEGF and factor Ⅷ 14 days after photocoagulation compared with that 7 days after photocoagulation (F=60.89,61.12;P<0.05).Conclusions Constructed CFB-siRNA can inhibite CNV by down-regulating the expression of VEGF and factor Ⅷ.