CD105 shRNA对激光诱导的脉络膜新生血管的干预作用及其机制  

The Effects of CD105 Gene Silencing to Experimental Choroidal Neovascularization in BN Rats

作　　者：屈超义 唐罗生[4] 曾杰西[4] 罗静[4] 陈百华[4] 魏为[4] 

机构地区：[1]陕西省眼科医疗中心 [2]西安市第四医院 [3]西安交通大学医学院附属广仁医院,陕西西安710004 [4]中南大学湘雅二医院眼科,湖南长沙410011

出　　处：《现代生物医学进展》2011年第22期4250-4255,共6页Progress in Modern Biomedicine

基　　金：教育部高等学校博士学科点专项科研基金(20060533026);湖南省科技厅计划项目(05FJ3057)

摘　　要：目的:观察CD105shRNA对激光诱导的大鼠脉络膜新生血管的抑制作用,并初步探讨其作用机制。方法:40只BN大鼠单眼采用半导体激光建立CNV模型。随机取20只大鼠于建模后1天使用Pgenesil-eng2转染大鼠视网膜和脉络膜作为实验组。在建模后第14天行FFA检查,观察实验组与对照组视网膜激光斑的渗漏情况。各取5只实验组和5只对照组大鼠,行脉络膜铺片,检测并比较脉络膜新生血管渗漏面积。另32只BN大鼠任取一眼建立CNV模型,其中任取20只大鼠于建模后次日进行Pgen-esil-eng2转染,作为实验组。12只建模眼作为对照组,未建模眼作为空白对照组。于基因转染后1w,2w,3w和4w各取5只实验组大鼠和3只对照组大鼠眼球,获取每个时间点实验组、对照组和空白对照组的脉络膜组织。检测各组每个时间点CD105和VEGF在mRNA水平的表达。结果:FFA显示光凝后第14天时,对照组的渗漏率为63.2%,实验组为24.6%。实验组的BN大鼠眼底渗漏点数较对照组少,渗漏强度较弱。两组间比较有显著性差异。脉络膜铺片结果显示:2周时对照组大鼠的CNV面积为(31.22±1.46)×103μm2,实验组大鼠的CNV渗漏面积为(14.46±0.82)×103μm2,两组间比较有显著性差异。RT-PCR结果显示:实验组VEGF mRNA及CD105 mRNA的表达变化规律与对照组相似,但各个时间点的表达量较对照组均明显下降,其中实验组VEGFmRNA于2w时的表达约为对照组的36.7%;实验组CD105 mRNA在第2w时约为对照组的21.68%。结论:通过沉默CD105基因的表达可以抑制大鼠CNV的生成,下调VEGF的表达可能是其作用机制之一。CD105基因有望成为CNV的基础研究热点和临床治疗的新靶点。Objective： To investigate the effect of CD 105 gene silencing on BN rat CNV model. Methods： The effective CD105 shRNA express plasmid was transinfected into BN rat CNV model by subretina injection. The result of CD105 gene silencing to CNV model was evaluated by FFA, quantitive analysis of CNV. The expression of VEGF and CD105 at mRNA level was evaluated by reverse transcription- polymerise chain reaction. Results： In CD105shRNA-treated group and CNV model control group, the incidence of fluores- cein leakage was 24.6% and 63.2%, respectively, at 14 day after photocoagulation. There was significant difference between the two groups. The result of quantitive analysis of CNV showed the area of CNV leakage is （14.46±0.82）× 10^3μm2 and （31.22 ±1.46）× 10^3μm2, respectively, in CD105 shRNA-treated group and CNV model control group, there was significant difference between the two groups. Two weeks after laser photocoagulation, CD105 and VEGF express at a high level in CNV model control group. But in CD105shRNA-treated group CD105 and VEGF expression was at a low level. Conclusion： CD105 gene silencing at early stage could in- hibit the formation of CNV in vivo. Down-regulation of VEGF expression may be one of it＇s mechanisms.CD 105 plays an important role in early stage of CNV, it is hopeful to be a target of CNV therapy.

关 键 词：脉络膜新生血管 CD105 VEGF mRNA RNAI 

分 类 号：Q95-3[生物学—动物学] R774.5[医药卫生—眼科]