RT-PCR测定大鼠脉络膜新生血管色素上皮衍生因子研究

Detect PEDF mRNA by RT-PCR in Experimental Choroidal Neovascularization Induced by Krypton Laser

目的建立氪激光诱导的BN大鼠脉络膜新生血管(choroidal neovascularization,CNV)模型,建立RT-PCR方法检测大鼠视网膜及相关组织色素上皮衍生因子(pigment epithelium-derived factor,PEDF)mRNA性。研究PEDF mRNA与CNV生长的关系,从而探讨在CNV生长中的作用。方法对6组24只BN大鼠应用氪激光(波长647nm)实验眼视网膜进行光凝,功率360mW、光斑直径50μm、曝光时间0.05s,围绕视盘等距光凝8个点,创建CNV模型。采用半定量反转录聚合酶链反应(RT-PCR)方法检测PEDF mRNA在大鼠视网膜中的表达水平的变化。结果FFA光凝斑盘状荧光素渗漏,证实CNV形成。视网膜中PEDF mRNA表达水平呈先升高后降低的变化。结论氪激光诱导BN大鼠产生CNV模型,病理结构稳定,RT-PCR方法适合做大鼠动物模型PEDF mRNA的半定量检测,PEDF的变化与新生血管的发送更有关。

Objective This study was designed to establish an experimental Choroidal neovascularization （CNV） model in BN rats, which could exhibit high reproducibility and to evaluate the expression of pigment epithelium - derived factor （ PEDF）. Methods 30 rats were received krypton laser. 1 days, 3 days, 7 days, 14days, 21 days and 28 days after photocoagnlation, fundus fluorescein angiography （FFA） wAS performed. Total BNA were abstracted from retinal and choroidal tissue and the mBNA of PEDF was detected by BT - PCB. Results FFA showed flurescein leakage from 1 week to 4 weeks after photocoagulation. The mBNA of PEDF were in significantly difference during 4 weeks by BT - PCB. Conclusions Kryton laser photocoagulation can be successfully used to produced CNV experimental model in the BN rat. This study gave us a effective way to evaluate the mechanism of CNV by BT - PCR.