大鼠角膜碱烧伤后新生血管基因治疗作用的研究  

Experimental study on interleukin-1 receptor antagonist gene therapy for cautery-induced rat corneal neovascularization

作　　者：滕旭[1] 孙旭光[1] 王智群[1] 

机构地区：[1]北京市眼科研究所

出　　处：《眼科研究》2002年第5期416-418,共3页Chinese Ophthalmic Research

基　　金：北京市自然科学基金支持项目 (7992 0 1 2 )

摘　　要：目的　观察白细胞介素 1受体拮抗剂 (IL 1ra)基因对大鼠角膜碱烧伤后新生血管 (CNV)的治疗作用。方法　建立角膜碱烧伤后CNV模型 ,以pcDI质粒作为IL 1ra基因的表达载体 ,通过结膜下注射进行基因治疗。Wistar大鼠随机分为 5组 ,其中 3组为不同剂量基因质粒治疗组 ,以及地塞米松组和生理盐水对照组。利用裂隙灯显微镜观察和角膜灌注铺片法观察记录 (CNV)的生长情况 ;利用半定量逆转录聚合酶链式反应 (RT PCR)检测角膜内血管内皮生长因子(VEGF)mRNA的表达。结果　基因质粒治疗组和激素组CNV的生长均受到明显抑制。与阴性对照组相比 ,在烧伤后第 7、14、2 8天 ,5 0 μg质粒基因治疗组CNV面积抑制率分别为 34 7%、37 5 %、43 7%,差异有显著性 (P <0 0 1) ,而不同剂量基因治疗组间差异无显著性 (P >0 0 5 )。烧伤后 2 4h ,VEGFmRNA水平上升至高峰 ,为正常角膜的 5 30倍 ;第 4天为正常角膜的 2倍 ;第 7天恢复至正常。 5 0 μg质粒基因治疗组VEGFmRNA的表达水平受到明显抑制 ,2 4h时下降了48 7%(P <0 0 1) ;烧伤后 4天 ,下降了 2 7 2 %(P <0 0 1)。结论　pcDI hIL 1ra基因质粒对CNV具有抑制作用 ,其机制与抑制VEGFmRNA的上调有关。ObjectiveTo evaluate the gene therapy of interleukin-1 receptor antagonist(IL-1ra)for cautery-induced rat corneal neovascularization(CNV).MethodsIn Wistar rats,the model of CNV was set up.Rats were divided into 5 groups.Three groups received subconjunctival injection of pcDI-hIL-1ra plasmid at 25 μg, 50 μg, 100 μg respectively after cautery.Dexamethasone 0.1 mg and 0.9% of normal saline were used as positive and negative controls.Corneal angiogenesis was evaluated by the perfusing method and with slip lamp microscope.Total RNA was extracted from whole corneas at different time after cautery,and vascular endothelial growth factor (VEGF)mRNA level was determined by reverse transcription-polymerase chain reaction(RT-PCR).ResultsIn treatment groups, both neovascularization and VEGF transcription were inhibited.50 μg of pcDI-hIL-1ra inhibited 34.7% of CNV at 7 d,37.5%,at 14 d,and 43.7% at 28 d.All treatment groups had statistically significant difference compared with negative control(P<0.01).NO significant differences were found between treatment groups. VEGF 165 mRNA peaked at 24 h after cautery,a 5.3-fold increase by 24 h and a 1-fold increase by 4 d,and decreased to near baseline by d 7 after cautery.50 μg of pcDI-hIL-1ra plasmid inhibited 48.7% of VEGF upregulation at 24 h and 27.2% at 4 d(P<0.01).ConclusionspcDI-hIL-1ra plasmid inhibits the cautery-induced CNV,and the effect is associated with the inhibition of VEGF mRNA upregulation.

关 键 词：角膜碱烧伤 白细胞介素-1受体拮抗剂 角膜新生血管 基因治疗 血管内皮生长因子 动物模型 

分 类 号：R779.1[医药卫生—眼科]