缺氧对小鼠角膜血管内皮生长因子及可溶性fms-样酪氨酸激酶受体-1表达的影响

目的观察缺氧对小鼠角膜组织中血管内皮生长因子（vascular endothelial growth factor,VEGF）及可溶性fins一样酪氨酸激酶受体-1（soluble fms-like tyrosine kinase receptor-1, sFlt-1 ）表达的影响并探讨两者之间的关系。方法实验用健康雌性成年昆明小鼠70只，分为缺氧组和正常对照组，缺氧仓的氧气体积分数为8％～10％，缺氧时间分别为1、3、5、7、10、14d。用变性Western blot技术检测小鼠在缺氧条件下角膜组织中总VEGF表达的变化，用非变性Western blot技术检测小鼠在缺氧条件下角膜组织中游离及结合VEGF表达的变化，用逆转录聚合酶链反应技术检测小鼠在缺氧条件下角膜组织中VEGF、sFlt-1、缺氧诱导因子-1（hypoxia in-ducible巧玲珑factor-1，HIF-1）的基因表达变化。结果缺氧14d内未见小鼠角膜新生血管。缺氧组总VEGF及结合VEGF表达增加，随着缺氧时间的延长而增强，与正常对照组比较差异有统计学意义（P〈0.05），但缺氧14d内均未见游离VEGF表达。缺氧组VEGF、sFh-1、HIF-1的基因表达均增加，随着缺氧时间的延长而增强，与正常对照组比较差异有统计学意义（P〈0．05），其中VEGF的基因表达与HIF．1的基因表达呈正相关（r=0．993），sFlt-1的基因表达与HIF-1的基因表达呈正相关（r=0．997）。结论缺氧可以诱导小鼠角膜组织中VEGF和sFlt-1表达的增加，在缺氧的条件下，小鼠角膜组织中VEGF和sFlt-1呈平衡状态。sFlt-1可作为一种内源性VEGF拮抗因子治疗眼部新生血管性疾病。（中国眼耳鼻喉科杂志，2008，8：286-288）

Effect of hypoxia on the expression of vascular endothelial growth factor and soluble fms-like tyrosine kinase receptor-1 in mice cornea

Objective To investigate the expression of vascular endothelial growth factor（VEGF） and soluble fins-like tyrosine kinase receptor-1 （sFlt-1） in mice cornea during hypoxia and the relationship between these two factors. Methods Seventy mice were divided into two groups （hypoxia group and control group）. The hypoxia group mice were placed in a hypoxia chamber with 8% to 10% oxygen for 1, 3, 5, 7, 10, 14 days respectively. Reducing Western blot was used to examine the protein expression of total VEGF, while non-reducing Western blot was used to examine the protein expression of free and bound VEGF. Real time-polymerase chain reaction was used to examine the gene expression of VEGF, sFlt-1 and hypoxia induced factor-1（HIF-1）. Results There was no corneal neovascularization during 14 days of hypoxia. Compared with the controls, the gene expression of VEGF, sFlt-1 and HIF-1, the protein expression of total and bound VEGF all increased markedly （P〈0.05）. But there was no free VEGF protein expression during 14 days of hypoxia. The differences between the hypoxia and the control group were significant. A positive relationship was found between the gene expression of VEGF and HIF-1, sFh-1 and HIF-1. Conclusions The increased protein expression of VEGF and sFlt-1 in cornea of mice can be induced by hypoxia. VEGF and sFlt-1 came to a balance in cornea of mice on the condition of hypoxia, sFlt-1 could be an inhibitor to VEGF on ocular neovascularization. （ Chin J Ophthalmol and Otorhinolaryngo1,2008,8：286-288）