重组人内皮抑素抑制角膜新生血管的实验研究

目的研究重组人内皮抑素（rHES）对碱烧伤诱导大鼠角膜新生血管（CNV）的抑制效应。方法60只Wistar大鼠按随机数字表法分为正常组、阳性对照组、rHES组3组，每组各20只。阳性对照组右眼角膜碱烧伤后不治疗；rHES组右眼角膜碱烧伤后立即100mg／mL rHES点眼，每日3次；20只正常鼠不做任何干预处理作为正常组。观察碱烧伤后1、4、7、10、14d各组角膜情况及CNV的生长面积。碱烧伤后第16天处死动物，取角膜行组织病理学检查。免疫组织化学、PCR检测血管内皮生长因子（VEGF）蛋白量和mRNA在角膜组织的表达，透射电镜下检查碱烧伤后角膜组织的超微结构变化。结果rHES组角膜在碱烧伤后各时间点新生血管面积均小于阳性对照组（t4d=3．294，P=0．040；t7d=5．391，P=0．000；t10d=6．560，P=0．000；t14d=10．346，P=0．000）。角膜碱烧伤后第16天各组VEGF蛋白表达量的差异有统计学意义（F=337．62，P=0．00），各组VEGF mRNA表达量的差异有统计学意义（F=114．43，P=0．00）。透射电镜检查显示正常组结构正常，rHES组较阳性对照组角膜结构变化小。PCR结果发现rHES组VEGF mRNA的相对表达量低于阳性对照组，但高于正常组。结论rHES用于眼表可有效抑制碱烧伤诱导的CNV。

Inhibition of recombinant human endostatin on corneal neovascularization

Objective Recombinant human endostatin （rilES） is proved to inhibit neovascularization （NV）. Present study was to determine the anti-angiogenesis action of rilES on alkali-burn-induced corneal neovascularization （CNV） in rats. Methods Corneal alkali-burn model was established by putting the filter paper soaking 1 mol/L NaOH in the central cornea for 40 seconds in the right eyes of 40 healthy Wistar rats and then were randomly divided into model group and experimental group and 20 rats for each. The 100 mg/mL of rilES drops was topically administered in the experimental group after corneal alkali burning 3 times per day for 15 days. Other 20 normal rats worked as controls. The area of CNV was culculated quantitatively on day 1,4,7,10,14 after establishment of model. The rats were sacrificed and corneal samples were obtained for the uhrastructure examination under the transmission electron microscope. Expression of vascular endothelial growth factor （VEGF） protein was detected using immunohistochemistry. Expression of VEGF mRNA was determined by RT-PCR. Use of animals followed the Association for Research in Vision and Ophthalmology. Results The area of corneal neovascularization in rilES group was significantly smaller than that in model group at various time points （t4d = 3. 294, P = 0. 040;t7 d = 5. 391, P = 0. 000;t10d = 6. 560, P = 0. 000 ; t14 d = 10. 346, P = 0. 000）. The infiltration of inflammatory cells in stroma and severe corneal edema were seen in model group,and the inflammatory reaction was mild in rilES group under the light microscope. The less ultrastrueture change of cornea was found in rilES group compared with model group under the transmission electron microscope in 16 days after corneal alkali burning. The expression of VEGF protein in cornea tissue was significantly different among normal, model and rilES group （F = 337.62, P = 0. 00） with the lower expression level in rilES group. The expression level of VEGF mRNA in cornea was significantly different among normal, m