腺病毒载体介导血管抑素防治兔高危角膜移植术后新生血管化的实验研究

目的探讨腺病毒载体介导血管抑素靶向基因治疗兔高危角膜移植术后新生血管化的作用及其可能机制。方法 建立30只兔高危角膜新生血管移植受体模型,同时进行含有血管抑素基因的腺病毒载体的构建,行双眼角膜移植术,术中采用角膜原位转染法用微量进样器行角膜基质注射,右眼为实验眼注入病毒液,左眼为对照眼注入生理盐水。术后不同时间比较角膜植片透明度、水肿和新生血管程度,HE染色观察新生血管生长情况和炎症反应情况;免疫组化检测AS、VEGF在角膜植片中的表达;透射电镜观察超微结构。结果⑴实验组角膜新生血管术后较对照组显著减少(P<0.05)。⑵实验组比对照组炎症浸润程度轻。⑶.术后实验组AS的表达水平均显著高于对照组(P<0.05)。⑷术后实验组VEGF的表达水平均显著低于对照组(P<0.05)。结论腺病毒载体介导血管抑素可有效防治兔高危角膜移植术后新生血管化。

Effects of recombinant adenovirus-mediated angiostatin on inhibiting high-risk keratoplasty neovascularization of rabbit

Objective To investigate the expressions of recombinant Adenovirus-mediated angiostatin on inhibiting high-risk keratoplasty neovascularization of rabbit and evaluate the effect and possible mechanism.Methods 30 high-risk corneal neovascularization rabbit model were established and simultaneously angiostatin gene adenovirus vector was build.Double corneal transplant surgery were operated.Situ transfection method using micro-injector was taken to corneal injection.The right eye were injected by the virus solution for the experimental eye,left eye for the control eyes were used with saline.After corneal transparency degree of edema and neovascularization was compared.HE staining of the growth of CNV and inflammatory reactions;immunohistochemistry AS,VEGF expression in corneal film and TEM ultrastructure were observed.Results ⑴ Corneal neovascularization after experimental group was significantly reduced than the control group（P ＆lt;0.05）.⑵ The experimental group was mild inflammatory infiltration than the control group.⑶ AS expression after the experimental group were significantly higher（P ＆lt;0.05）.⑷ VEGF expression after the experimental group were significantly lower than the control group（P ＆lt;0.05）.Conclusion Recombinant adenovirus-mediated angiostatin can inhibit high-risk keratoplasty neovascularization of rabbit effectively.