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目的探讨阳离子聚合物包装IL-1ra重组质粒、角膜原位转染治疗角膜碱烧伤的疗效。设计实验性研究。研究对象角膜碱烧伤大鼠动物模型。方法制作Wistar大鼠角膜碱烧伤模型,随机分为2组,Ⅰ组为阴性对照(12只),结膜下注射生理盐水。Ⅱ组为IL-1ra基因治疗组(18只),以PcDNA3.1质粒作为IL-1ra基因表达载体,阳离子聚合物为转染介质,角膜基质显微注射20μg IL-1ra质粒。碱烧伤后不同时间点(3、7、21天)处死实验动物取下角膜,HE染色分析角膜组织病理变化,比较两组角膜透明性和新生血管生长情况,对角膜内浸润的炎性细胞进行计数。主要指标角膜透明性和新生血管情况,角膜组织病理变化及炎性细胞计数。结果碱烧伤造模后,对照组角膜浸润水肿明显,且随时间延长而加重,角膜基质内大量炎性细胞浸润和粗大新生血管形成。IL-1ra基因治疗组的角膜基质轻、中度水肿,角膜中性粒细胞、淋巴细胞等炎性细胞计数在不同时间点均少于对照组,角膜新生血管管径均较细。结论角膜基因原位转染可使IL-1ra在局部有效表达,抑制碱烧伤引起的角膜免疫炎症反应,为眼碱烧伤的治疗提供了新的选择。
Objective To investigate the effect of cationic polymer-encapsulated IL-1ra recombinant plasmid and corneal in situ transfection on corneal alkali burn. Design experimental research. Objective To investigate the animal model of corneal alkali burns in rats. Methods Wistar rat models of corneal alkali burn were randomly divided into two groups. Group Ⅰ was negative control (n = 12) and saline was injected subconjunctivally. In group Ⅱ, IL-1ra gene therapy group (18), pcDNA3.1 plasmid as IL-1ra gene expression vector, cationic polymer as transfection medium, microinjection of 20μg IL-1ra plasmid into corneal stroma. At different time points after alkali burn (3, 7 and 21 days), the cornea were sacrificed and corneal histopathological changes were observed by HE staining. Corneal transparency and neovascularization were compared between the two groups, and infiltration of corneal inflammatory cells count. The main indicators of corneal transparency and neovascularization, pathological changes of corneal tissue and inflammatory cell count. Results After alkali burn, the corneal infiltration and edema in the control group were obvious, and aggravated with time. A large number of inflammatory cell infiltration and thick neovascularization were formed in corneal stroma. Corneal stromal light, moderate edema, corneal neutrophil, lymphocyte and other inflammatory cell counts in IL-1ra gene therapy group were less than those in control group at different time points, corneal neovascularization diameter was smaller. Conclusion In situ transfection of corneal gene can effectively express IL-1ra locally and inhibit corneal immunoinflammatory reaction induced by alkali burn, which provides a new choice for the treatment of ocular alkali burn.