欢迎访问《现代生物医学进展》编辑部！

目的：了解内皮抑素（ES）转基因治疗子宫内膜异位症大鼠模型的疗效。方法：构建子宫内膜异位症大鼠模型，选择建模成功的大鼠为实验研究对象，随机分为ES 转染组（I 组）24 只、载体对照组（II 组）20 只和阴性对照组（III 组）20 只。I 组病灶局部注射 lipofectamine-endo-pBud 复合物进行基因转染，II 组注射lipofectamine-pBud 复合物，III 组注射PBS 用于对照。通过实时荧光定量 PCR 法检测异位病灶中ES 基因的相对表达量，用Western-blot 测定ES-HA 融合蛋白及ES 蛋白的相对表达量，来判断转染成功与否。用ELISA 法对大鼠血清中ES 及血管内皮细胞生长因子（VEGF）水平进行测定，用免疫组化SP 法对ES、基质金属蛋白酶 -2（MMP-2）以及微血管密度（MVD）的表达进行测定，用游标卡尺对转染前后各组大鼠异位病灶的长、宽进行测量，计算体积，分析各指标实验前后的差异，观察内皮抑素转基因治疗子宫内膜异位症大鼠模型的疗效。结果：注射相应试剂后2 周，I 组异位病灶组织中ES 基因的相对表达量高于两对照组（P<0.05），有ES-HA 融合蛋白表达，且ES 蛋白的相对表达量显著高于两对照组（P<0.01）；I 组血清中ES 水平显著高于两对照组（P<0.01），VEGF 水平显著低于两对照组（P<0.01），三组ES 与VEGF 在血清中的表达水平呈负相关（r=-0.805）；I 组异位病灶组织中ES 表达明显高于两对照组（P<0.01）；MMP-2 的表达明显少于两对照组（P<0. 01）；MVD 明显少于两对照组（P<0.01）；三组ES 与MMP-2 在异位内膜中的表达呈负相关（r=-0.700）；I 组异位病灶体积明显小于两对照组（P<0.01）。结论：阳离子脂质体LipofectamineTM2000 介导的重组质粒endo-pBud 病灶内直接注射法可以成功实现ES 在子宫内膜异位症大鼠异位内膜中的表达，并对子宫内膜异位症大鼠模型有治疗作用。

英文摘要:

Objective: To observe the curative effect by transferring endostatin gene into endometriosis rat model. Methods: To establish endometriosis model in Lewis rats by deploying autotransplantation, choosing the sixty-four successful rat models as empirical study objects, divided them randomly into three groups: Injected 2 μl lipofectamine-endo-pBud compounds to I group, 2μl lipofectamine- pBud compounds to II group and 2μl PBS to III group. Taked the ectopic focus tissues to measure the opposing expression of endostatin gene by real-time fluorescent quantitative PCR, measure the opposing expression of endostatin protein and expression of ES-HA fusion protein by Western-blot. Detect the level of endostatin and VEGF in serum by ELISA. Detected the endostatin, MMP-2 and MVD in ectopic focus by immunohistochemical assay, and measured the volume of the ectopia focus by vernier cursor. Results: The opposing expressions of endostatin gene were difference between I group and the two control groups has statistical significance (P<0.05); ES-HA fusion protein can be detected in the ectopic focus in I group; The opposing expression of endostatin protein is much higher in the ectopic focus of I group than in the two control groups (P<0.01). The level of ES is much higher in I group than in the two control groups (P<0.01); The level of VEGF is much lower in I group than in the two control groups(P<0.01); There was an negative correlation between ES and VEGF in serum (r=-0.805). The expression of ES is much higher in I group than in the two control groups (P<0.01); The expression of MMP-2 is much lower in I group than in the two control groups (P<0.01); and there was an negative correlation between the expression of ES and MMP-2 in ectopia focus (r=-0.700) . The MVD is much lower in I group than in the two control groups(P<0.01); The volume of the ectopia focus is smaller in I group than in the two control groups (P<0.01) two weeks after transferring. Conclusions: ES gene and protein can expressed in endometriosis rats successfully by injecting lipofectinmine-endo-pBud compounds to the ectopic focus directly, and it has definite curative effect in endometriosis rat model.

View Full Text View/Add Comment Download reader