肝细胞导向的促红细胞生成素基因转移在肾性贫血基因治疗的初步研究

The study on in vivo erythropoietin gene transfer for treatment of renal anemia in animal model

目的探索用受体介导方法对肾性贫血基因治疗的可行性及疗效。方法构建一种克隆了促红细胞生成素（ＥＰＯ）基因的ＥＢＶ复制子载体ｐＥＰＯ，该载体与半乳糖基化组蛋白结合形成可溶性的核酸蛋白复合物，以静脉注射的方式将复合物导入通过喂饲腺嘌呤造成肾性贫血大鼠，恢复正常饮食２周后，采血样检测血红蛋白的含量和红细胞数，并与对照组进行比较。结果注射ｐＥＰＯ复合物组的红细胞数和血红蛋白含量都有明显升高，实验组和对照组的红细胞数分别为每毫升４９２个和４０７个；血红蛋白含量分别１１．４ｇ／ｍｌ和９．１ｇ／ｍｌ。两组比较，Ｐ＜０．０１。提示ＥＰＯ基因已被导入动物体内并表达出目的蛋白，但肾衰症状并不改善。结论以上结果证明通过受体介导的基因转移技术可将ＥＰＯ基因导入实验动物体内并进行表达。

To explore the possibility of gene therapy to renal anemia,the receptor mediated erythropoietin (EPO) gene transfer in vivo had been tested.EPO gene was inserted to an EBV replicon expression vector to form the plasmid pEPO, Renal anemia rat had been induced by feeding with adenine,and 450μg of the pEPO was delivered by perfusion into the rat peripheral circulation in the form of soluble DNA/galactosylate histone complex.13 days after injection,the number of peripheral red cells was counted and the concentration of hemoglobin was measured.The results demonstrated that symptom of anemia had been significantly improved(P<0.01),when compared with that of the control test.(the red cell number was 492vs 409,and the hemoglobin was 11.4vs 9.1)