中文摘要：

目的：构建胰岛素样生长因子1的真核细胞表达质粒，分析pcDNA3．1-hIGF-1体内转基因的町行性。方法：实验于2004—06／09在吉林大学基础医学院动物实验室完成。①选取清洁级成年雄性Wistar大鼠12只，随机数字表法分为胰岛素样生长因子1组、模型对照组、正常对照组，4只／组。②聚合酶链法从人的肝细胞cDNA文库中克隆出胰岛素样生长因子1cDNA，然后定向插入真核细胞表达载体pcDNA3．1中，酶切电泳和基因测序鉴定插入质粒的人源性胰岛素样生长因子基因序列。③胰岛素样生长因子1组应用直接注射法将阳离子脂质体和pcDNA3．1-hIGF1混合后转染至大鼠胸段脊髓组织中；模型对照组注射等剂量pcDNA3．1和阳离子脂质体混合液。正常对照组未作任何处理。1周后应用免疫组化法观察人源性胰岛素样生长因子在大鼠脊髓中的表达。结果：实验选取清洁级成年雄性Wistar大鼠12只，全部进入结果分析。①聚合酶链反应片段鉴定结果：琼脂糖电泳显示，扩增带大小与预期的人源性胰岛素样生长因子cDNA相同。②重组的真核表达质粒的鉴定结果：酶切鉴定显示，重组的真核细胞表达质粒pcDNA3．1-hOGF-1所含的IGF—1cDNA序列和插入方向均正确。③基因转染后各组胰岛素样生长因子1免疫组化测定结果：基因转染1周后，胰岛素样生长因子1组脊髓组织内阳性细胞数明显高于模型对照组和正常对照组（48．2±5.3,29．3±4．2，23．8±8．3，P〈0．01）。结论：阳离子脂质体介导pcDNA3．1-hIGF1体内转基因的方法是可行的。

英文摘要：

AIM： To construct eukaryotic expression plsmid of human insulin-like growth factor 1 （hIGF-1） and identify the feasibility of pcDNA3.1-hIGF-1 transfection in vivo. METHODS： The experiment was conducted in the animal experimental laboratory of Basic Medicine Department, Jilin University from June to September 2004. ①Twelve clean grade adult male rats were randomly divided into IGF-1 group, model group and control group with 4 rats in each group. ②hIGF-1cDNA was cloned from human liver cDNA library by PCR and inserted into pcDNA3.1, eukaryotic expression vector, to form recombinant plasmid, pcDNA3.1-hIGF-1, fragment electrophoresis and gene sequence measurement were undertaken to ensure the inserted gene. ③In IGF-1 group, mixture of pcDNA3.1-hIGF-1 and Iiposome was transfected into rat spinal cord with direct injection method： the model group was injected with the same amount mixture of pcDNA3.1 and liposome; the control group was not given any intervention. One week later, immunohistochemistry was adopted to detect the expression of IGF-1 in the rat spinal cord. RESULTS： All the 12 rats were involved in the result analysis. ①Results of PCR： The agarose electrophoresis showed that the size of amplified belt was accorded with hlGF-1cDNA as anticipated.②Results of recombinant plasmid： The digested method showed that the IGF-1cDNA sequence and insert direction in the pcDNA3.1-hIGF-1 were all correct. ③ Immunohistochemistry results of hIGF-1： After 1 week of transfection, the number of positive cells in spinal tissue of the IGF-1 group was obviously higher than the model group and control group （48.2±5.3, 29.3±4.2, 23.8 ±8.3, P 〈 0.01）. CONCLUSION： Transfection of pcDNA3.1-hIGF-1 into spinal cord by liposome is feasible.