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The experimental study on the expression of recombinant plasmid pEGFP-N1-IGF-1 in the injured spinal cord of rats
Author(s): 
Pages: 223-228
Year: Issue:  4
Journal: JOURNAL OF SPINAL SURGERY

Keyword:  大鼠脂质体质粒转基因胰岛素样生长因子Ⅰ脊髓损伤聚合酶链反应;
Abstract: 目的 探讨重组质粒pEGFP-N1-IGF-1在大鼠损伤脊髓内表达及mRNA定量分析,为基因治疗SCI提供依据.方法 建立改良Nystrom's大鼠脊髓不完全损伤动物模型,应用基因直接注射法,将阳离子脂质体与重组质粒pEGFP-N1-IGF-1混合后注入大鼠脊髓损伤部位,以空白质粒及生理盐水对照,转染1周后取材,行冰冻切片并于荧光显微镜下观察绿色荧光蛋白表达情况;行Real Time PCR脊髓内IGF-1的mRNA定量表达,并进行统计学分析.结果 通过阳离子脂质体Transfectam reagent成功介导重组质粒pEGFP-N1-IGF-1在大鼠脊髓内转染其神经细胞,并表达EGFP和IGF-1的融合蛋白,通过Real time PCR方法进行mRNA定量分析,进一步证实了重组质粒pEGFP-N1-IGF-1能在脊髓内明确地表达IGF-1,pEGFP-N1-IGF-1转基因组与IGF-1组和对照组mRNA表达相对数比较均有明显显著性差异(P<0.01).结论 重组质粒pEGFP-N1-IGF-1转染后能在脊髓内明确表达IGF-1.证明阳离子脂质体介导的IGF-1体内转基因方法的可行性,为今后应用IGF-1基因治疗脊髓损伤提供了理论依据.
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