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通过基因治疗的方法补充胰岛素已用于实验性治疗胰岛素依赖型糖尿病(IDDM)。本研究构建了含有重组人前胰岛素原基因的裸质粒DNA载体(pCMV-IN),将其肌肉注射人链脲佐菌素(STZ)诱发的糖尿病C57小鼠体内,并辅以电穿孔方法,以获得在体胰岛素转基因治疗。该质粒载体表达的胰岛素mRNA,可通过RT-PCR方法在转基因局部的骨骼肌组织中检测到。在接受pCMV-IN注射的糖尿病小鼠中,血浆胰岛素水平显著升高,达到了未注射STZ的正常对照小鼠的水平,且胰岛素的表达可持续至少35 d。pCMV-IN质粒注射转基因治疗显著降低了糖尿病小鼠在第7至35 d的血糖水平,其下降幅度约6 mmol/L;转基因治疗也显著降低了严重糖尿病小鼠的死亡率,其第6周时的死亡率由100％降为37％。结果表明,直接肌肉注射含入前胰岛素原基因裸质粒可获得胰岛素的有效表达,显著降低糖尿病小鼠的血糖水平并降低严重糖尿病小鼠的死亡率。裸质粒注射胰岛素转基因治疗有望成为IDDM的一种有效治疗手段。
Insulin supplementation by gene therapy has been used to experimentally treat insulin-dependent diabetes mellitus (IDDM). In this study, we constructed a naked plasmid DNA vector (pCMV-IN) containing recombinant human proinsulin gene and intramuscular injection of streptozotocin (STZ) -induced diabetic C57 mice supplemented with electroporation Obtain in vivo insulin transgenic therapy. The mRNA expression of this plasmid vector can be detected by RT-PCR in transgenic skeletal muscle tissue. In diabetic mice receiving pCMV-IN injection, plasma insulin levels increased significantly, reaching the level of normal control mice not injected with STZ, and insulin expression persisted for at least 35 days. Transfection with plasmid pCMV-IN resulted in a significant decrease of blood glucose level in diabetic mice from day 7 to day 35 with a decrease of about 6 mmol / L. Transgenic treatment also significantly reduced the mortality of severe diabetic mice. The sixth week When the mortality rate from 100% to 37%. The results showed that the direct intramuscular injection of naked plasmid containing proinsulin gene can be obtained effective expression of insulin, significantly reduce the blood glucose levels in diabetic mice and reduce the mortality of severe diabetic mice. Naked plasmid injection of insulin gene therapy is expected to become an effective treatment for IDDM.