药物和质粒导入途径对提高人胰岛素基因在糖尿病大鼠体内表达效能的研究

Research about expression efficacy of Human Insulin Gene in type 1 diabetes rat enhanced by medicine and the way plasmids perfused

目的通过探索药物和质粒导入途径对胰岛素基因表达效能的影响,以期提高以逆转录病毒为载体进行糖尿病基因治疗的效能,为该方法治疗糖尿病的临床实施缩短距离。方法建立链脲佐菌素诱导的1型糖尿病大鼠模型,通过是否给予HGF、T3和注入重组质粒部位的不同对含有目的基因的重组质粒的转染和整合效率进行干预,观察血糖的控制情况和胰岛素的表达水平,最后提取人鼠的肝脏作RT-PCR,判断目的基因有无表达。结果通过腹腔注入含有目的基因的重组质粒同时给予HGF、T3组的效果最佳,通过尾静脉注入含有目的基因的重组质粒同时给予HGF、T3组效果较佳,仅注入重组质粒组有轻度改善,仅注入HGF、T3组和糖尿病组无任何改善。RT-PCR检测目的基因有表达。结论在动物实验中,应用HGF、T3和通过腹腔注入可明显提高含有目的基因的重组质粒的转染和整合效率,通过胰岛素、血糖的水平和体重的变化来体现。本研究为下一步的临床实验奠定了基础。

Objective To enhance the efficacy of gene therapy for diabetes mellitus and remove the obstacles for the clinical application, this experiment explores different medicines and different sites the plasmids perfused into. Methods Type 1 diabetes mellitus model of rat is extablished by STZ. After that, efficiency of transfer and integration of the goal gene were intervened the through injecting HGF and Ts and different sites the plasmids perfused into, and later the values level of blood glucose and plasma insulin was observed. The rat＇s liver was extracted and detected the gene expresses by RT- PCR. Results The group treated with the reorganization plasmids including goal gene which were perfused into the abdominal cavity and HGF and T3 injected 24 hours ago had the best effect. The group given HGF and T3 and reorganization plasmids including goal gene which were poured into tail vein had the better effect. The group treated with only the reorganization plasmids including goal gene which were perfused into the abdominal cavity had slight effect. The group given only HGF and T3 and the group with no treatment had not any effect. There was expression of goal gene by RT-PCR. Concision In animal experiment, perfusing reorganization plasmids including goal gene into the abdominal cavity with injecting HGF and T3 24 hours ago can obviously enhance the transfer and integration efficiency. This is shown by the level of insulin and glucose and the changes of weight. So this experiment lays the foundation for the clinical experiment.