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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 explore the effect of drug and plasmid import on insulin gene expression in order to improve the efficacy of retrovirus-mediated gene therapy for diabetes and to shorten the clinical implementation of this method for the treatment of diabetes. Methods Streptozotocin-induced type 1 diabetic rat models were established. The effects of transfection and integration of recombinant plasmids containing the gene of interest on HGF, T3 and the site of recombinant plasmids injected were observed to observe the control of blood glucose And insulin expression levels, and finally the human liver was extracted for RT-PCR to determine whether the expression of the target gene. Results The recombinant plasmids containing target gene were injected intraperitoneally simultaneously with HGF. T3 group had the best effect. The recombinant plasmids containing the gene of interest were injected into the caudal vein simultaneously with HGF. The results of T3 group were better than those of the control group , Only HGF injection, T3 group and no improvement in diabetic group. The target gene was detected by RT-PCR. Conclusion In animal experiments, the transfection and integration efficacies of recombinant plasmids containing the gene of interest were significantly enhanced by the application of HGF, T3 and intraperitoneal injection, as indicated by changes in insulin, blood glucose levels and body weight. This study laid the foundation for the next clinical trial.