辐照对水流动力学注射介导的小鼠体内基因表达的影响

Effects of Irradiation on Hydrodynamic Injection-mediated Gene Expression in Mice

目的探寻辐照对水流动力学注射(HGT)介导的荧光素酶报告基因在小鼠体内的表达影响,从而评估水流动力学注射方法在小鼠造血干细胞移植模型中应用的有效性。方法将实验鼠分为辐照组和非辐照组,尾静脉大体积快速注射带有荧光素酶报告基因的真核表达质粒PGL3-luc。于注射后6h腹腔注射荧光素酶底物,并通过活体动物体内光学成像系统检测基因表达的初始水平。将辐照组小鼠进行700c Gy非清髓性全身照射,并于水流动力学注射后12、24、48、72、96、120 h通过活体动物体内光学成像系统对两实验组小鼠体内目的基因的表达水平进行监测。结果通过尾静脉进行水流动力学注射的PGL3-luc质粒在注射后6h于小鼠肝脏大量表达,其平均表达水平为(3739±924.8)相对荧光单位(RLU)/(sec·mg)蛋白。在各个时间监测点辐照组与非辐照组相比实验鼠体内基因表达水平差异均有统计学意义(均P>0.05),其表达水平均于水流动力学注射72 h后显著降低。结论小鼠移植前的辐照预处理对水流动力学注射介导的基因在小鼠体内的表达并无影响,该种新型基因转移表达方法可以有效地应用于小鼠造血干细胞移植模型中。

Objective To explore the effects of irradiation on hydrodynamic injection-mediated luciferase reporter gene expression in mice and evaluate the gene transfer efficiency using HGT while applied to murine hematopoietic stem cell transplantation model.Methods The mice were divided into the irradiated group and the non-irradiated group.The PGL3-luc plasmid expressing luciferase reporter gene was injected into mice by a rapid large-volume injection.Six hours after the injection the luciferase substrate was intraperitoneal injected into mice and the initial level of gene expression was detected through in vivo imaging system.The irradiated group mice received nonmyeloablative doses（700c Gy） of total body irradiation （TBI）.The level of gene expression in two groups of mice was then monitored by in vivo imaging system in real time at the 12th,24th,48th,72nd,96th,and 120th h after the hydrodynamic injection. Results High expression of the transgene delivered via the tail vein was observed in the mouse liver at the sixth hour after injection and the average expression level of（3739±924.8） RLU/（sec·mg） protein was obtained.The level of transfer gene expression was similar between mice in the irradiated group and those in the non-irradiated group at various time（P〉0.05） but fell to a low level three days after gene transfer.Conclusion The irradiation after the hydrodynamic injection doesn＇ t affect the level of transfer gene expression in mice.This new type of gene transfer methods can be applied to mouse hematopoietic stem cell transplantation model effectively.