中文摘要：

目的：开发超声裸基因药物传递与治疗新方法，研究适合超声基因传递的安全计量与参数，进行安全性评价，建立超声裸基因传递系统。方法：建立一套经水听器与声功率计标定的低频超声基因传递实验系统（35．1K），用不同的计量对兔、鸡、S180肿瘤细胞悬液进行暴露，提取最佳基因传递参数。用绿色荧光蛋白作为传递基因进行传递实验。用荧光法、分光光度法、扫描电镜法、激光共聚焦扫描法和流式细胞术评估实验结果。结果：最佳传递参数下低频超声有效地将GFP基因传入了S180细胞，未见明显的细胞损伤与细胞毒性，转染率为35.83％（±2．53，n=6），并经活体证明细胞功能正常。90％细胞存活点的能量积累E是最佳传递参数（E=3．56±0．06），80％细胞存活点的E值是损伤闽值（E=59．67±3．54）。GFP的表达明显比病毒载体法强（P〈0．001）。结论：超声可安全地用于基因传递和治疗，其基因表达和摄入量取决于90％细胞存活点的能量积累E。它可用作控制因子与其它参数结合控制传递效应。从结果证明平稳空化产生基因传递效应，瞬间空化造成细胞损伤。

英文摘要：

Object： The purpose of this work is to study a novel method of physical ultrasound （US） gene delivery and therapy investigate safe energy and parameters and set up a experimental system of ultrasonic naked gene delivery system. Methods： The ultrasound experimental system was built and the power was measured by an acoustic power meter calibrated by hydrophone. The red blood cells of rabbit and chicken and mice S180 cells were exposed to US with different exposure time and intensity. Fluorescent microscope, spectrophotometer, scanning electron microscope, laser scanning confocal microscope and flow cytometry were used to investigate the optimal parameters. The plasmid of green fluorescent protein served as reporter gene. Results： US （35.1 kHz） effectively delivered GFP gene into S180 cells at optimal parameters without evident damage and cytotoxity in vitro. The transfection rate was about 35.83 % （±2.53, n = 6） in viable cells. The function of exposed cells proved normal in vivo. The energy accumulation E of US delivery at 90% cell viability was optimal parameters （E = 3.56±0.06）, and 80% of cell viability was damage threshold （E = 59.67±3.54）. The intensity of GFP expression showed a higher fluorescent peak over AVV-GFP group and control group （p 〈 0.001）. Conclusion： With optimal parameters, the quantitative US can safely deliver naked gene into cells without the damage of cell function. Both optimal uptake and expression of gene depend on the energy accumulation E. As a control factor, E can be employed to control bioeffect combining with other parameters. The stable caviation yields to optimal parameters of gene delivery and the transient caviation may cause cell damage. The results were given as basic data to develop novel clinical gene therapeutic system.