超声微泡造影剂介导EGFP质粒转染大鼠视网膜的实验研究

Experimental study on transferring EGFP gene into the retina of rat mediated by microbubbles

目的探讨超声破坏微泡介导EGFP质粒转染大鼠视网膜的效率及可行性,为实现外源基因高效、定向的转移目的奠定基础。方法将30只Long-evans大鼠分为6组,第1组仅以0.5W/cm2的超声波辐照大鼠眼球,第2组于尾静脉输入适当剂量的微泡造影剂,并立即以相同能量的超声波辐照大鼠眼球,第3组于尾静脉输入质粒,第4组于尾静脉输入质粒,并以超声辐照大鼠眼球,第5组于尾静脉输入质粒与微泡,第6组尾静脉输入质粒、微泡,并用超声辐照眼球。转染2周后,在激光共聚焦显微镜下观察EGFP表达情况。结果超声微泡介导的EGFP质粒对大鼠视网膜的转染效率,明显高于其他实验组。一定能量和时间的超声波辐照,及适当浓度的微泡,对大鼠视网膜脉络膜无明显损伤。结论利用低频率和一定能量的超声击碎携带EGFP质粒的超声微泡造影剂,能够有效地提高EGFP质粒在大鼠视网膜的转染效率。

Objective To investigate whether ultrasound-mediated microbubble destruction could effectively deliver plasmid to the retina of rat, and to afford a base for effective and directional gene delivery. Methods Thirty Long-evans rats were randomly divided into six groups. Microbubbles were infused into the tail vein of rats （group one and group two） with or without destructing microbubbles by ultrasound immediately. The naked plasmid DNA of EGFP without microbubbles was infused into the tail vein of rats （group three and group four） with or without destructing microbubbles by ultrasound immediately. Microbubbles attached with the naked plasmid DNA of green fluorescent protein （EGFP） were infused into the tail vein of rats （group five and group six） with or without destructing microbubbles by ultrasound immediately. After 2 weeks, the EGFP expression in the rats＇ retina and choroids were detected by confocal laser scanning microscopy. Results The infection efficiency of the sixth group were higher than other groups. Ultrasound the experiment had used did no harm to the mice＇s retina and never did the microbubbles. Conclusion The EGFP expression in rats＇ retina were increased with the administration of ultrasound-mediated microbubbles destruction.