超声联合微泡促进大鼠损伤跟腱及肉芽组织局部基因转染的实验研究  

Enhanced Local Gene Transfection in Rats with Injured Achilles Tendons and Granulation Tissues using Ultrasound and Microbubble

作　　者：唐远姣[1] 向茜[1] 冷钱英 张凌燕[1] 邱逦[1] 

机构地区：[1]四川大学华西医院超声科,成都610041

出　　处：《四川大学学报（医学版）》2014年第6期979-983,1009,共6页Journal of Sichuan University(Medical Sciences)

基　　金：国家自然科学基金(No.81271585)资助

摘　　要：目的探讨超声联合微泡促进基因局部转染大鼠损伤跟腱及肉芽组织的最适超声转染条件。方法建立双侧大鼠跟腱损伤模型,在大鼠双侧损伤跟腱内局部注射微泡与增强型绿色荧光蛋白(EGFP)质粒混合溶液,应用不同输出功率、占空比及超声辐照时间的超声辐照双侧损伤处跟腱,根据荧光染色及免疫组化染色下EGFP表达结果判断转染效果。根据基因转染效果最高,同时正常组织损伤坏死最小筛选出最合适的照射条件。将大鼠分为4组:1质粒+微泡+超声辐照组,2质粒+微泡组,3质粒+超声辐照组,4单纯质粒组。根据前几步所选取的条件辐照损伤跟腱,观察跟腱及肉芽组织内的EGFP表达情况及正常组织损伤情况。结果在超声输出功率2 W/cm2、占空比20%、超声辐照10min条件下,跟腱及肉芽组织内EGFP表达明显,且无明显正常组织损伤。超声辐照+微泡+质粒组跟腱及肉芽组织内EGFP表达高于其余3组(P<0.05),其余各组间差异无统计学意义。结论在适合的超声辐照条件下,超声联合微泡能明显增强损伤肌腱及肉芽组织的基因转染效果,且不损伤正常组织,这为肌腱损伤的基因治疗的可行性提供了实验基础。Objective To identify best local ultrasound transfection conditions in rats with injured achilles tendons and granulation tissues.Methods Rat models with bilateral injured achilles tendons were established.A mixture of microbubbles and enhanced green fluorescent protein（EGFP）plasmids were injected into the achilles tendons.The injured tendons were subject to ultrasound irradiation under different output intensity,duty cycle and irradiation time.The rats were divided into four groups：①plasmid＋microbubbles＋ultrasound（PL＋MB＋US）;②plasmid ＋ microbubble（PL＋MB）;③plasmid ＋ ultrasound（PL＋US）;④ plasmid only（PL）.We identified the best ultrasound conditions based on the highest transfection efficiency in the achilles tendons and granulation tissues and the least normal tissue damage.Results Under the condition of 2 W/cm^2 power output and 20% duty cycle of ultrasound irradiation for 10 min,EGFP expression was demonstrated in the achilles tendons and granulation tissues without showing significant damage to normal tissues.Higher levels of EGFP expression were found in the group of PL＋ MB＋US than the other groups（no significant differences in EGFP expression were found among the other three groups）.Conclusion Under proper conditions,ultrasound and microbubble can enhance in vivo gene transfection efficiency in rats with injured achilles tendons and granulation tissues without causing obvious normal tissue damage,which could provide a sound basis for experimental study of gene therapy for injured tendons.

关 键 词：超声微泡 基因转染 损伤跟腱 肉芽组织 

分 类 号：R445.1[医药卫生—影像医学与核医学] R686.1[医药卫生—诊断学]