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 investigate the optimal conditions of ultrasound-mediated microbubble-mediated gene transfer into rat Achilles tendon and granulation tissue. Methods The model of Achilles tendon injury was established in rats. The mixed solution of microbubbles and enhanced green fluorescent protein (EGFP) plasmids was injected locally into the injured Achilles tendon of rats. Different output powers, duty cycles and ultrasonic irradiation time Ultrasound was used to irradiate the Achilles tendon on both sides. The transfection efficiency was judged according to the results of fluorescence staining and immunohistochemical staining of EGFP. According to the highest gene transfection efficiency, while the smallest normal tissue necrosis screened the most appropriate irradiation conditions. The rats were divided into 4 groups: 1 plasmid + microbubble + ultrasound irradiation group, 2 plasmid + microbubble group, 3 plasmid + ultrasound irradiation group, 4 pure plasmid group. According to the conditions selected in the previous steps, the Achilles tendon was irradiated and the expression of EGFP in the Achilles tendon and granulation tissue and normal tissue damage were observed. Results The expression of EGFP in the Achilles tendon and granulation tissue was obvious under the conditions of ultrasonic power of 2 W / cm2, duty cycle of 20% and ultrasonic irradiation for 10 min. There was no obvious normal tissue injury. The expression of EGFP in the ultrasound irradiation + microbubble + plasmid group was higher than that in the other three groups (P <0.05), and there was no significant difference among the other groups. Conclusions Ultrasound combined with microbubbles can significantly enhance the gene transfection efficiency of injured tendon and granulation tissue without damaging the normal tissue under suitable ultrasonic irradiation conditions, which provides the experimental basis for the feasibility of gene therapy for tendon injury.