超声介导击破微泡增强型绿色荧光蛋白正常大鼠关节滑膜组织

摘要

目的 探讨超声击破微泡介导增强型绿色荧光蛋白(EGFP)转染大鼠关节滑膜组织的可行性.方法 将20只正常清洁级Wister大鼠分为4组,每组5只(10个膝关节).单纯质粒组:10 μg EGFP注射入大鼠关节腔内；超声+质粒组:EGFP注射入大鼠关节腔内,超声持续照射10 min;微泡+质粒组:300μl SonoVue与10 μg EGFP混合注射入大鼠关节腔内；超声+微泡+质粒组:将300μl SonoVue与10 μg EGFP混合后注射入大鼠关节腔内,超声持续照射10 min.3天后取出大鼠膝关节滑膜组织,直接贴在载玻片上,在荧光显微镜480 nm波长激发状态下观察EGFP转染效果.结果 单纯质粒组、超声+质粒组和微泡+质粒组的大鼠膝关节滑膜组织EGFP的荧光强度稍有增强；超声+微泡+质粒组EGFP的荧光强度明显增强.结论 超声介导微泡可以实现EGFP质粒对正常大鼠关节滑膜组织的转染.%Objective To explore the feasibility of enhanced green fluorescent protein (EGFP) transfection into normal rat's knee joint synovial membrane by ultrasound-mediated microbubble destruction. Methods Twenty rats were divided into 4 groups (each including 5 rats/10 knees). I. E. EGFP group (injecting 10 μg EGFP into rats joints), ultrasound-)-EGFP group (irradiating with ultrasound for 10 min after 10 μg EGFP into rats joints). Microbubbles+EGFP group (injecting 300 μl SonoVue and 10 μg EGFP into rats joints) and ultrasound+microbubbles+EGFP group (irradiating with ultrasound for 10 min after 300 μl SonoVue and 10 μg EGFP into rats joints). Rats were sacrificed after 3 days and knee joints synovial membrane tissues were observed for EGFP protein expression on fluorescence microscopy. Results Light EGFP expression was seen in synovial membrane tissues of EGFP group, ultrasound + EGFP group and microbubbles+EGFP group, while strong EGFP expression was seen in synovial membrane tissues of ultrasound+microbubbles+EGFP group. Conclusion It is possible to deliver EGFP into rat's joint synovial membrane tissues by ultrasound-mediated microbubble destruction.