摘要
目的 探讨超声辐照微泡造影剂声诺维（SonoVue）能否增强rAAV2-EGFP转染视网膜的效率.方法 62只Wistar大鼠进行玻璃体腔（10只）及视网膜下（52只）注射.实验分组：病毒＋生理盐水、病毒＋微泡、病毒＋生理盐水＋超声辐照和病毒＋微泡＋超卢辐照.第4、7、35、49、120 d分别用荧光体视镜观察,利用软件进行定量分析,并进行组织切片光镜检查.结果 玻璃体腔注射组持续观察2个月未见荧光;视网膜下注射组的超声辐照微泡组在第4、7、35 d较其他组转染效率高,超声辐照微泡造影剂对眼底组织无明显损伤.结论 超声辐照SonoVue可在体内促进rAAV2-EGFP转染大鼠视网膜.
Objective To investigate the practical efficacy and safety of ultrasound with microbubbles mediated rAAV2-EGFP to retina of rat after intravitreal and subretinal injection. Methods Gene transfer was examined by rAAV2-EGFP intravitreal and subretinal injection into the Wistar rats with or without microbubbles. The eyes were exposed to US （1 MHz,2 W/cm2, duration 5 minutes,duty cycle 50%,pulse recurrent frequency 100 Hz）. The onset of EGFP gene expression, lightness of fluorescence, area of fluorescence and its distribution in the fundus in vivo via fluorescence stereosocope were investigated on the 4th,7th, 35th,49th and 120th day respectively. The value of gene transfer was quantified through the EGFP fluorescence quantitative methods by Axiovision 3. 1 software. HE staining was used to observe tissue damage. Results There was no fluorescence observed by fluorescence stereosocope after intravitreal injection after two-month study. After subretinal injection, ultrasound-targeted microbubbles destruction （UTMD） strongly increased gene transfer efficiency. UTMD used in the experiment did no harm to the rat retina structure. Conclusions UTMD could not enhance rAAV2-EGFP transfecion efficiency to rat retina after intravitreal injection but the transduction could be enhanced significantly after subretinal injection.
出处
《中华超声影像学杂志》
CSCD
北大核心
2010年第5期440-442,共3页
Chinese Journal of Ultrasonography
基金
国家自然科学基金（30772369）
关键词
超声疗法
视网膜
转染
腺病毒科
Ultrasonic therapy
Retina
Transfection
Adenoviridae
