CAJ | 학술논문

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.
목적 탐토초성복조미포조영제성낙유(SonoVue)능부증강rAAV2-EGFP전염시망막적효솔.방법 62지Wistar대서진행파리체강(10지)급시망막하(52지)주사.실험분조:병독+생리염수、병독+미포、병독+생리염수+초성복조화병독+미포+초로복조.제4、7、35、49、120 d분별용형광체시경관찰,이용연건진행정량분석,병진행조직절편광경검사.결과 파리체강주사조지속관찰2개월미견형광;시망막하주사조적초성복조미포조재제4、7、35 d교기타조전염효솔고,초성복조미포조영제대안저조직무명현손상.결론 초성복조SonoVue가재체내촉진rAAV2-EGFP전염대서시망막.
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.