摘要
目的:比较腺病毒、腺相关病毒和脂质体-质粒三种载体在肌腱中的基因转染效率并观测腺相关病毒载体介导bFGF基因在肌腱愈合过程中表达。方法:用微量注射器将携带增强绿色荧光蛋白报告基因的三种载体注射入正常肌腱中,用荧光显微镜观测术后不同时间点肌腱内EGFP的表达。将AAV2-bFGF病毒颗粒注入损伤肌腱,使用免疫组化染色观测bFGF在肌腱内表达。结果:三种不同的载体转染肌腱3天后有EGFP表达、7天最明显、14天时下降;21天时少有表达。在同期的时间点,注射腺病毒、腺相关病毒组肌腱内EGFP表达明显强于质粒载体,而腺病毒、腺相关病毒两组之间无较大区别。免疫组化显示注射AAV2-bFGF后的第2、3和4周的肌腱表达很强的bFGF。结论:腺病毒、腺相关病毒载体的基因转染效率高于脂质体-质粒载体,AAV2携带的外源性bFGF基因在肌腱细胞内较强表达,提示我们将来在体内转基因到肌腱的研究中能够选择腺病毒和腺相关病毒载体作为运载基因的工具。
Objective： To investigate the transfection efficiency of adenoviral, adeno-associated viral and liposome-plasmid vectors in tendons, and explore bFGF expression in tendon mediated by AAV2 vectors. Methods： pCMV-EGFP, AAV2-EGFP and Ad5-EGFP were injected into digital flexor tendons of the chickens respectively, while EGFP expression were explored under the fluorescence microscope. AAV2-bFGF was injected into the injured digital flexor tendons, followed by immunohistochemical staining for detecting bFGF expression in tendons. Results： After 3 day＇s transfecting, EGFP expression was observed in tendons, with the strongest expression at the 7 days, regardless of different vectors. EGFP expression decreased at 14 days and disappeared at 21 days. The EGFP expression level in tendon injected with AAV2-EGFP and AdS-EGFP was higher than that of pCMV-EGFP injection. We did not find remarkable expression difference between AAV2-EGFP and Ad5-EGFP vectors. The AAV2-bFGF treatment remarkably improved bFGF expression in tendons at 2, 3 and 4 weeks after injection, which were proved by immunohistochemieal staining. Conclusions： Transfection efficiency of AAV2 and Ad5 vectors are higher than that of liposome-plasmid vectors. The tendons treated with AAV2-bFGF express bFGF protein strongly, which suggests that AAV2 and adenoviral vectors are promising gene delivery vectors for tendon gene therapy.
出处
《中国临床解剖学杂志》
CSCD
北大核心
2009年第2期195-198,共4页
Chinese Journal of Clinical Anatomy
基金
国家自然科学基金(30571882)
江苏省"135工程"重点学科科研课题基金(SK2002-21)
江苏省333工程基金
江苏省手外科临床医学中心建设基金
关键词
基因载体
绿色荧光蛋白
基因表达
腺相关病毒
肌腱组织
免疫组化
genetic carrier
green fluorescent protein
gene expression
adeno-associated viral vector
tendon
immunohistochemistry
