腺病毒对内皮及成纤维细胞和血管环转染及基因表达的研究  

Adenovirus - mediated Gene Transfer and Expression in Endothelial Fibroblasts Cells and Vascular Graft

作　　者：穆军升[1] 郑家豪[1] 朱洪生[1] 山朝晖[1] 许德华[2] 刘新垣[2] 

机构地区：[1]上海第二医科大学仁济医院心胸外科,上海200001 [2]中国科学院上海分院生化细胞研究所

出　　处：《上海第二医科大学学报》2002年第3期221-224,共4页Acta Universitatis Medicinalis Secondae Shanghai

基　　金：卫生部科学研究基金资助(98-2-295)

摘　　要：目的 比较腺病毒对不同血管内皮及成纤维细胞的转染和对人冠状动脉搭桥动脉静脉血管的转导,为携带目的基因的腺病毒治疗血管狭窄提供更好的转染途径。方法 腺病毒Ad5CMVLacZ体外转染血管内皮细胞（HUVEC）和成纤维细胞（PA317）并间接转导入胸廓内动脉和大隐静脉血管环,运用β-半乳糖苷酶活性X-gal组织化学染色,观察转染情况,并进行动态分析。结果 腺病毒载体能使外源性基因有效地转入HUVEC、PA317及两种血管环并获得蛋白表达;两种血管环在转染10d后仍有β-半乳糖苷酶蛋白的表达,且大隐静脉有新内膜形成和中膜的增厚。结论 从细胞水平上证明腺病毒载体能将治疗性目的基因导入各种血管细胞及动静脉血管并获得表达,提示血管内膜和外膜都是腺病毒有效转染的途径;临床上无论动脉或静脉的血管狭窄都可能通过用腺病毒作为载体进行基因治疗和预防。Adenovirus - mediated gene transfer in the artery and vein is a promising pathway in the study of vascular biology and the development of vascular gene therapy. The efficiency of gene transfer by means of deliverling adenovirus vectors via the adventitia was studied in arterial and venous graft. Methods Replication - defective adenovirus vectors encoding a β- galactosidase(β -gal) gene(Ad5CMVLacZ)or encoding endothelial nitric oxide synthase gene (Ad5CMVeNOS) were used in vitro fibroblasts cell and endothelial cells, and in ex vivo tissues of the human internal mammary artery and saphenous vein of 6 human beings who underwent graft operation. Cells or samples were used for β- galactosidase histochemical staining. Dynamic analyses were done. Results Histochemical staining demonstrated extensive blue coloration areas in the intima and adventitia of each graft. Expression of X- gal still existed after transduction for 10 days. Fibroblasts and endothelial cells were stained too. Conclusion Adenovirus vector could make LacZ gene effectively transfer into the graft to attain a high level of gene expression.

关 键 词：腺病毒 内皮 成纤维细胞 血管环转染 基因表达 研究 

分 类 号：R654.2[医药卫生—外科学]