腺病毒载体携外源基因转染人培养静脉的实验研究

Adenovirus-Mediated Transfer of Xeno Gene Into Human Saphenous Vein Cultured in Vitro

目的 :探讨经腺病毒载体介导的标记基因转染人培养大隐静脉后 ,标记基因表达的效率、转染靶细胞表达时相。方法 :从临床手术患者取得大隐静脉后 ,浸于含AdCMV/LacZ或AdCMV的病毒液中 (5× 10 9pfu) ,孵育 1h ,将静脉剪成静脉片 ,分别培养 2 ,7,14d。对转基因静脉片进行组织学检查。结果 :腺病毒载体可有效地将标记基因 (LacZ基因 )转入培养大隐静脉。转染后 2d内皮细胞和外膜细胞即有表达 ,β 半乳糖苷酶组织化学染色可见胞核蓝染阳性细胞 ,7d时表达最高 ,持续表达到第 14d。对照大隐静脉无β 半乳糖苷酶表达。 结论 :腺病毒载体能高效转染人大隐静脉 ,其靶细胞以内皮细胞为主 ,外源性基因的表达至少维持

﨩bjective: [WT5'BZ]This study was to investigate the efficiency, cellular targets , time curve of expression of mark gene after ade novirus mediated transfer of gene into human saphenous veins in vitro. [WT5'HZ ]Methods: [WT5'BZ]The veins were obtained from patients and soaked in buffer l iquid containing AdCMV or AdCMV/LacZ (5×10 9 pfu)for 60 minutes. On the 2nd , 7th, 14th day after organ culture, histochemistry examination was performed. [ WT5'HZ]Results: [WT5'BZ]Adenovirus vector could transfer the mark gene (LacZ g ene) into cultured veins efficiently. After 2 days the cultured vein expressed g alactonidase in intima and adventitia. Peak expression of galactonidase occurred at the 7th day and remained elevated. No galactonidase expressed in controls. [ WT5'HZ]Conclusion: [WT5'BZ]Adenovirus vector can transfer gene into human vein s efficiently. The main target cells are endothelium cells. Gene expression sust ained for 2 weeks at least. [WT5'HZ]