腺病毒载体转染人大隐静脉平滑肌细胞效率的研究  

Study of improvement of efficiency of gene transfer to smooth muscle cell in cultured human saphenous vein

作　　者：罗涛[1] 张强[1] 刁彦鹏[1] 段佩琰[1] 罗英伟[1] 段志泉[1] 

机构地区：[1]中国医科大学附属第一医院血管外科,沈阳110001

出　　处：《中华实验外科杂志》2003年第3期209-210,T001,共3页Chinese Journal of Experimental Surgery

摘　　要：目的　探讨提高静脉壁通透性对腺病毒为载体向人大隐静脉平滑肌细胞转外源基因效率的影响。方法　人体大隐静脉经机械拉伸、腔内加压灌注携 β 半乳糖酶基因的腺病毒溶液。器官培养 0、2d后 ,进行免疫组织化学染色和扫描电镜观察。研究机械因素对静脉组织形态的改变、鉴定转基因靶细胞成份、比较各组平滑肌细胞的转染率。 14d时 ,进行核增殖抗原免疫组织化学和弹力纤维染色 ,比较各组平滑肌细胞增殖指数和内膜增生程度 ,观察机械因素对培养静脉的远期影响。结果　机械牵拉和加压灌注后 ,静脉形态结构完整 ,内皮未受损伤 ,细胞间出现微小孔隙。中膜平滑肌细胞的转染率达到 ( 2 1.1± 3 .8) %。对照组静脉内皮细胞间无间隙 ,仅 ( 0 .8± 0 .2 ) %中膜平滑肌细胞表达 β 半乳糖酶。培养 14d后平滑肌细胞增生指数和内膜增生程度在各组间差别无显著性。结论　提高静脉通透性可提高腺病毒载体对中膜平滑肌细胞的转基因效率。Objective To investigate the effect of increased permeability of vein graft on efficiency of gene transfer to smooth muscle cells.Methods By longitudinal stretch and physiological pressure on human saphenous vein,adenovirus vector carrying LacZ gene was infused.Organ culture of veins was developed.0 or 2 days later,the samples were subjected to histochemical staining or observed under scanning electron microscopy.The cell type stained in the vein was qualified and indexes of efficiency of gene transfer were compared between groups.Miller's elastic van Gieson and PCNA staining were carried out to assess the thickness of neointima formation and medial smooth muscle cells proliferation,which indicated the influence of mechanical factor on saphenous veins.Results The porosity appeared among endothelial cells without denudation.The expression of mark gene in medial smooth muscle cells was up to (21.1±3.8)%.The porosity of control groups had no change and the index of gene transfer was (0.8±0.2)%.Mechanical factors did not affect the intimal hyperplasia and smooth muscle cell proliferation.Conclusion Augmentation of vein permeability could improve efficiency of gene transfer with adenovirus vector.

关 键 词：腺病毒载体 基因转染 人大隐静脉 平滑肌细胞效率 研究 免疫组织化学 自体静脉移植 

分 类 号：R654.3[医药卫生—外科学]