心脏移植中经冠脉系统转基因的实验研究

Experimental study on ex vivo gene transfer intracoronarily to donor heart

目的 研究将外源基因转入移植心脏 ,进行基因治疗的可行性。方法 建立小鼠异位心脏移植模型 ,建立经冠脉循环系统灌注技术。将 12只BAIB/c近交系小鼠随机分为转基因组和空白对照组 ,每组各 6只 (供、受者各 3只 )。转基因组经冠脉循环系统将携带报告基因 半乳糖苷酶基因 (Lac Z)的质粒载体 (PSV β gal) /脂质体 (DOSPER)混合物灌注入供心 ,而后进行异位心脏移植 ;对照组则灌注生理盐水。术后 3d获取供心 ,作冰冻切片 ,经X gal组织化学染色检测Lac Z基因的转染和表达情况。结果 转基因组的 3例供心都检测到Lac Z基因的表达 ;其中有 2例Lac Z基因表达位于心室壁中层的心肌内 ,1例其表达位于心内膜下的心肌内。而在对照组则没有检测到Lac Z基因。结论 以冠脉循环系统灌注的方式将质粒载体 /脂质体的混合物灌注入供心 。

ObjectiveTo study the feasibility of ex vivo gene transfer to donor heart by delivering i ntracoronarily a reporter gene (Lac-Z gene) to donor heart at the time of trans plantation.MethodsThe model of heterotopic heart transplantation in murine was established and the method of perfusing intracoronarily was performed. Twelve male BALB/c mice were divided into 2 groups randomly: gene-transfer group (group 1) and control grou p (group 2). In group 1, plasmid vector (PSV-β-gal containing Lac-Z gene)/li posome (DOSPER) was perfused intracoronarily into each donor heart at the time o f transplantation. In group 2, normal saline was perfused. Donor hearts were har vested 3 days after transplantation. Freezing sections were made for detection o f the transfer and expression of Lac-Z gene by histochemical staining (X-gal). ResultsThe expression of Lac-Z gene was detected in the donor hearts of group 1. In tw o donor hearts, the expression of Lac-Z gene was detectable in the myocardial c ells in the mid-layer of ventriculus; In one donor heart the expression was fou nd in the myocardial cells under epicardium. But no expression of Lac-Z gene wa s detected in donor hearts of group 2.ConclusionEx vivo gene transfer intracoronarily to donor heart in the form of plasmid vector /liposome complex at the time of transplant ation is feasible.