经心包腔途径转染提高大鼠体内心肌细胞转染的效率

Improvement of transfection efficiency with plasmid transfected to the myocardium of rats intrapericardially in vivo

背景:如何获得安全、有效、广泛心肌组织的转染一直是国内外学者研究的热点。目的:探讨能够改善动物水平心肌组织非病毒载体的转染效率、增强基因导入靶向性的方法。方法:以β半乳糖苷酶质粒PLacZ作为报告基因,将Wistar大鼠随机分为5组:①空白组。②心包腔内组:心包腔注射质粒+微泡+酶混悬液,超声导入。③心包腔内阴性对照组:以生理盐水代替质粒干预。④舌下静脉组:舌下静脉注射质粒+微泡混悬液,超声导入。⑤舌下静脉阴性对照组:以生理盐水代替质粒干预。注射6d后处死,进行心、肺、肝、肾组织的X-gal染色。结果与结论:转染6d后,仅有心包腔内组大鼠的部分心肌细胞在心尖、心室及心房水平可见明显的蓝染,其他各组大鼠心肌X-gal染色为阴性;各组大鼠肺、肝、肾组织X-gal染色均为阴性。提示采用心包腔内途径转染、再辅以超声微泡导入以及酶类的使用,可明显改善质粒对在体心肌细胞的转染效率,且不伴有心外组织目的基因的表达,具有较好的靶向性。

BACKGROUND：It has always been the highlights to find how to accomplish safe,effective and extensive transfection of the myocardium.OBJECTIVE：To explore a new feasible method for improving transfection efficiency of non-viral plasmids to the myocardium and for obtaining satisfactory myocardial targeting.METHODS：We chose PLacZ as a report gene.Wistar rats were randomly divided into five groups：control group,intrapericardial group,intrapericardial negative group,sublingual vein group,sublingual vein negative group.For all the subsets of experiments,hearts were harvested at 6 days after injection.Tissues of heart,lung,liver and kidney were stained with X-gal,and PlacZ gene expression of the heart and other non-targeted organs was observed to decide the gene transfection status and myocardial targeting.RESULTS AND CONCLUSION：After 6 days of transfection,only rats in the intrapericardial group were stained blue in parts of the myocardium on the atrial,ventricular and apex level.Other groups were negative for X-gal staining in the myocardium,lung,liver and kidney.The intrapericardial injection of microbubbles and some enzymes,as well as plasmids,with the aid of ultrasound,can improve transfection efficiency of plasmids significantly and the heart targeting is satisfactory.