不同转染方式对GFP表达质粒在小鼠肝脏的表达影响

Comparison of different non-viral transfection methods for GFP plasmid expression in the liver of mice

目的研究经流体力学注射、门静脉和腹腔常规注射3种不同途径注射GFP表达质粒后,目的基因在小鼠肝脏的表达情况及其转基因效率。方法将裸质粒或脂质体包裹的质粒DNA分别应用流体力学注射、门静脉及腹腔常规注射法注入同种异体小鼠体内,48h后分别取血和肝组织,通过荧光显微镜观察3种转染途径对质粒DNA在小鼠肝脏的表达影响。结果流体力学注射组及门静脉常规注射组均可见大量绿色荧光蛋白表达,两组的荧光表达量差异无统计学意义(P>0.05),腹腔注射组小鼠的肝脏仅见少量的绿色荧光表达,但3组内脂质体/质粒DNA复合物组绿色荧光表达量均明显高于裸质粒组(P<0.05)。结论应用流体力学注射及门静脉常规注射脂质体/质粒DNA复合物途径,目的基因均在小鼠肝脏高效表达,两种途径无明显差异,流体力学注射可广泛用于肝靶向性的活体基因转染。

Objective To compare the transfection efficiency and expression intensity of a green fluorescent protein （GFP） reporter gene in the liver of mice by using three non-viral transfection methods,i.e.,hydrodynamic injection,portal vein injection and peritoneal injection. Methods The naked GFP plasmids or liposome encapsulated plasmids were injected via the tail vein or portal vein or abdominal cavity of different mice with homogeneity. The blood and the liver were harvested 48 h after injection,and the contents of alanine aminotransferase （ALT） and total bilirubin （TB） were detected in serum. Also,expression intensity of GFP was evaluated by fluorescence microscopy. Results Of the three transfection methods employed,hydrodynamic gene delivery and portal vein injection conferred a stronger expression of GFP with a similar transfection efficiency （P〉0.05）. However,after encapsulation by liposomes,the expression level of GFP was significantly higher than that of naked plasmid DNA. Conclusion High level gene expression in mouse liver can be achieved by hydrodynamic injection or portal vein injection of liposome encapsulated plasmid DNA,and hepatic delivery of the foreign gene can be accomplished by hydrodynamics-based injection.