关键词: GFP表达质粒；流体力学注射；门静脉；基因疗法

Abstract:

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 GFPplasmids 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.

Key words: GFP plasmid; Hydrodynamic injection; Portal vein; Gene therapy