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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 investigate the expression of the target gene in mouse liver and its transgene efficiency after injection of GFP plasmid by three different routes of fluid mechanics, portal vein and routine intraperitoneal injection. Methods Naked plasmid or liposome-encapsulated plasmid DNA was injected into allogeneic mice by hydrodynamic injection, portal vein and routine intraperitoneal injection respectively. After 48 hours, blood and liver tissues were taken out and the three transfection routes were observed by fluorescence microscopy On the expression of plasmid DNA in mouse liver. Results A large amount of green fluorescent protein (GFP) was detected in both hydrodynamic and portal venous injection groups. There was no significant difference in fluorescence expression between the two groups (P> 0.05). Only a small amount of green fluorescence was observed in the liver of mice injected intraperitoneally The expression of green fluorescence in the three groups of liposome / plasmid DNA complex was significantly higher than that of the naked plasmid group (P <0.05). Conclusion Both hydrodynamic injection and conventional portal vein injection of liposome / plasmid DNA complex, the target genes are highly expressed in the liver of mice, no significant difference between the two ways, hydrodynamic injection can be widely used in liver-targeted live genes Transfection.