树鼩水动力转染方法的建立及在HBV模型探索中的应用

Tree shrew model of acute hepatitis B virus infection generated by hydrodynamic injection

目的建立一种水动力注射将外源基因导入树鼩肝脏的方法,并用此方法建立HBV树鼩模型。方法用萤火虫荧光素酶(firefly luciferase,Fluc)和β-半乳糖苷酶(β-galactosidase,β-Gal)作为报告基因,通过树鼩大隐静脉将报告基因用水动力注射方法导入树鼩肝脏,活体成像和β-Gal染色观察水动力转染效果。将pHBV1.2质粒用水动力注射方法转入树鼩肝脏,检测树鼩血清中ALT、实时荧光定量检测血清中HBV DNA,ELISA检测血清中HBsAg、HBsAb。结果通过大隐静脉水动力注射,报告基因能够在肝脏特异性表达,转染HBV1.2质粒后树鼩血清中能够检测到相关阳性指标。结论大隐静脉水动力注射法能够作为树鼩肝脏外源基因导入的一种方法,并用此方法初步探索了树鼩的HBV模型。

ObjectiveTo establish a method for hydrodynamic-based transfection to tree shrews and generate a tree shrew model of hepatitis B virus infection.MethodsThe firefly luciferase（Fluc） and a β-galactosidase（β-Gal） as reporter genes were delivered to hepatocytes of tree shrews by hydrodynamic-based injection via the great saphenous vein.Bioluminescence imaging and β-Gal staining were used to verify the gene expression.The tree shrew was hydrodynamically injected with pHBV1.2 plasmid.At various times,the sera were collected in which ALT,HBsAg,HBsAb and HBV DNA were examined by ELISA or real-time fluorescence quantitative PCR respectively.ResultsThe reporter genes were detected only in the liver of tree shrews by bioluminescence imaging and β-Gal staining.Markers of HBV were detected in the sera of tree shrews injected hydrodynamically with pHBV1.2.ConclusionHydrodynamic-based injection by the great saphenous vein can be used to deliver the gene to hepatocytes of tree shrews.A tree shrew model of acute HBV infection is generated by the hydrodynamic injection.