rAAV8-1.3HBV构建小鼠病毒性肝损伤模型研究

Construction of a Mouse Model of Viral Liver Injury by rAAV8-1.3HBV

目的:用高嗜肝性的重组8型腺相关病毒载体携带C基因型(adr亚型) 1.3拷贝乙型肝炎病毒,建立HBV持续复制的小鼠病毒性肝损伤模型。方法:将rAAV8-1.3HBV经尾静脉注射到78只C57BL/6小鼠体内,通过定量检测血清中HBV DNA、HBeAg、HBsAg含量,同时检测血清学AST、ALT活力值以及病毒注射后8周、12周、16周、20周、24周时取实验组动物肝脏做HE染色分析,最终建立HBV持续复制的小鼠病毒性肝损伤模型。结果:C57BL/6小鼠注射rAAV8-1.3HBV后,8周至24周小鼠血清中可检测出稳定表达的HBV,且20周和24周HE染色病理结果显示肝组织内可见肝小叶内点状、灶状以淋巴、单核细胞为主的炎细胞浸润,肝小叶内的炎性浸润灶数量较多,分布范围较广,与对照组相比,病变程度及累计范围明显加重。结论:利用rAAV8体内转导C57BL/6小鼠,成功地建立了HBV病毒性肝损伤小鼠模型,此模型在病毒注射20周后逐渐出现病毒性肝损伤特征,可用于病毒性肝损伤的研究。

Purpose:To infect mice with liver-tropic recombinant adeno-associated virus type 8(rAAV8)vector carrying C genotype(adr subtype)1.3 copies of Hepatitis B virus(HBV)to create a mouse model for HBV-induced liver injury.Methods:First,rAAV8-1.3HBV was injected into the tail vein of 78 C57BL/6 mice.Quantitative detections of HBV DNA,HBeAg and HBsAg in serum showed that a mouse model of HBV continuous replication was established.Serological examinations for AST,ALT activity values and liver tissue staining of the satellite animals were performed at 8 weeks,12 weeks,16 weeks,20 weeks,and 24 weeks following virus administration.Finally,a mouse model of viral liver injury with sustained replication of HBV was established.Results:After injection of rAAV8-1.3HBV in C57BL/6 mice,stable expression of HBV was detected in the serum of mice during the period from 8 weeks to 24 weeks,and the pathological results revealed by HE staining at 20 and 24 weeks showed visible lesions,foci,and lymphoid cells in hepatic lobules in the liver tissue accompanied by inflammation and monocyte invasion.There was a relatively high count with a wide distribution range of invasive cells in the hepatic lobules.Compared with the control group,the degree and total range of lesions were significantly elevated.Conclusion:The mouse model of HBV viral liver injury was successfully established by in vivo administration of rAAV8 into C57BL/6 mice.The created model showed the characteristics of the gradual development of liver injury suggestive of hepatic infection during the 20 weeks following virus administration.Therefore,this model may be used to study virus infection induced liver injury.