流体动力学法乙型肝炎病毒感染动物模型的建立

目的:建立一种简便、有效、稳定的乙型肝炎病毒(HBV)感染的动物模型,观察该模型动物体内不同时间点各种HBV标志物的表达情况．方法:以流体动力学法尾静脉注射BALB／c 小鼠pcDNA3．1-HBV,1 wk内检测各种HBV 标志物,时间分辨免疫荧光分析法(IFMA)检测血清中HBsAg,HBeAg,抗HBs,抗HBe,抗 HBc,荧光定量PCR法(FQ-PCR)检测血清HBV DNA,免疫组织化学法检测肝组织HBsAg和 HBcAg．结果:成功建立一种急性HBV感染动物模型, 第1天血清中HBsAg表达达高峰,后逐渐降低, HBeAg表达量少,分别在第4、5、7天检测到抗HBc,抗HBe,抗HBs,d1 HBV DNA滴度亦达高峰,后渐下降,免疫组织化学示HBsAg呈胞质内弥漫性分布,HBcAg亦主要为胞质型分布．结论:以流体动力学法建立的HBV模型是一种新型有效的HBV感染动物模型,能稳定较高水平表达大部分HBV标志物．

Development of a hepatitis B virus infection animal model by hydrodynamics

AIM: To develop a simple, convenient, efficient and stable animal model of hepatitis B virus (HBV) infection, and observe the dynamic changes of viral replication as well as expression in the model. METHODS: pcDNA3.1-HBV was delivered into BALB/c mice using the hydrodynamic tail vein injection method. In a week, the levels of HBsAg, HBeAg, anti-HBs, anti-HBe, and anti-HBc were determined with rime-resolved immunofluorometric assay (IFMA) kit, and the titers of HBV DNA were analyzed by fluorogenic quantitative polymerase chain reaction (FQ-PCR). In addition, viral specific proteins (HBsAg and HBcAg) in the liver were assayed by immunohistochemical staining. RESULTS: A mouse model of acute HBV infec- tion was developed successfully. HBsAg expression reached the peak on day 1, and dropped gradually. The level of HBeAg was low, and the level of anti-HBc,anti-HBe, and anti-HBs were first detected on day 4, 5 and 7, respectively. The titers of HBV DNA also peaked on day 1 and declined thereafter. HBsAg was positively scattered in the cytoplasm of hepatocytes, and HBcAg was mainly positively expressed in the cytoplasm of hepatocytes, too. CONCLUSION: This is a novel and effective animal model of HBV infection, in which HBV can replicate and express stably.