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Establishment and primary application of a mouse model with hepatitis B virus replication

机译:乙型肝炎病毒复制小鼠模型的建立和初步应用

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摘要

AIM: To establish a rapid and convenient animal model with hepatitis B virus (HBV) replication.METHODS: A naked DNA solution of HBV-replication-competent plasmid was transferred to BALB/C mice via the tail vein, using a hydrodynamic in vivo transfection procedure. After injection, these mice were sacrificed on d 1, 3, 4, 5, 7 and 10. HBV DNA replication intermediates in the liver were analyzed by Southern blot hybridization. The expression of hepatitis B core antigen (HBcAg) and hepatitis B surface antigen (HBsAg) in the liver was checked by immunohistochemistry. Serum HBsAg and hepatitis B e antigen (HBeAg) was detected by enzyme-linked immunosorbent assay (ELISA). Inhibition of HBV replication was compared in HBV replication model mice treated intraperitoneally with polyinosinic-polytidylin acid (polyIC) or phosphate-buffered saline (PBS).RESULTS: After hydrodynamic in vivo transfection, HBV DNA replication intermediates in the mouse liver were detectable on d 1 and abundant on d 3 and 4, the levels were slightly decreased and remained relatively stable between d 5 and 7, and were almost undetectable on d 10. The expression patterns of HBcAg and HBsAg were similar to that of HBV replication intermediate DNA, except that they reached a peak on d 1 after injection. No obvious differences in HBV DNA replication intermediates were observed in the left, right and middle lobes of the liver. After treatment with polyIC, the level of HBV intermediate DNA in the liver was lower than that in the control mice injected with PBS.CONCLUSION: A rapid and convenient mouse model with a high level of HBV replication was developed and used to investigate the inhibitory effect of polyIC on HBV replication, which provides a useful tool for future functional studies of the HBV genome.
机译:目的:建立具有乙型肝炎病毒(HBV)复制功能的快速便捷的动物模型。方法:通过水动力体内转染,将含HBV复制功能的质粒的裸DNA溶液通过尾静脉转移至BALB / C小鼠。程序。注射后,在第1、3、4、5、7和10天处死这些小鼠。通过Southern印迹杂交分析肝脏中的HBV DNA复制中间体。通过免疫组织化学检查肝脏中乙型肝炎核心抗原(HBcAg)和乙型肝炎表面抗原(HBsAg)的表达。通过酶联免疫吸附试验(ELISA)检测血清HBsAg和乙型肝炎e抗原(HBeAg)。结果比较:经水动力体内转染后,小鼠肝脏中可检测到HBV DNA复制中间体。 1和d 3和4时丰富,在d 5和7时水平略有下降并保持相对稳定,而在d 10时几乎未检测到。HBcAg和HBsAg的表达模式与HBV复制中间体DNA相似,除了他们在注射后第1天达到峰值。在肝脏的左,右和中叶没有观察到HBV DNA复制中间体的明显差异。经polyIC处理后,肝脏中的HBV中间DNA水平低于注射PBS的对照组。结论:建立了快速便捷的HBV复制水平高的小鼠模型,并研究了其抑制作用IC对HBV复制的研究,为HBV基因组的未来功能研究提供了有用的工具。

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