乙肝病毒YMDD突变小鼠模型的建立及复制能力研究

Establishment of mouse model of hepatitis B virus YMDD mutation and study on replication capacity

目的:建立乙肝病毒YMDD突变小鼠模型,并对其体内HBV-DNA复制能力进行研究。方法:建立乙肝病毒多聚酶区(P区)rtM204I位点突变质粒,BALB/c小鼠分为两组,经尾静脉分别注射野生型和突变型HBV质粒,然后给予生理盐水(空白对照)和拉米夫定灌胃处理连续3 d。检测两组小鼠肝组织内HBV-DNA复制能力以及拉米夫定处理对HBV-DNA复制能力的影响。结果:测序鉴定rtM204I质粒构建成功。Southern杂交显示,突变型组HBV-DNA复制水平较野生型组降低;在拉米夫定灌胃处理后,野生型组HBV-DNA复制中间体水平较空白对照组明显下降(P<0.05),但突变型组HBV-DNA复制中间体水平与空白对照组比较,差异无统计学意义(P>0.05)。结论:乙肝病毒多聚酶区(P区)rtM204I位点突变降低HBV-DNA复制能力,并影响拉米夫定对HBV-DNA复制的抑制作用。

Objective:To establish a mouse model of hepatitis B virus YMDD mutation and study its replication capacity of HBV-DNA in vivo.Methods:A plasmid with hepatitis B virus polymerase region(P region)rtM204I mutation was established,and the mutant plasmid was injected into the mice through tail vein high pressure to detect the replication capacity of hepatitis B virus in the liver tissues of mice and the changes in the replication of lamivudine.Results:The plasmid of rtm204I was successfully constructed by sequencing identification,and the replication capacity of the plasmid in mice was lower than that of wild-type mice,but the replication capacity of HBV-DNA in mice injected with the mutant plasmid was significantly lower than that of wild-type mice.Conclusion:The mutation of rtM204I in hepatitis B virus polymerase region(P region)will reduce the replication capacity of HBV-DNA and decrease the susceptibility to lamivudine.