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目的观察小鼠体内乙型肝炎病毒(HBV)X蛋白(HBx)对HBV基因转录和复制水平的影响。方法应用高压注射体内转染法将野生型HBV重组质粒、HBx表达缺失的HBV重组质粒及HBx表达质粒DNA通过尾静脉快速注射入小鼠体内,应用Southern和Northern印迹分别检测小鼠肝组织中HBV DNA复制中间体及HBV mRNA水平。结果野生型及HBx表达缺失的HBV重组质粒注射组小鼠肝组织中均检测到HBV的复制与转录;HBx表达缺失的HBV重组质粒注射组HBV基因转录和复制水平较野生型HBV重组质粒注射组、HBx表达缺失HBV重组质粒与HBx表达质粒共注射组弱,生理盐水注射组未检测到HBV的转录与复制。结论HBx的表达缺失可导致体内HBV基因转录和复制水平的下降,而外源HBx的表达可逆转这种因HBx表达缺失引起的HBV复制及转录的减弱,提示HBx增强小鼠体内HBV基因转录和复制。
Objective To observe the effect of hepatitis B virus (HBV) X protein (HBx) on HBV gene transcription and replication in mice. Methods The wild-type HBV recombinant plasmid, HBV recombinant plasmid with HBx expression loss and HBx expression plasmid DNA were rapidly injected into the tail vein through high-pressure injection in vivo. Southern and Northern blot were used to detect the expression of HBV DNA replication intermediates and HBV mRNA levels. Results HBV replication and transcription were detected in the liver of mice injected with HBV recombinant plasmids with wild-type and HBx-deficient cells. Compared with the wild-type HBV recombinant plasmids injected group, the level of HBV gene transcription and replication in HBx- , The expression of HBx was absent in HBV co-injection group and the HBx expression plasmid co-injection group was weak. No transcription and replication of HBV was detected in saline injection group. Conclusions The loss of HBx expression leads to the decrease of HBV gene transcription and replication in vivo. However, the expression of HBx can reverse the HBV replication and transcription decrease induced by the loss of HBx expression, suggesting that HBx enhances HBV gene transcription and copy.