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目的:研究用丁型肝炎病毒(HDV)作为载体携带乙型肝炎病毒(HBV)特异性的锤头状核酶所构建的重组体,在细胞体系及转染动物模型中对HBV基因表达和复制的影响。方法:将HDV核酶重组体和HBV的共表达质粒转染Huh7细胞以分析HDV核酶重组体对HBV基因表达的影响;用小鼠尾静脉快速注射法将共表达质粒转染到小鼠体内,检测重组体在动物体内对HBV基因表达和复制的抑制作用。结果:转染细胞中,重组体对HBsAg的抑制与HDV重组位点和核酶靶位都有关;水压法注射的质粒在小鼠肝内得到表达,与对照相比重组HDV核酶可有效抑制在肝和血清中HBV的基因表达以及复制,与细胞中的结果一致。结论:此项体内实验为进一步构建治疗性重组HDV病毒,发现靶向性抗病毒基因治疗手段奠定基础。
OBJECTIVE: To study the expression of HBV gene and replication in a cellular and transfectant animal model using recombinant hepatitis B virus (HBV) specific hammerhead ribozyme carrying hepatitis B virus (HDV) as a vector Impact. Methods: The recombinant plasmid of HDV ribozyme and HBV was transfected into Huh7 cells to analyze the effect of HDV ribozyme recombination on HBV gene expression. Co-expression plasmid was transfected into mice by rapid tail vein injection , The inhibition of HBV gene expression and replication in animals was tested. Results: The inhibition of HBsAg in recombinant cells was related to both HDV recombination sites and ribozyme target sites. The plasmids injected by hydrostatic method were expressed in the liver of mice, and the recombinant HDV ribozymes could be effective compared with the control Inhibition of HBV gene expression and replication in liver and serum, consistent with the results in cells. Conclusion: This in vivo experiment laid the foundation for the further construction of therapeutic recombinant HDV virus and the discovery of targeted antiviral gene therapy.