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目的 通过观测人ＨＢＶ ＤＮＡ在非哺乳动物──鸭肝细胞中的复制和表达水平，探讨人ＨＢＶ感染与复制的跨种属特异性，为建立人ＨＢＶ ＤＮＡ转染跨种属肝细胞模型奠定基础。方法 获取线性ＨＢＶ ＤＮＡ并电转染原代鸭肝细胞，电转后４８ｈ采用ＩＭＸ系统检测鸭肝细胞中ＨＢｓＡｇ和 ＨＢｅＡｇ表达水平，用 Ｓｏｕｔｈｅｒｎ ｂｌｏｔ－ｔｉｎｇ和ｄｏｔ ｂ１ｏｔｔｉｎｇ检测ＨＢＶ ＤＮＡ复制情况。以单纯电击肝细胞为对照。结果 转染组原代鸭肝细胞裂解液中ＨＢｓＡｇ为９．１０（Ｐ／Ｎ值≥ ２．１为阳性），ＨＢｅＡｇ为阴性；上清液中二者均为阴性。转染组原代鸭肝细胞裂解液ｄｏｔｂｌｏｔｔｉｎｇ呈强阳性；转染组肝细胞总 ＤＮＡ Ｓｏｕｔｈｅｒｎ ｂｌｏｔｔｉｎｇ显示约４．０ ｋｂ以下分子涂抹带，为游离复制型 ＨＢＶＤＮＡ，包括 ｒｃＤＮＡ、ｃｃｃＤＮＡ与 ｓｓＤＮＡ等复制中间体；未见整合型 ＨＢＶ ＤＮＡ──高分子区（４．０－２４．０ ｋｂ）涂抹带。对照组上述指标均为阴性。结论 人ＨＢＶ ＤＮＡ能在原代鸭肝细胞中复制和表达，可能为肝细胞内环境依赖性，无严格种属特异性限制。
OBJECTIVE: To investigate the cross-species specificity of human HBV infection and replication by observing the replication and expression of human HBV DNA in non-mammalian duck hepatocytes and lay the foundation for the establishment of a trans-species hepatocyte model of human HBV DNA . METHODS: Linear HBV DNA was obtained and electroporated into primary duck hepatocytes. The expression of HBsAg and HBeAg in duck hepatocytes was detected by IMX system 48 hours after electrotransfection, and HBV DNA replication was detected by Southern blotting and dot blotting. A simple shock of hepatocytes as a control. Results HBsAg in primary duck hepatocyte lysate was 9.10 (P / N value≥2.1 positive) and negative for HBeAg in the transfection group. Both of them were negative in the supernatant. Transfection group of primary duck hepatocyte lysate dot blotting was strongly positive; Transfection group of total DNA Southern blotting showed about 4.0 kb molecular smear zone, free replication type HBVDNA, including rcDNA, cccDNA and ssDNA replication intermediate Body; no integrated HBV DNA ─ ─ polymer area (4.0-24.0 kb) smear zone. The above indicators in the control group were negative. Conclusions Human HBV DNA can be replicated and expressed in primary duck hepatocytes, which may be dependent on the intracellular environment of hepatocytes without strict species-specific restriction.