乙型肝炎体外实验模型的方法学研究

Methodical Study on the Model of Hepatitis B in Vitro

目的本研究拟在原有体外模型,如体外鸭原代肝细胞培养模型、大鼠原代肝细胞培养和HBV基因转染人肝癌细胞系-2215细胞培养模型的基础上,对相关实验方法和内容进行改进,以便为防治乙肝药物的应用研究提供比较理想的实验方法。方法根据乙型肝炎的病症和乙肝病毒感染的特点,分别以嗜肝DNA病毒感染鸭的原代肝细胞、HBV基因转染人肝癌细胞系-2215细胞和CCl4熏染大鼠原代肝细胞体外培养试验,观察肝原代细胞培养后的存活状态、DHBVDNA和ALT的水平。结果受染后的鸭、2215细胞和鼠原代肝细胞存活时间较未染细胞明显缩短,DHBVDNA和ALT的水平显著升高。结论改进后的乙肝鸭原代肝细胞培养、CCl4熏染大鼠原代肝细胞培养和人肝癌2215细胞体外培养模型稳定,重复性好,适用于防治乙型肝炎有效药物的筛选和研究。

Aim To explore and improve many types of experimental models of hepatitis B in order to provide varied methods for the application study on drugs for prevention and cure of hepatitis B. Methods According to the disorder and characters of hepatitis B, was used the models of duck primary hepatoeytes which were infected with duck hepatitis B virus （DHBV）, the human hepatoeellular carcinoma （cell 2215, Hep G2） which was transferred with hepatitis B virus and rats primary hepatoeytes cultured with CC14 in vitro respectively. The length of survival and the levels of DHBV - DNA and ALT were observed in those models. Results The length of survival of infected - duck, cell 2215 and rats primary hepato- cytes were shorter than those of the control, and the level of DHBV - DNA and ALT increased significantly. Conclusion This method is stable in function and easily repeated and can be used to explore and set up many kinds of experimental models of hepatitis B in application study on drugs for prevention and cure of hepatitis B.