耐干扰素的乙型肝炎病毒细胞模型的建立  

Construction of Interferon-Resistant HBV Cell Model

作　　者：韦佳佳 汤波[2] 徐丙发 秦侃 

机构地区：[1]安徽医科大学第三附属医院药学部,合肥230061 [2]安徽医科大学药学院,合肥230032

出　　处：《中国药学杂志》2017年第10期832-837,共6页Chinese Pharmaceutical Journal

基　　金：国家自然科学基金资助项目(81470165)

摘　　要：目的建立耐干扰素的乙型肝炎病毒(HBV)细胞模型,为进一步探索HBV对干扰素产生耐药机制提供实验基础。方法应用HBV体外转染细胞株HepG2.2.15,采用干扰素α-2b(IFNα-2b)低浓度(10~70 u·mL^(-1))长期诱导,历时48周建立HepG2.2.15/IFNα-2b细胞系模型。然后给予最佳药效浓度IFNα-2b培养4 d,比较低剂量诱导前后细胞上清液中HBsAg、HBeAg、HBV DNA的变化。结果经低浓度IFNα-2b刺激12周后,50 u·mL^(-1)组对最佳药效浓度IFNα-2b的敏感性显著降低,与刺激前细胞比较,HBsAg、HBeAg、HBV DNA的抑制率分别降低了25.48%、8.40%和15.43%,认为50 u·mL^(-1)为最佳刺激浓度。经50 u·mL^(-1)IFNα-2b分别刺激12~48周,发现36周后HBsAg、HBeAg、HBV DNA抑制率下降最明显,与刺激前细胞比较,抑制率分别降低了38.64%、15.71%和30.17%,认为36周为最佳刺激时间。结论经IFNα-2b持续诱导可使HepG2.2.15对IFNα-2b产生部分耐药,其中50 u·mL^(-1)IFNα-2b持续诱导36周最易使之产生耐药。OBJECTIVE To establish an interferon-resistant hepatitis B virus cell model and provide experimental basis for further investigating the mechanism of HBV resistance to interferon. METHODS HepG2. 2. 15 was continuously grown in the presence of 10,30,50 and 70 u·mL^-1 of interferon α-2b for up to 48 weeks. The HepG2. 2. 15/IFN α-2b cell model was constructed after 48 weeks of induction. The cells were treated with the best-effect concentration. Then, the levels of HBsAg, HBeAg, and HBV DNA in the supernatant of cell culture medium before and after the treatment were compared. RESULTS After stimulation with low concentrations of IFN α-2b for 12 weeks, the 50 u·mL^-1 group showed significant resistance to the best-effect concentration of IFN α-2b. Compared with the levels before stimulation, the inhibition rate on HBsAg, HBeAg, and HBV DNA decreased by 25.48% , 8.40% , and 15.43%, respectively, suggesting that 50 u·mL^-1 was the best-stimulation concentration. After stimulated with 50 u·mL^-1 IFNα-2b for 12 -48 weeks, the results showed that the inhibition rate on HBsAg, HBeAg, and HBV DNA after 36 weeks was the most significant. CONCLUSION Continuous induction with 50 u·mL^-1 IFN α-2b for 36 weeks could most easily induce drug resistance in HepG2. 2. 15 cells.

关 键 词：干扰素 乙型肝炎病毒 耐药 转染 细胞模型 

分 类 号：R978.7[医药卫生—药品] R965[医药卫生—药学]