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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 a model of Hepatitis B virus (HBV) resistant to interferon and to provide an experimental basis for further exploring the mechanism of HBV resistance to interferon. Methods HepG2.2.15 cells were transfected with HepG2.2.15 in vitro. HepG2.2.15 / IFNα was induced by interferon α-2b (10 ~ 70 u · mL -1) for 48 weeks. -2b cell line model. Then, the best concentration of IFNα-2b was cultured for 4 days. The changes of HBsAg, HBeAg and HBV DNA in the cell supernatant before and after low dose induction were compared. Results The sensitivity of 50 u · mL -1 group to IFNα-2b was significantly decreased after 12 weeks of stimulation with low concentration IFNα-2b. Compared with pre-stimulation cells, the sensitivity of HBsAg, HBeAg and HBV DNA The inhibitory rates were reduced by 25.48%, 8.40% and 15.43%, respectively. The optimal concentration was 50 u · mL -1. The inhibitory rates of HBsAg, HBeAg and HBV DNA decreased significantly after 36 weeks of treatment with 50 u · mL -1 IFNα-2b for 12-48 weeks, respectively. Compared with the pre-stimulation cells, the inhibitory rates were reduced by 38.64% 15.71% and 30.17%, think 36 weeks for the best time to stimulate. CONCLUSION: HepG2.2.15 is partially drug-resistant to IFNα-2b induced by IFNα-2b. 50 u · mL -1 IFNα-2b is most likely to induce drug resistance after 36 weeks of induction.