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目的观察融合蛋白胸腺素α1-干扰素α(TA1-IFN)体外抗乙型肝炎病毒(HBV)作用,并与胸腺素α1、干扰素α两者联合(TA1+IFN)应用的体外抗HBV作用进行比较。方法HepG22.2.15细胞接种后24h,换用含5种不同浓度(8000、4000、2000、1000、500U／ml)药物的培养基,37℃、体积分数5％CO2条件下培养,每3d换用原浓度含药培养液1次,并于第6天收集培养液,用Abbott诊断试剂盒,分别检测不同组药物作用后上清液中乙型肝炎表面抗原(HBsAg)、乙型肝炎e抗原(HBeAg)的含量,并计算其抑制率;同时用四甲基偶氮唑盐比色分析法检测不同组药物对HepG22.2.15细胞的细胞毒性作用。结果TA1-IFN体外对HBsAg、HBeAg的抑制率与药物浓度呈剂量依赖关系,并且在药物浓度达8000U／ml后趋于稳定,此时TA1-IFN对HBsAg、HBeAg抑制率分别为72.2％±0.8％、60.4％±1.1％,细胞存活率为85.2％±2.0％;而相应浓度的TA1-IFN对HBsAg、HBeAg抑制率为40.0％±0.7％、34.5％±3.2％,细胞存活率为70.0％±1.9％,两者HBsAg、HBeAg抑制率及细胞存活率比较差异均有统计学意义(P值均<0.05)。结论融合蛋白TA1-IFN体外具有良好的抗HBV作用,其体外抗HBV活性比TA1-IFN联合应用强,且细胞毒性比TA1+IFN联合应用时小,本实验为融合蛋白TA1-IFN的临床研究提供了重要的理论?
Objective To observe the in vitro anti-HBV effect of the fusion protein thymosin α1-interferon (TA1-IFN) against hepatitis B virus (HBV) in vitro and in combination with both thymosin α1 and interferon α (TA1 + IFN) Compare. Methods HepG22.2.15 cells 24h after inoculation, with 5 different concentrations (8000,4000,2000,1000,500 U / ml) of the medium, at 37 ℃, the volume fraction of 5% CO2 under the conditions of each change with The original concentration of drug-containing culture medium 1 time, and on the 6th day culture medium was collected, with Abbott diagnostic kit, were detected in different groups of drugs supernatant of hepatitis B surface antigen (HBsAg), hepatitis B e antigen HBeAg) were detected by enzyme-linked immunosorbent assay (ELISA). The inhibitory rate of HepG22.2.15 cells was calculated by MTT assay. The cytotoxicity of different groups of drugs on HepG22.2.15 cells was also detected by MTT colorimetric assay. Results The inhibitory rates of HBsAg and HBeAg in vitro were dose-dependent and dose-dependent when the concentration of TA1-IFN reached 8000U / ml. The inhibitory rates of TA1-IFN on HBsAg and HBeAg were 72.2% ± 0.8 %, 60.4% ± 1.1%, and cell survival rate was 85.2% ± 2.0%, while the corresponding concentrations of TA1-IFN inhibited HBsAg and HBeAg by 40.0% ± 0.7%, 34.5% ± 3.2% and cell survival rate was 70.0% ± 1.9%. The difference of HBsAg, HBeAg and cell survival between the two groups were statistically significant (P <0.05). Conclusion The fusion protein TA1-IFN has a good anti-HBV effect in vitro. Its in vitro anti-HBV activity is stronger than that of TA1-IFN and its cytotoxicity is less than that of TA1-IFN. This study is a clinical study of fusion protein TA1-IFN Provide important theory?