恩替卡韦和干扰素序贯作用对 HepG2.2.15细胞HBV DNA复制的影响

摘要

Objective To investigate the effect on replication of HBV DAN in HepG2 . 2 . 15 cells by the sequential treatment with Entecavir (ETV) following interferon α-2b (IFNα-2b). Methods In individual groups, HepG2. 2. 15 cells were treated with various concentrations of ETV (1. 5 μg/ml, 5 μg/ml, 10 μg/ml) and IFNα-2b (5 000 IU/ml, 10 000 IU/ml, 15 000 IU/ml) individually; in combination group, HepG2. 2. 15 cells were treated with ETV and IFNα-2b combination;in sequential groups, HepG 2. 2. 15 cells were treated with ETV following IFNα-2b. The method of determined proliferation was MTT; HBV DNA copies were detected by Real time PCR-fluorescence probing in one-tube method from the supermatant of HepG 2 . 2 . 15 cells; HBsAg and HBeAg expressions from the supermatant were measured by ELISA at different time points. Results Compared with the individual groups or the combination group, sequential group had more powerful inhibition effects on HBV DNA replication, the more powerful inhibition expressions of HBsAg and HBeAg in HepG2. 2. 15 cells. Conclusion The sequential treatment with ETV following IFNα-2b has more powerful effect on replication of HBV DNA than the individual treatment or combination treatment.%目的：探讨恩替卡韦(Entecavir,ETV)与干扰素(IFNα-2b)序贯作用对HepG2.2.15细胞HBV DNA复制的影响。方法将培养HepG2.2.15细胞分组,单独组中分别加入不同浓度ETV的配制液(1.5μg/ml、5μg/ml、10μg/ml)或 IFNα-2b配制液(5000 IU/ml、10000 IU/ml、15000 IU/ml)；联合组中联合加入ETV和IFNα-2b配制液；序贯组中先后加入ETV和IFNα-2b序贯配制液。在不同时点,采用MTT法检测HepG2.2.15细胞增殖情况,采用实时荧光定量法检测各组细胞培养上清液中HBV DNA水平,酶联免疫法(ELISA)检测上清液中HBsAg与HBeAg水平。结果随着时间延长,序贯组对HepG2.2.15的HBV DNA复制抑制作用明显大于其他两组(P<0.05)；对HBsAg与HBeAg的抑制作用随时间的延长而加强(P<0.05)。结论 ETV与IFNα-2b序贯作用于HepG2.2.15细胞,对HepG2.2.15细胞HBV DNA复制有明显的抑制作用,且作用强于单独或联合使用。