霉酚酸对乙型肝炎病毒复制的体外实验观察

In vitro antiviral efficacy of mycophenolic acid against hepatitis B virus

目的探讨霉酚酸的体外抗乙型肝炎病毒作用.方法以HepG2.2.15为细胞模型,加入不同剂量的霉酚酸或拉米夫定,或霉酚酸加拉米夫定共培养,于第3、6、9天收集培养上清液行HBsAg定量检测;收集细胞提取总RNA,行逆转录聚合酶链反应(R-PCR)及荧光实时定量PCR测细胞内HBV mRNA水平.结果霉酚酸单独应用时,浓度2～250μg/ml无明显抗乙型肝炎病毒作用,当浓度达1250μg/ml时,可使病毒mRNA水平下降1～2 log,上清液中HBsAg滴度也有相应降低;拉米夫定5μg/ml可使病毒mRNA水平下降3～4 log,上清液中HBsAg滴度也有相应降低;霉酚酸和拉米夫定合用时,可显著增强后者的抗乙型肝炎病毒作用,其中浓度10μg/m1时效果最明显.结论霉酚酸有体外抗乙型肝炎病毒作用,并可增强拉米夫定的体外抗乙型肝炎病毒作用.

Objective To evaluate the in vitro inhibitive activities of mycophenolic acid against hepatitis B virus in vitro. Methods The different concentrations of the test drugs in DMEM were added to the well with the monolayer growth of HepG2.2. 15 cells. The culture medium was refreshed with DMEM containing the appropriate drugs every3 days. At 3rd, 6thandgth days, the supernatant was collected for HBsAg quantitative assay, total RNA was isolated from cells for RT PCR and real time-PCR assays. Results It was found no significant anti-HBV effect of mycophenolic acid on con centration of 2-250 μg/ml, and mini inhibitory efficacy of HBV replication on concentration of 1250μg/ml. There was significant anti HBV effect of lamivudine on concentration of 5 μg/ml. When combined with lamivudine mycophenolic acid can significantly potentiate anti HBV activities of lamivudine, and it can get maximal inhibition effect on concentration of 10μg/ml. Conclusions Mycophenolic acid could inhibit HBV replication and potentiate anti HBV activities of lamivudine.