氧化苦参碱在鸭原代肝细胞中抗鸭乙肝病毒(DHBV)作用的研究

通过建立鸭原代肝细胞-DHBV感染模型研究氧化苦参碱抗DHBV的作用。分别在DHBV感染前、感染同时以及感染后给药,利用打点杂交、Southern印迹核酸杂交和荧光定量PCR方法分别检测培养细胞上清及细胞内病毒核酸,观察氧化苦参碱在病毒感染的各个环节所起的抗病毒作用。实验结果显示:1mg/mL氧化苦参碱处理细胞后,鸭原代肝细胞培养上清及细胞内的DHBV核酸明显低于病毒感染对照组,病毒抑制率达91.6%;在病毒感染同时加药对病毒的抑制率可达98.5%;感染后持续用药能使不同培养天数的鸭肝细胞内的DHBV核酸降低60.5%~96.6%;氧化苦参碱与DHBV共孵育后,可以使病毒感染力下降69.6%。结果说明氧化苦参碱可以在DHBV感染鸭原代肝细胞的多个环节,包括病毒吸附、进入细胞及细胞内复制等方面发挥抗病毒作用。

Study of Anti-Duck Hepatitis B Virus Effects of Oxymatrine Using Primary Duck Hepatocyte Culture System

A primary duck hepatocyte-DHBV(PDH-DHBV) infection model was established to investigate the antiviral effects of Oxymatrine(OM).OM was added into primary duck hepatocyte(PDH) cultures at different steps of DHBV infection,namely,at the same time with DHBV challenge,and before or after PDH infection with DHBV.DHBV DNA in the culture supernatant was detected by dot blot hybridization analysis while intracellular viral DNA was detected by Southern blot hybridization analysis and the real-time PCR.Results showed that at the concentration of 1mg/mL,OM significantly suppressed viral DNA both in culture supernatant and in PDHs.The DHBV DNA inhibition efficiency in OM pretreated group was 91.6%;in the group where OM was added simutaneously with DHBV,the inhibitory efficiency was 98.5%;DHBV DNA replication was inhibited by 60.5%-96.6% at different days with continuous OM treatment postinfection and pretreating virus with OM made the viral infectivity decreased 69.6%.From these results,we conclude that OM has antiviral effects at the different stages of DHBV life cycle,including viral attachment,entry and replication in the host cells.