目的：研究一枝蒿黄酮类提取物抗鸭乙型肝炎病毒（DHBV）作用及化学成分。方法一枝蒿药材经50％乙醇提取，采用聚酰胺柱进行分离纯化，得到一枝蒿30％、50％乙醇洗脱物。采用1日龄北京鸭，人工感染鸭乙型肝炎病毒模型，将阳性鸭随机分为30％乙醇洗脱物组、50％乙醇洗脱物组、阿德福韦酯组、病毒对照组，各组分别于给药前、给药期间不同时间（5 d和10 d）和停药后3 d取血，分离血清；停药后第3天杀剖，取肝脏，以血清DH-BV-抗原、血清DHBV-DNA、肝脏 DHBV-DNA 作为药效评价指标，研究其体内抗乙肝病毒的作用。采用 LC-HRMS／MS法对有效部位化学成分进行分析。结果50％乙醇洗脱物在200 mg／kg剂量下能显著抑制鸭乙肝病毒血清和肝脏中DNA复制，30％乙醇洗脱物的抑制作用较弱。从50％乙醇洗脱物中推测出5种主要化合物，分别为6-去甲氧基-4′-O-甲基茵陈色原酮-7-O-β-D-葡萄糖苷、紫花牡荆素、异山奈甲黄素、洋艾素、山柰素-3，3′，4′-三甲醚。结论50％乙醇洗脱物有抗 HBV的作用，50％乙醇洗脱物中推测出的5种主要化合物可能在抗 HBV中起到重要的作用。%Objective To study the anti-DHBV effect and chemical constituents of flavonoids extract from Artemisia rupestris .Methods The separation and purification of the 50% ethanol extract of Artemisia rupestris L.were performed with polyamide column.30% and 50% ethanol-eluted fractions were obtained. 1-day old Beijing ducklings were used to establish the model of duck Hepatitis B Virus.The positive ducks were randomly divided into 30% ethanol extract group,50% ethanol extract group,adefovir dipivoxil group and virus control group.The blood of ducks were collected and the serum were isolated before medi-cation,and at the 5th,10th day after medication,and the 3rd day after stopping medication;These ducks were killed and livers were removed on the third day after stopping medication.The activity of in vivo anti-HBV was determined by Serum DHBV antigen,Serum DHBV-DNA and Liver DHBV-DNA detection with the method of immunohistochemistry.The ethanol-eluted fractions and the components were analyzed by LC-HRMS/MS.Results 50% ethanol-eluted fraction can significantly inhibit DHBV-DNA replication in serum and liver in the 200mg/kg dose.30% ethanol-eluted fraction has weak inhibition.Five compounds were gained from 50% ethanol-eluted fraction as 6-demethoxy-4′-O-methylcapillarisin-7-O-β-D-glucopyr-anoside,Caticin,kaempferide,artemetin,kaempferol 3,3′,4′-trimethylether.Conclusion 50% ethanol-e-luted fraction can significantly inhibit DHBV-DNA replication in serum and liver,indicating that 50% eth-anol-eluted fraction has an effect on anti-HBV.
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