女贞子抗丙型肝炎病毒复制酶活性成分研究  

Active components of Ligustrum lucidum inhibiting hepatitis C virus replicase activity

作　　者：孙瑞娜[1] 张艳妮[1] 汪俊[1] 刘好桔[1] 孔令保[1] 

机构地区：[1]江西农业大学生物科学与工程学院,江西南昌330045

出　　处：《药学学报》2013年第9期1390-1396,共7页Acta Pharmaceutica Sinica

基　　金：国家自然科学基金资助项目(31160034);江西省科技支撑计划重点项目(2009BNA09400);江西省自然科学基金资助项目(2010GZY0059)

摘　　要：在前期发现女贞子(Ligustrum lucidum,LL)提取物抑制丙型肝炎病毒(hepatitis C virus,HCV)复制酶(NS5B)活性的基础上,研究女贞子提取物抑制NS5B活性的化学成分及其作用机制。采用乙酸乙酯萃取和薄层色谱法分离女贞子水提取物,NS5B活性抑制实验检测各个分离组分的抑制活性。结果显示,分离组分1和2能够抑制NS5B活性,组分2的抑制能力更强。采用高效液相色谱法(high-performance liquid chromatography,HPLC)及NS5B活性抑制实验分析组分1和2中的活性成分。结果显示,熊果酸和齐墩果酸分别是组分1和2的抗NS5B活性成分,齐墩果酸的抑制能力更强。进一步的抑制机制分析发现,熊果酸和齐墩果酸通过非竞争抑制作用抑制NS5B活性,熊果酸和齐墩果酸的Ki分别约为4.7μg·mL 1(10μmol·kg-1)和2.5μg·mL-1(5.5μmol·kg-1)。本研究发现齐墩果酸和熊果酸通过非竞争抑制作用抑制丙型肝炎病毒复制酶NS5B活性,揭示这两个天然化合物具有潜在的丙型肝炎治疗价值。Based on previous report that the Chinese herb Ligustrum lucidum （LL） extract directly inhibited hepatitis C virus （HCV） replicase （NSSB） activity, the active components of LL extract to inhibit HCV NS5B activity and their inhibition mode were investigated in this study. LL extract was separated using ethyl acetate and thin layer chromatography （TLC）. The inhibitory activity of separated fractions on HCV NSSB was analyzed by the inhibitory assay of NS5B activity. The results showed that only fractions 1 and 2 inhibited NSSB activity, and fraction 2 possessed higher inhibitory activity than fraction 1. HPLC analysis combined with inhibitory assays indicated that ursolic acid and oleanolic acid are the active components within fractions 1 and 2 to inhibit NS5B activity, separately. Moreover, oleanolic acid possessed higher inhibitory activity than ursolic acid. Further inhibition mode analysis found that both oleanolic acid and ursolic acid suppressed NS5B activity as noncompetitive inhibitors. The Ki values of ursolic acid and oleanolic acid were about 4.7 μg·mL （10 μmol.kg^-1） and 2.5μg·mL 1 （5.5 μmol·kg^-1）, respectively. Taken together, these results demonstrated that oleanolic acid and ursolic acid suppressed NSSB activity as noncompetitive inhibitors, implying that the two natural products have potential value for HCV therapy.

关 键 词：丙型肝炎病毒 丙型肝炎病毒NS5B蛋白 女贞子 熊果酸 齐墩果酸 

分 类 号：R285[医药卫生—中药学]