黄根提取物抗乙型肝炎病毒活性的初步研究

摘要

目的:探讨黄根(Prismatomeris connata)提取物的抗乙型肝炎病毒(HBV)活性.方法:利用柱色谱、薄层色谱以及重结晶等技术对黄根提取物进行分离纯化,通过HPLC、FT-IR、1H NMR,13C NMR技术对化合物进行结构鉴定;选择HepG2.2.15细胞株为研究对象,以拉米夫定(3TC)作为阳性对照药物,MTT法检测药物对细胞的半数抑制浓度(IC50),酶联免疫吸附试验(ELISA)检测药物对细胞分泌HBeAg和HBsAg的影响.结果:从黄根乙酸乙酯提取部位中分离出纯度为95％以上的1,3-二羟基-2-甲基-9,10-蒽醌(甲基异茜草素);甲基异茜草素对HepG2.2.15细胞的IC50值为13.79 mg/L;甲基异茜草素对HepG2.2.15细胞的HBsAg和HBeAg均有一定的抑制作用且呈浓度依赖性;8 mg/L的甲基异茜草素对HepG2.2.15细胞HBeAg分泌作用强于3TC(100 mg/L) (P＜0.01).结论:甲基异茜草素抑制HepG2.2.15细胞分泌HBeAg作用明显,有望成为新型的抗HBV药物.%Objective:To study the effects of Prismatomeris Connata extracts on anti-hepatitis B virus.Methods:A compound was separated and purified from Prismatomeris Connata extracts by silica gel column,thin-layer chromatography and recrystallization.The structure of the compound was analyzed by HPLC,FT-IR,1H NMR and 13C NMR.Using lamivudine (3TC) as positive control drug,the half inhibitory concentration (IC50) of the purified compound on HepG2.2.15 cells was detected by MTT assay and its effect on HBeAg and HBsAg secretions was determined by enzyme-linked immunosorbent assay (ELISA).Results:The purity of the compound isolated from Prismatomneris Connata was greater than 95 ％ by HPLC analysis,and was identified to be 1,3-dihydroxy-2-methyl-9,10-anthraquinone (DMA).The IC50 of DMA on HepG2.2.15 cells was 13.79 mg/L and it remarkably decreased the amounts of secretion HBsAg and HBsAg in a dose manner.DMA (8 mg/L) inhibited HBeAg secretion more effectively than 3TC (100 mg/L) did (P＜0.01).Conclusion:DMA could reduce HBeAg secretion in HepG2.2.15 cell,and it might be a novel anti-HBV drug candidate.