摘要
目的:基于体外实验和网络药理学方法探究淫羊藿(epimedii folium,EF)醇提物对乙型肝炎病毒(hepatitis B virus,HBV)的抑制作用及潜在机制。方法:使用EF醇提物处理野生型HBV稳定复制细胞系HepG 2.2.15细胞,采用CCK8法进行药物毒性评价;应用PCR-荧光探针“一管法”、ELISA法检测乙型肝炎病毒脱氧核糖核酸(HBV DNA)、乙型肝炎表面抗原(HBsAg)和乙型肝炎E抗原(HBeAg)水平,计算抑制率;利用TCMSP,GeneCards数据库筛选EF主要活性成分及HBV相关靶点,借助Cytoscape软件构建药物-活性成分-潜在疾病靶点网络,对核心靶点运用DAVID数据库进行GO和KEGG富集分析。结果:EF醇提物单独用药对HepG 2.2.15细胞的半数毒性浓度(CC_(50))值为1.21 mg·mL^(-1);对HBV DNA最大抑制率为59.54%,对HBsAg和HBeAg最大抑制率分别为63.07%和45.73%(P<0.05);对照药替诺福韦酯(tenofovir disoproxil fumarate,TDF)对HBsAg和HBeAg均未见明显抑制作用(抑制率<20.00%)。EF醇提物联合TDF对HBV DNA的最大抑制率为93.66%,对HBsAg和HBeAg的最大抑制率分别为54.97%和61.28%(P<0.05)。通过网络药理学分析共筛选出EF 23个候选活性成分及104个治疗HBV潜在靶点;GO富集分析得到624个条目,包括生物过程442个、细胞组分73个、分子功能109个;KEGG分析主要聚集在癌症通路、PI3K-Akt、脂质与动脉粥样硬化、人类乳头瘤病毒感染、乙型肝炎等信号通路。结论:EF可有效抑制野生型HBV复制,和TDF联合用药有协同抑制作用,尤其对抗原抑制效果显著;EF抗HBV具有多成分多靶点的作用特点,预测可以通过多条信号通路发挥作用,为后续深入探讨EF治疗HBV感染作用机制提供了理论基础。
Objective:To investigate the inhibitory effect and potential mechanism of Yinyanghuo(epimedii folium,EF)alcohol extract on hepatitis B virus(HBV) based on in vitro experiments and network pharmacology.Methods:The wild-type HBV stably replicating cell line HepG 2.2.15 cells were treated with EF alcohol extract and the drug toxicity was evaluated using CCK8 assay.The HBV DNA,HBsAg and HBeAg levels were detected by PCR-fluorescent probe “one-tube method” and ELISA,and the inhibition rate was calculated.The TCMSP and GeneCards databases were used to screen the main active ingredients of EF and HBV-related targets,and the drug-active ingredient-potential disease target network was constructed with Cytoscape software.GO and KEGG enrichment analyses were performed on the core targets using the DAVID database.Results:The CC_(50) value of EF alcohol extract alone on HepG 2.2.15 cells was 1.21 mg·mL^(-1),and the maximum inhibition rates for HBV DNA,HBsAg and HBeAg were 59.54%,63.07% and 45.73%,respectively(P<0.05).While the control drug tenofovir disoproxil fumarate(TDF) did not show significant inhibition for both HBsAg and HBeAg(inhibition rate 20.00%).The maximum inhibition rate of EF alcohol extract combined with TDF was 93.66% on HBV DNA,and the maximum inhibition rates on HBsAg and HBeAg were 54.97% and 61.28%,respectively(P<0.05).A total of 23 candidate active ingredients of EF and 104 potential targets for HBV treatment were screened by network pharmacology analysis.GO enrichment analysis yielded 624 entries,including 442 biological processes,73 cellular components and 109 molecular functions;and the KEGG analysis mainly focused on signaling pathways such as cancer pathway,PI3K-Akt,lipids and atherosclerosis,human papillomavirus infection and hepatitis B.Conclusion:EF can effectively inhibit the replication of wild-type HBV,and the combination of EF and TDF has a synergistic inhibitory effect,especially on antigen inhibition.EF has the characteristics of multi-component and multi-target action against HBV,which can be predicted to play roles through multiple signaling pathways.The findings provide a theoretical basis for subsequent in-depth exploration of the mechanism of EF in the treatment of HBV infection.
作者
郑艳芳
赵旭
曹梦珍
葛斐林
思兰兰
李乐
王伽伯
刘妍
肖小河
ZHENG Yan-fang;ZHAO Xu;CAO Meng-zhen;GE Fei-lin;SI Lan-lan;LI Le;WANG Jia-bo;LIU Yan;XIAO Xiao-he(College of Pharmacy,Fujian University of Traditional Chinese Medicine,Fuzhou 350122,China;Department of Hepatology,the Fifth Medical Center of Chinese PLA General Hospital,Beijing 100039,China;Department of Infectious Diseases,the Fifth Medical Center of Chinese PLA General Hospital,Beijing 100039,China)
出处
《中国新药杂志》
CAS
CSCD
北大核心
2023年第14期1458-1466,共9页
Chinese Journal of New Drugs
基金
国家自然科学基金重点资助项目(81930110)
国家自然科学基金创新群体资助项目(81721002)。
关键词
乙型肝炎病毒
淫羊藿醇提物
替诺福韦酯
抗病毒作用
网络药理学
hepatitis B virus
epimedii folium alcohol extract
tenofovir disoproxil fumarate
antiviral effect
network pharmacology
