基于网络药理学和代谢组学的苦参乌梅汤对HepG 2.2.15细胞模型抗乙型肝炎病毒作用机制研究  

Study on the mechanism of Kushen Wumei Decoction against hepatitis B virus in HepG 2.2.15 cell model based on network pharmacology and metabolomics

作　　者：郑蕊[1] 郭朋[1] 刘宇[1] 孔伟[1] 陈艳[1] ZHENG Rui;GUO Peng;LIU Yu;KONG Wei;CHEN Yan(Department of Pharmacy,Xiyuan Hospital of China Academy of Chinese Medical Sciences,Beijing 100091,China)

机构地区：[1]中国中医科学院西苑医院药学部,北京100091

出　　处：《中国医药导报》2022年第15期12-18,共7页China Medical Herald

基　　金：国家自然科学基金面上项目(81673967)。

摘　　要：目的应用网络药理学和代谢组学技术探讨苦参乌梅汤对HepG 2.2.15细胞模型抗乙型肝炎病毒(HBV)的作用机制。方法采用酶联免疫吸附试验评价苦参乌梅汤对HepG 2.2.15细胞模型的药效影响。采用UHPLC-LTQ-Orbitrap-MS的代谢组学分析方法,结合主成分分析法及正交-偏最小二乘法分析法进行多元统计分析。另外,采用网络药理学的技术预测苦参乌梅汤抗HBV的潜在代谢通路。结合两部分获得的代谢通路,探究苦参乌梅汤抗HBV的作用机制。结果与空白组比较,1250μg/ml组和2500μg/ml组细胞存活率更低(P<0.05);与空白组比较,2500μg/ml组HBVDNA含量更低(P<0.05);筛选出组间具有显著性差异的代谢物40个,这些代谢物主要影响苯丙氨酸、酪氨酸和色氨酸生物合成、苯丙氨酸代谢、鞘脂代谢、精氨酸和脯氨酸代谢等;苦参乌梅汤候选活性成分的潜在治疗乙型肝炎靶点共115个,degree值排名前5位的靶点依次为蛋白激酶B、人体表皮生长因子受体、原癌基因酪氨酸蛋白激酶Src、肿瘤坏死因子和血管内皮生长因子A;基因本体富集分析到679个条目,包括生物过程2162个、细胞组分56个、分子功能133个;京都基因和基因组数据库通路富集分析,P值排名前20位的通路主要有PI3K-Akt、乙型肝炎、卡波西氏肉瘤相关疱疹病毒感染等信号通路。结论苦参乌梅汤可以抗HBV,代谢组学和网络药理学发现其作用的机制可能与调节鞘脂代谢通路、苯丙氨酸、酪氨酸和色氨酸的生物合成、苯丙氨酸代谢通路及PI3K-Akt信号通路有关,为抗HBV的作用机制提供了依据。Objective To explore the anti-hepatitis B virus(HBV)mechanism of Kushen Wumei Decoction on HepG 2.2.15 cell model by using network pharmacology and metabonomics technology.Methods Enzyme linked immunosorbent assay was used to evaluate the effect of Kushen Wumei Decoction on HepG 2.2.15 cell model.Metabolomics analysis method of UHPLC-LTQ-Orbitrap-MS was used,combined with principal component analysis method and orthogonal-partial least squares multiplicative analysis method were used for multivariate statistical analysis.In addition,the network pharmacology technology was used to predict the potential metabolic pathways of Kushen Wumei Decoction against HBV.Combining the metabolic pathways obtained from the two parts,the anti-HBV mechanism of Kushen Wumei Decoction was explored.Results Compared with blank group,the survival rates of 1250μg/ml and 2500μg/ml groups were lower(P<0.05);Compared with blank group,2500μg/ml group had lower HBVDNA content(P<0.05);40 metabolites with significant differences between groups were screened,which mainly affected the biosynthesis of phenylalanine,tyrosine,and tryptophan,phenylalanine metabolism,sphinolipid metabolism,arginine and proline metabolism;the potential therapeutic targets of the candidate active ingredients of Kushen Wumei Decoction were 115,and the top five targets in degree value were protein kinase B,human epidermal growth factor receptor,proto-oncogene tyrosine protein kinase Src,tumor necrosis factor,and vascular endothelial growth factor A.A total of 679 genes were identified by gene ontology enrichment analysis,including 2162 biological processes,56 cell components,and 133 molecular functions;Kyoto encyclopedia of genes and genomes pathway enrichment analysis showed that the top 20 P-value pathways mainly included PI3K-Akt,hepatitis B,Kaposi’s sarcoma associated herpes virus infection,and other signaling pathways.Conclusion Kushen Wumei Decoction can resist HBV.Metabolomics and network pharmacology found that the mechanism of its action may be related to t

关 键 词：苦参乌梅汤 抗乙型肝炎病毒 代谢组学 网络药理学 

分 类 号：R512.6[医药卫生—内科学]