基于网络药理学和细胞实验探究盐酸药根碱抗酒精性肝炎的作用机制  

Mechanism of jatrorrhizine hydrochloride in anti-alcoholic hepatitis based on network pharmacology and cellular experiments

作　　者：陈心雨 赵冰洁 郑浩东 贾爱亭 张志君 CHEN Xinyu;ZHAO Bingjie;ZHENG Haodong;JIA Aiting;ZHANG Zhijun(School of Pharmacy,Hubei University of Science and Technology,Xianning 437100,Hubei Province,China)

机构地区：[1]湖北科技学院药学院,湖北咸宁437100

出　　处：《数理医药学杂志》2024年第2期91-99,共9页Journal of Mathematical Medicine

摘　　要：目的基于网络药理学技术和体外细胞实验探究盐酸药根碱(jatrorrhizine hydrochloride,JH)治疗酒精性肝炎(alcoholic hepatitis,AH)的作用机制,为后续研究提供参考。方法基于PubMed数据库查询JH药物成分,借助SWISS数据库预测药物作用目标,在GeneCards和OMIM数据库中找出AH的目标点。通过DAVID数据库实现基因本体(gene ontology,GO)分析和京都基因与基因组百科全书(Kyoto Encyclopedia of Genes and Genomes,KEGG)富集分析,利用STRING数据库和Cytoscape软件建立蛋白质互作网络(protein-protein interaction network,PPI),并通过体外细胞实验验证。利用脂多糖(lipopolysaccharide,LPS)诱导RAW264.7细胞构建体外炎症模型,将细胞分为五组,采用MTT法测定不同浓度JH对细胞活性的作用,采用酶联免疫吸附测定法(enzyme-linked immunosorbent assay,ELISA)测定JH对肿瘤坏死因子-α(tumor necrosis factor-α,TNF-α)、白细胞介素-6(interleukin-6,IL-6)等炎症因子产生和释放的影响。结果利用网络药理学分析得到25个JH与AH的关键靶点,结合GO和KEGG富集分析及PPI网络构建结果,确定ESRI、NT5E、PGR等基因为后续实验靶点。根据ELISA实验结果进一步证实,相较于对照组,LPS组RAW264.7的M1型巨噬细胞中与炎症相关的因子TNF-α和IL-6的分泌显著增加;相较于LPS组,含有JH的组别细胞上清液中炎症因子TNF-α和IL-6的分泌有降低的趋势,尤其是随着JH浓度增加,其降低效果更加显著。结论在炎症模型细胞中,JH可以调控M1极化,表明JH可能是治疗AH的潜在分子。Objective To explore the mechanism of jatrorrhizine hydrochloride(JH)in the treatment of alcoholic hepatitis(AH)based on network pharmacology and in vitro cellular experiments.Methods Drug ingredients of JH were queried based on PubMed,drug action targets were predicted based on SWISS database,and AH target points were found in GeneCards and OMIM databases.Gene ontology(GO)and Kyoto Encyclopedia of Genes and Genomes(KEGG)enrichment analysis were performed based on DAVID database.The protein-protein interaction network(PPI)was established by using STRING database and Cytoscape software,and verified by in vitro cell experiments.An inflammation model was developed in vitro using RAW264.7 cells stimulated with lipopolysaccharide(LPS),and these cells were then categorized into five groups.The effect of different concentrations of JH on cell activity was determined by MTT assay.Enzyme-linked immunosorbent assay(ELISA)was used to determine the activity of JH on the production and release of inflammatory factors,such as tumor necrosis factor-α(TNF-α)and interleukin-6(IL-6).Results The network pharmacology yielded a total of 25 key targets of JH and AH.Combined with the results of GO and KEGG enrichment analysis and PPI construction,ESRI,NT5E,and PGR and other genes were identified as the targets for subsequent experiments.The results of ELISA further confirmed that the secretion of inflammation-related factors TNF-αand IL-6 in M1 macrophages of RAW264.7 in LPS group was significantly increased compared with that in the control group.Compared with LPS group,the secretion of inflammatory factors TNF-αand IL-6 in the cell supernatant of the groups containing JH had a tendency to decrease,especially with the increase of JH concentration,the decrease effect was more significant.Conclusion In inflammatory model cells,JH can regulate M1 polarization,suggesting that JH is a potential molecule for the treatment of AH.

关 键 词：酒精性肝炎 盐酸药根碱 抗炎机制 网络药理学 细胞实验 分子对接 

分 类 号：R285.5[医药卫生—中药学]