摘要
目的:采用网络药理学分析方法研究荔枝叶的抗炎作用机制,并进行体外细胞实验验证。方法:通过查阅文献查找荔枝叶化学成分,借助中药系统药理学数据库与分析平台(TCMSP)及SwissADME五项原则筛选出活性成分;基于Pubchem数据库、Swiss Target Prediction数据库预测各成分靶点;并利用Disgenet数据库获取炎症基因;采用Venny2.1.0数据库获得成分靶点与炎症基因的交集靶点作为潜在抗炎靶点,并将其输入STRING 11.0数据库获取蛋白质-蛋白质相互作用(PPI)网络图;使用DAVID 6.8数据库对抗炎靶点基因进行基因本体(GO)富集分析和京都基因和基因组百科全书(KEGG)通路富集分析;采用Cytoscape 3.8.0软件构建荔枝叶“活性成分-抗炎靶点-通路”网络图。通过CCK-8法检测荔枝叶醇提物对RAW264.7细胞活性的影响;酶联免疫吸附试验法检测肿瘤坏死因子-α(TNF-α)、白细胞介素-6(IL-6)的含量;蛋白质印迹检测环氧合酶-2(COX-2)蛋白表达。结果:从荔枝叶中筛选到9个活性成分,潜在抗炎靶点47个,127种GO生物功能,68条KEGG信号通路,涉及肿瘤坏死因子(TNF)信号通路、缺氧诱导因子1(HIF-1)信号通路等与炎症相关通路。细胞实验表明荔枝叶醇提物浓度为12.5,25,50μg/mL对小鼠腹腔巨噬细胞(RAW264.7)没有毒性,可降低脂多糖诱导RAW264.7细胞导致的TNF-α、IL-6的释放量,对COX-2蛋白的表达也有抑制作用。结论:构建的“成分-靶点-通路”网络结构图揭示了荔枝叶通过多成分、多靶点、多通路协同作用达到抗炎作用,为进一步阐明荔枝叶抗炎作用机制提供理论依据。
Objective:The anti-inflammatory mechanism of the leaves of Litchi chinensis Sonn.was studied by network pharmacological analysis and verified by in vitro cell experiments.Methods:The chemical constituents of the leaves of Litchi chinensis Sonn.were searched by literature.The active ingredients of the leaves of Litchi chinensis Sonn.were screened out by a traditional Chinese medicine systems pharmacology database and analysis platform(TCMSP)and the five principles of SwissADME.Each component target was predicted based on Pubchem database and Swiss target prediction database;inflammation genes were obtained from Disgenet database.The intersection target of the component target and inflammatory gene was obtained by Venny2.1.0 database as a potential anti-inflammatory target,and the protein-protein interaction(PPI)network diagram was constructed by inputting the target into STRING 11.0 database.Gene ontology(GO)enrichment analysis and Kyoto encyclopedia of genes and genomes(KEGG)pathway enrichment analysis of anti-inflammatory target genes were conducted by using DAVID 6.8 database.Cytoscape 3.8.0 software was used to construct the network diagram of active ingredients-anti-inflammatory targets-pathways of the leaves of Litchi chinensis Sonn.The effect of alcohol extract of the leaves of Litchi chinensis Sonn.on the activity of RAW264.7 cells was detected by CCK-8 method.The contents of tumor necrosis factor-α(TNF-α)and interleukin-6(IL-6)were detected by enzyme-linked immunosorbent assay(ELISA);the protein expression of cyclooxygenase-2(COX-2)was detected by Western blot.Results:Nine active ingredients were screened from the leaves of Litchi chinensis Sonn.,including 47 potential anti-inflammatory targets,127 GO biological functions,and 68 KEGG signal pathways,involving TNF signal pathway,hypoxia-inducible factor 1(HIF-1)signal pathway,and other inflammation-related pathways.The cell experiments showed that the alcohol extract concentrations of the leaves of Litchi chinensis Sonn.of 12.5,25,and 50μg/mL displayed no significant cytotoxicity in mouse peritoneal macrophage(RAW264.7).The concentration could reduce the release of TNF-αand IL-6 generated by RAW264.7 cells induced by lipopolysaccharide and inhibit the expression of COX-2 protein.Conclusion:The constructed network diagram of components-targets-pathways reveals that the leaves of Litchi chinensis Sonn.exert anti-inflammatory effects through joint efforts of multi-components,multi-targets,and multi-pathways.The results can provide a theoretical basis for further elucidating the anti-inflammatory mechanism of the leaves of Litchi chinensis Sonn.
作者
祁金丽
梁洁
黄冬芳
陈辉华
韦金玉
胡珏
刘星晨
黄团心
QI Jinli;LIANG Jie;HUANG Dongfang;CHEN Huihua;WEI Jinyu;HU Jue;LIU Xingchen;HUANG Tuanxin(Guangxi University of Chinese Medicine,Nanning 530200,China;Guangxi Zhuang Yao Medicine Center of Engineering and Technology,Nanning 530200,China;Guangxi Superior Chinese Patent Medicine and National Medicine Development Engineering Technology Center,Nanning 530200,China;South China Branch of National Engineering Research Center for Manufacturing Technology of Traditional Chinese Medicine Solid Preparation,Nanning 530200,China;Guangxi Institute for Food and Drug Control,Nanning 530021,China)
出处
《世界中医药》
CAS
2023年第8期1064-1071,共8页
World Chinese Medicine
基金
广西创新驱动发展专项资金项目(桂科AA17202046)
广西高等学校千名中青年骨干教师培育计划项目(桂教人〔2019〕5号)
中药学广西一流学科项目(桂教科研〔2018〕12号)
壮瑶药协同创新中心项目(桂教科研〔2013〕20号)
广西壮瑶药重点实验室建设项目(桂科基字〔2014〕32号)
广西第八批自治区特聘专家聘任人员项目(桂人才通字〔2019〕13号)
广西高等学校高水平创新团队及卓越学者计划项目(桂教师范〔2019〕52号)
广西科技基地和人才专项(桂科AD20238058)。
关键词
荔枝叶
抗炎
网络药理学
体外细胞实验
中药
活性成分
靶点
通路
Leaves of Litchi chinensis Sonn.
Anti-inflammation
Network pharmacology
In vitro cell experiment
Chinese traditional medicine
Active ingredients
Target
Pathway
