世界中医药

基金项目: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）

作者	单位
祁金丽1，梁洁1，2，3，4，黄冬芳1，陈辉华1，韦金玉1，胡珏1，刘星晨1，黄团心5	1 广西中医药大学药学院，南宁，530200； 2 广西壮瑶药工程技术研究中心，南宁，530200； 3 广西优势中成药与民族药开发工程技术研究中心，南宁，530200； 4 中药固体制剂制造技术国家工程研究中心华南分中心，南宁，530200； 5 广西壮族自治区食品药品检验所，南宁，530021

摘要点击次数: 603

全文下载次数: 0

中文摘要:

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12.5，25，50 μg/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。

English Summary:

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 50 μg/mL displayed no significant cytotoxicity in mouse peritoneal macrophage(RAW264.7).The concentration could reduce the release of TNF-α and 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.

查看全文查看/发表评论下载PDF阅读器