摘要
目的探讨江南卷柏治疗喉癌的分子机制。方法根据文献报道获取江南卷柏化学成分,SwissADME数据库筛选活性成分,PharmMapper数据库筛选作用靶点。检索OMIM等数据库收集喉癌相关靶点,Venny 2.1.0在线平台将二者取交集,STRNG平台进行蛋白相互作用分析。通过DAVID数据库进行GO和KEGG富集分析,使用Cytoscape 3.8.0软件构建可视化网络。通过SYBYL-X 2.0软件进行分子对接验证。利用MTT法、Hoechst 33258染色法、Western blot进行验证。结果在分子水平上,筛选出江南卷柏活性成分110个及药物靶点82个,喉癌相关靶点1642个,交集靶点34个。GO分析得到135个条目,KEGG分析共得到88条通路。分子对接结果显示,2″,3″-二氢金连木黄酮等11个关键活性成分和MAPK1等4个核心靶蛋白有95.5%有较好的对接活性。在细胞水平上,通过MTT法细胞活力测定筛选出SM-BFRE对喉癌细胞增殖的抑制作用最强,进一步通过Hoechst 33258染色表明SM-BFRE处理后的Hep-2细胞活力下降与细胞凋亡有关,最后通过Western blot验证SM-BFRE通过抑制PI3K/Akt/NF-κB/Cox-2通路来诱导喉癌细胞发生凋亡。结论充分体现了江南卷柏治疗喉癌多成分、多靶点、多通路协同作用的特点,为深入阐明江南卷柏治疗喉癌的作用机制提供理论参考。
Aim To explore the molecular mechanism of Selaginella moelledorffii Hieron.in the treatment of laryngeal cancer.Methods According to the relevant literature reports,the chemical constituents of S.moellendorffii were obtained,and the active ingredients were screened out through the SwissADME database,and the targets were screened through the PharmMapper database.The laryngeal cancer-related targets were collected by searching OMIM and other databases,and the Venny 2.1.0 online platform was used to obtain the intersection of the two.Protein interaction analysis of the potential targets was performed using the STRNG platform.GO functional analysis and KEGG pathway analysis was carried out using DAVID database.Visual networks were built with Cytoscape 3.8.0 software.Molecular docking was validated by SYBYL-X 2.0 software.MTT method,Hoechst 33258 staining method and Western blotting were also used for validation.Results At the molecular level,a total of 110 active ingredients of S.moellendorffii and 82 drug targets were screened out,1,608 targets related to laryngeal cancer,and intersection of 34 targets.GO analysis yielded 135 entries,and KEGG analysis yielded a total of 61 pathways.Molecular docking results showed that the 11 key active ingredients such as 2″,3″-dihydroochnaflavone wood flavonoids and 4 core target proteins such as MAPK1 had 95.5%of good docking activity.At the cellular level,SM-BFRE was screened for its strongest inhibitory effect on laryngeal cancer cell proliferation through MTT assay.Furthermore,Hoechst 33258 staining showed that the decrease in Hep-2 cell viability produced by SM-BFRE was related to cell apoptosis.Finally,Western blot verified that SM-BFRE inhibited PI3K/Akt/NF through inhibition-κB/Cox-2 pathway to induce apoptosis in laryngeal cancer cells.Conclusions To sum up,it fully reflects the multi-component,multi-target,and multi-channel synergistic effect of S.moellendorffii in the treatment of laryngeal cancer,and provides a theoretical reference for further elucidation of the mechanism of action of S.moellendorffii in the treatment of laryngeal cancer.
作者
李媛媛
王思斯
法缇玛·瑟菲迪肯
刘文琪
康丽
杨新洲
LI Yuan-yuan;WANG Si-si;Fatemeh Sefidkon;LIU Wen-qi;KANG Li;YANG Xin-zhou(College of Life Sciences,South-Central Minzu University,Wuhan 430074,China;College of Pharmaceutical Sciences,South-Central Minzu University,Wuhan 430074,China;Research Division of Medicinal Plants,Research Institute of Forests and Rangelands,Agricultural Research Education and Extension Organization(AREEO),Tehran,999067,Iran)
出处
《中国药理学通报》
CAS
CSCD
北大核心
2024年第2期352-362,共11页
Chinese Pharmacological Bulletin
基金
国家自然科学基金面上项目(No 81774000)
湖北省自然科学基金青年项目(No 2020CFB351)。
关键词
江南卷柏
喉癌
网络药理学
分子对接技术
体外细胞实验
作用机制
Selaginella moelledorffii Hieron
laryngeal cancer
network pharmacology
molecular docking technology
in vitro cellular assays
effect mechanism
