基于网络药理学和蛋白验证技术探讨桔梗皂苷D通过调控Hippo信号通路对宫颈癌的作用机制  

Mechanism of platycodin D on cervical carcinoma by regulating Hippo signal pathway based on network pharmacology and targeted protein verification

作　　者：马琳琳[1] 杨新鸣[1] 时思毛[1] 郭滢[1] 韩凤娟[1] 杨阳[2] 李奇玮[3] MA Lin-lin;YANG Xin-ming;SHI Si-mao;GUO Ying;HAN Feng-juan;YANG Yang;LI Qi-wei(The First Affiliated Hospital of Heilongjiang University of Traditional Chinese Medicine,Harbin 150040;Harbin Medical University,Harbin 150040;Heilongjiang University of Traditional Chinese Medicine,Harbin 150040)

机构地区：[1]黑龙江中医药大学附属第一医院,哈尔滨150040 [2]哈尔滨医科大学,哈尔滨150040 [3]黑龙江中医药大学,哈尔滨150040

出　　处：《中南药学》2022年第12期2708-2714,共7页Central South Pharmacy

基　　金：国家自然科学基金青年科学基金项目(No.81702564);黑龙江省中医药科研项目(No.ZHY18-060);黑龙江中医药大学科研基金项目博士创新基金(No.LBH-Z19214)。

摘　　要：目的基于网络药理学、计算机模拟分子对接及蛋白靶向验证揭示桔梗皂苷D对宫颈癌的作用机制。方法从药物成分入手,基于反向对接及分子对接技术预测桔梗皂苷D在体内可能的作用靶标,同时从疾病入手,以“cervical carcinoma”为关键词汇总致病靶标。对两者结果取交集处理,获取共有的蛋白成分的潜在靶标成分;对潜在靶标进行体内作用关系分析(PPI)、基因本体(GO)富集分析和京都基因与基因组百科全书(KEGG)信号通路富集分析。构建成分-靶标-疾病的相互作用网络,通过分子对接软件模拟桔梗皂苷D与宫颈癌相关的靶蛋白的分子对接情况,最后以结合能、动物实验及分子生物学技术初步验证预测结果。结果桔梗皂苷D预测得到299个靶标,与宫颈癌相关的疾病靶标7140个,同时满足条件的靶蛋白48个,包括骨形态发生蛋白-2(BMP-2)等。GO分析显示桔梗皂苷D治疗宫颈癌可能涉及生物学途径288条、细胞组分11个、分子功能31条。KEGG信号通路富集筛选得到181条信号通路(P<0.05),其中主要涉及Hippo信号通路、癌症的途径、孕酮介导的卵母细胞成熟等。分子对接结果显示Hippo信号通路中BMP-2蛋白成分与桔梗皂苷D结合能稳定,小于-5 kcal·mol^(-1)。动物实验及Western结果与网络模拟结果基本一致。结论桔梗皂苷D可能通过对Hippo信号通路中BMP-2等蛋白的直接调控发挥药效,同时通过对癌症的途径、孕酮介导的卵母细胞成熟等过程等生物学过程的调控共同达到干预宫颈癌的目的。Objective To determine the mechanism of platycodin D on cervical carcinoma based on network pharmacology,molecular docking technology and targeted protein verification.Methods From the drug structure component,the potential targets of platycodin D in vivo were predicted based on reverse docking and molecular docking technology.While cervical carcinoma were used as the key word to sort out known targets with of diseases.The intersection of the two potential targets was obtained,and analyzed by PPI,GO enrichment and KEGG pathway enrichment.An interaction network with components,targets and diseases was built to simulate the molecular docking of platycodin D and targets related to cervical carcinoma through molecular docking software.Finally the verification was carried out with binding energy,animal experiments and molecular biology technology.Results Totally 299 potential proteins of platycodin D and 7140 disease targets related to cervical carcinoma were obtained.Totally 48 qualified targes,including BMP-2(bone morphogenetic protein-2),were obtained by reverse docking platform Pharmmapper.Go analysis showed that platycodin D involved 288 biological pathways,11 cellular components and 31 molecular functions in the treatment of cervical carcinoma.The enrichment and screening of KEGG pathway screened 181 signal pathways(P<0.05),mainly involved in the processes of Hippo signaling pathway,pathways in cancer and progesterone mediated oocyte maturation.Molecular docking showed that the binding energy between BMP-2 protein and platycodin D in Hippo signaling pathway was stable with binding energy less than-5 kcal·mol^(-1).The animal experiment and Western blot analysis showed basically consistent network simulation.Conclusion The mechanism of platycoside D in the treatment of cervical carcinoma may be directly related to the Hippo signaling pathway,which may exert its effect through the direct regulation of BMP-2 and other proteins in the Hippo signaling pathway.While intervention to cervical carcinoma can be achieved t

关 键 词：桔梗皂苷D 宫颈癌 网络药理学 信号通路 分子对接技术 

分 类 号：R285[医药卫生—中药学]