孟颖;蒋志涛;严国俊;沈晶;孙戡平;王媛媛;曹杰楠;夏梦莹;潘金火;
MENG Ying;JIANG Zhi-tao;YAN Guo-jun;SHEN Jing;SUN Kan-ping;WANG Yuan-yuan;CAO Jie-nan;XIA Meng-ying;PAN Jin-huo;Department of Pharmacy, Zhangjiagang Traditional Chinese Medicine Hospital Affiliated to Nanjing University of Chinese Medicine;College of Pharmacy, Nanjing University of Chinese Medicine;Department of Pharmacy, Wuxi Huishan Hospital of Traditional Chinese Medicine;Department of Pharmacy, Jinhu County People′s Hospital;

• 1:南京中医药大学附属张家港医院药学部
• 2:南京中医药大学药学院
• 3:江苏省无锡市惠山区中医医院药学部
• 4:江苏省金湖县人民医院药学部

摘要(Abstract)：

基于色谱-质谱和网络药理学-分子对接技术分析经典名方清胃散治疗牙周炎的作用机制。利用UPLC-Q-TOF-MS、GC-MS技术分析清胃散化学成分；经TCMSP数据库筛选清胃散活性成分及靶蛋白后利用SwissTargetPrediction数据库进行靶点预测；利用GeneCards、OMIM和DisGeNET数据库获取牙周炎相关治疗靶点；运用Venny 2.1构建韦恩图并得到交集靶点，运用Cytoscape 3.7.2构建“化合物-靶点-疾病”网络；利用clusterProfiler R功能包对潜在作用靶点进行GO功能和KEGG通路富集分析，构建“化合物-靶点-通路”网络，并利用分子对接技术验证活性成分与靶点蛋白的结合活性。通过色谱-质谱技术共分析得到清胃散中189种化学成分，筛选后得到39种活性成分对应180个潜在作用靶点与牙周炎相关，活性成分作用靶点富集分析得到92条KEGG通路，“化合物-靶点-通路”网络共涉及20条KEGG通路、34种活性成分和99个靶点，分子对接结果验证关键化合物与关键靶点有较好结合力。该研究初步表明，经典名方清胃散通过多成分、多靶点、多途径发挥治疗牙周炎的作用，体现了中药复杂系统的作用特点，为清胃散后续的物质基准及关键质量属性研究提供了理论依据。
The present study explored the mechanism of Qingwei Powder(QP) in the treatment of periodontitis based on chromatography-mass spectrometry and network pharmacology-molecular docking techniques. UPLC-Q-TOF-MS and GC-MS were used to identify the chemical constituents of QP. The active components and targets were screened out through the Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform(TCMSP), and their targets were predicted using SwissTargetPrediction. Targets related to periodontitis were obtained from GeneCards, OMIM, and DisGeNET. Venn diagram was constructed using Venny 2.1 to obtain the intersection targets. Cytoscape 3.7.2 was used to construct the "chemical component-target-disease" network. The targets were analyzed for Gene Ontology(GO) function and Kyoto Encyclopedia of Genes and Genomes(KEGG) pathway enrichment by clusterProfiler R, and the "chemical component-target-pathway" network was constructed. The binding activity of the active components to the target proteins was verified by molecular docking. A total of 189 chemical components were obtained by UPLC-Q-TOF-MS and GC-MS, including 39 active components with 180 potential targets related to periodontitis. Target enrichment analysis of the active components yielded 92 KEGG pathways. Twenty KEGG pathways, 34 active components, and 99 targets were involved in the "chemical component-target-pathway" network. Molecular docking verified a good binding ability of the key targets to the key compounds. This study preliminarily indicates that QP is effective in treating periodontitis through multiple components, multiple targets, and multiple pathways, which reflects the complex system of Chinese medicine. This study provides the theoretical foundation for the subsequent research on the material basis and key quality attributes of QP.

关键词(KeyWords)： 经典名方;清胃散;牙周炎;UPLC-Q-TOF-MS;GC-MS;网络药理学;分子对接
classical prescription;Qingwei Powder;periodontitis;UPLC-Q-TOF-MS;GC-MS;network pharmacology;molecular docking

Abstract:

Keywords:

基金项目(Foundation): 国家自然科学基金项目(81773910,82074004);; 国家重点研发计划项目(2019YFC1710603)

作者(Authors): 孟颖;蒋志涛;严国俊;沈晶;孙戡平;王媛媛;曹杰楠;夏梦莹;潘金火;
MENG Ying;JIANG Zhi-tao;YAN Guo-jun;SHEN Jing;SUN Kan-ping;WANG Yuan-yuan;CAO Jie-nan;XIA Meng-ying;PAN Jin-huo;Department of Pharmacy, Zhangjiagang Traditional Chinese Medicine Hospital Affiliated to Nanjing University of Chinese Medicine;College of Pharmacy, Nanjing University of Chinese Medicine;Department of Pharmacy, Wuxi Huishan Hospital of Traditional Chinese Medicine;Department of Pharmacy, Jinhu County People′s Hospital;

DOI: 10.19540/j.cnki.cjcmm.20211027.403

参考文献(References)：

• [1] KASSEBAUM N J,BERNABE E,DAHIYA M,et al.Global burden of severe periodontitis in 1990—2010:a systematic review and Meta-regression [J].J Dent Res,2014,93(11):1045.
• [2] PATINI R,SYADERINI E,LAJOLO C,et al.Relationship between oral microbiota and periodontal disease:a systematic review [J].Eur Rev Med Pharmacol Sci,2018,22(18):5775.
• [3] MOLON R S,ROSSA C J,THURLINGS R M,et al.Linkage of periodontitis and rheumatoid arthritis:current evidence and potential biological interactions [J].Int J Mol Sci,2019,20(18):4541.
• [4] GRAVES D T,CORREA J D,SILVA T A.The oral microbiota is modified by systemic diseases [J].J Dent Res,2019,98(2):148.
• [5] SANZ M,MARCO A,JEPSEN S,et al.Periodontitis and cardio-vascular diseases:consensus report [J].J Clin Periodontol,2020,47(3):268.
• [6] JIMENEZ M,KRALL E A,GARCIA R I,et al.Periodontitis and incidence of cerebrovascular disease in men [J].Ann Neurol,2009,66(4):505.
• [7] HOBBINS S,CHAPPLE I L,SAPEY E,et al.Is periodontitis a comorbidity of COPD or can associations be explained by shared risk factors/behaviors?[J].Int J Chron Obstruct Pulmon Dis,2017,12:1339.
• [8] WANG C J,MCCAULEY L K.Osteoporosis and periodontitis [J].Curr Osteoporos Rep,2016,14(6):284.
• [9] 李焕宝.阿莫西林联合甲硝唑治疗慢性牙周炎的效果探究[J].全科口腔医学电子杂志，2020,7(6):57.
• [10] GRAZIANI F,KARAPETSA D,ALONSO B,et al.Nonsurgical and surgical treatment of periodontitis:how many options for one disease?[J].Periodontol 2000,2017,75(1):152.
• [11] 成思源.中西医结合治疗牙周炎的研究进展[J].医疗装备，2021,34(14):187.
• [12] 杜瑞.中药提取物及方剂治疗牙周炎的研究进展[J].光明中医，2020,35(24):4012.
• [13] 国家中医药管理局.关于发布《古代经典名方目录(第一批)》的通知[EB/OL].[2021-09-01].http://kjs.satcm.gov.cn/zhengcewenjian/2018-04-16/7107.html.
• [14] 崔有晨.清胃散治疗胃火热盛型牙周炎临床观察[J].中国中医药现代远程教育，2020,18(8):97.
• [15] 李兵，侯酉娟，刘思鸿，等.经典名方复方制剂研发的文献考证要点与策略[J].中国实验方剂学杂志，2019,25(21):1.
• [16] XU X,ZHANG W,HUANG C,et al.A novel chemometric method for the prediction of human oral bioavailability[J].Int J Mol Sci,2012,13(6):6964.
• [17] 杨珏，罗霄，刘芳，等.基于网络药理学和分子对接分析厚朴治疗消化性溃疡的作用机制[J].中国中药杂志，2021,46(17):4522.
• [18] 曾玉，韩瑞婷，周庆伟.基于网络药理学与分子对接技术探讨痰热清注射液治疗急性肺损伤的作用机制[J].中国中药杂志，2021,46(15):3960.
• [19] 孟颖，徐泳，严国俊，等.清胃散加减方治疗牙周炎的Meta分析[J].世界中医药，2021,16(10):1539.
• [20] 蔡懿，吴钱生，龙文玲，等.火郁发之论治放化疗后口腔炎临床观察[J].光明中医，2018,33(8):1130.
• [21] ZHAO J,FANG Z,ZHA Z,et al.Quercetin inhibits cell viability,migration and invasion by regulating miR-16/HOXA10 axis in oral cancer [J].Eur J Pharmacol,2019,847:11.
• [22] CHENG W C,HUANG R Y,CHANG C Y,et al.Ameliorative effect of quercetin on the destruction caused by experimental perio-dontitis in rats [J].J Periodontal Res,2010,45(6):788.
• [23] PATRA J K,KIM E S,OH K,et al.Antibacterial effect of crude extract and metabolites of Phytolacca americana on pathogens responsible for periodontal inflammatory diseases and dental caries [J].BMC Complement Altern Med,2014,14:343.
• [24] 郭晓雨.槲皮素对糖尿病牙周炎大鼠牙周组织的作用及其血清AGEs水平的影响[D].石家庄：河北医科大学，2020.
• [25] ZHANG R,YANG J,WU J,et al.Berberine promotes osteoge-nic differentiation of mesenchymal stem cells with therapeutic potential in periodontal regeneration [J].Eur J Pharmacol,2019,851:144.
• [26] 张小恒.盐酸小檗碱对人牙周膜细胞生物学活性及大鼠牙周组织IL-1β、TNF-α表达的影响[D].兰州：兰州大学，2007.
• [27] MARTIN B J,CAMPBELL P M,TERRY T D,et al.A rando-mized controlled trial evaluating antioxidant-essential oil gel as a treatment for gingivitis in orthodontic patients [J].Angle Or-thod,2016,86(3):407.
• [28] 赵颖，邓云贞，朱春晖，等.阿魏酸钠改善伴糖尿病牙周炎大鼠牙周组织微循环[J].口腔生物医学，2018,9(4):173.
• [29] 陈育华，周克元，袁汉尧.山奈酚药效的研究进展[J].广东医学，2010,31(8):1064.
• [30] BALLI U,CETINKAYA B O,KELES G C,et al.Assessment of MMP-1,MMP-8 and TIMP-2 in experimental periodontitis treated with kaempferol [J].J Periodontal Implant Sci,2016,46(2):84.
• [31] ZHANG Z,SHUAI Y,ZHOU F,et al.PDLSCs regulate angiogenesis of periodontal ligaments via VEGF transferred by exosomes in periodontitis [J].Int J Med Sci,2020,17(5):558.
• [32] 胡运苑，苏黎，梁惠均.VEGF和ICAM-1在慢性牙周炎患者牙龈组织中的表达及意义[J].现代诊断与治疗，2019,30(3):347.
• [33] FANG L,CHENG J C,CHANG H M,et al.EGF-like growth factors induce COX-2-derived PGE2 production through ERK1/2 in human granulosa cells[J].J Clin Endocrinol Metab,2013,98(12):4932.
• [34] 徐颖，孟键.慢性牙周炎患者表皮生长因子表达的免疫组织化学研究[J].山西医药杂志，2016,45(12):1395.
• [35] 姜梅，徐俊仙，翁庭静.血清脂联素、超敏C-反应蛋白、白细胞介素6、肿瘤坏死因子-α联合检测在慢性牙周炎患者早期诊断中的应用价值[J].中国卫生检验杂志，2020,30(18):2248.
• [36] 黄静文.慢性牙周炎患者外周血T淋巴细胞中PD-1和PD-1L的表达量及其与炎症反应程度的相关性[J].海南医学院学报，2017,23(2):262.
• [37] SHOKOHI T,ASLANI N,AHANGARKANI F,et al.Candida infanticola and Candida spencermartinsiae yeasts:possible emerging species in cancer patients[J].Microb Pathog,2018,115(21):353.
• [38] SUI L,WANG J,XIAO Z,et al.ROS-scavenging nanomaterials to treat periodontitis [J].Front Chem,2020,8:595530.
• [39] 余海波，袁晓，郭庆圆.转录因子HIF-1α与口腔疾病关系的研究进展[J].临床口腔医学杂志，2018,34(2):125.
• [40] BERINGER A,NOACK M,MIOSSEC P.IL-17 in chronic inflammation:from discovery to targeting [J].Trends Mol Med,2016,22(3):230.
• [41] DUTZAN N,ABUSLEME L.T helper 17 cells as pathogenic drivers of periodontitis [J].Adv Exp Med Biol,2019,1197(1):107.
• [42] BUNTE K,BEIKLER T.Th17 Cells and the IL-23/IL-17 axis in the pathogenesis of periodontitis and immune-mediated inflammatory diseases [J].Int J Mol Sci,2019,20(14):3394.