敦煌医方大补脾汤对胃癌的体外活性及其配伍规律的计算机辅助药物设计分析

Analysis of in vitro activity and compatibility of Dunhuang Yifang Dabupi Decoction on gastric cancer based on computer-aided drug design

基于计算机辅助药物设计结合复杂网络分析的研究体系,分析敦煌大补脾汤防治胃癌的潜在作用机制,并通过功效分组探讨其组方规律的科学内涵。通过细胞实验研究大补脾汤冻干粉溶液对胃癌细胞增殖活性的影响;利用TCMSP、TCMID数据库收集大补脾汤化合物成分,Swiss Target Prediction预测化合物潜在作用靶标,DrugBank、GeneCards、TTD、DisGeNET收集胃癌的潜在作用靶标,通过STRING数据库分析潜在作用靶点的蛋白互作。DAVID数据库进行KEGG富集分析;根据功效特点将大补脾汤分为温中组(干姜)、益气组(人参、甘草、白术)、养阴组(麦冬、五味子)和降逆组(旋覆花)4个功效配伍组,利用Cytoscape分别构建“药味-潜在活性成分-关键靶点”网络,分析功效分组配伍规律的科学内涵;采用Schr?dinger软件进行核心成分和核心靶点的分子对接验证,并通过结合模式分析和结合自由能计算挖掘大补脾汤发挥防治胃癌作用的物质基础,通过分子动力学模拟从动力学角度分析核心成药性靶点与代表潜在化合物的相互结合模式。细胞实验证实大补脾汤冻干粉溶液能够下调AGS胃癌细胞线粒体膜电位,使细胞周期被阻滞在G_(0)/G_(1)期(P<0.05),抑制其增殖(P<0.05)。大补脾汤所含4个功效组富集的通路主要分布于机体能量代谢、炎症-免疫系统调节和周期-凋亡功能3个模块,各模块间通过共有靶标基因联结又各有侧重。分子对接结果显示,化合物甘草素、槲皮素、山柰酚、异鼠李素、麦冬高异黄酮A等可能是大补脾汤起效的多靶点成分;雌激素受体1、雄激素受体、ATP结合盒转运蛋白亚家族G2、表皮生长因子受体、糖原合酶激酶3β等靶点与各个潜在活性化合物之间的亲和力较好,可能是大补脾汤防治胃癌的潜在作用靶点,其中山柰酚与成药性靶点EGFR有较好结合能力,具有较好的结合稳定性。本研究基于计算机辅助药物设计结合复杂网络分析策略初步揭示了大补脾汤辅助防治胃癌的物质基础和分子机制,并探讨了大补脾汤不同功效组协同防治胃癌的科学内涵,为其临床应用提供化学生物信息学依据。

Based on the research system of computer-aided drug design combined with complex network analysis,the potential mechanism of Dunhuang Dabupi Decoction in preventing and treating gastric cancer(GC)is analyzed,and the scientific connotation of its prescription rules is explored through the efficacy grouping.To study the effect of Dabupi Decoction freeze-dried powder solution on the proliferation activity of gastric cancer cells through cell experiments;the TCMSP and TCMID databases were used to collect the compound components of Dabupi Decoction.Swiss Target Prediction is used to predict potential targets of compounds.DrugBank,GeneCards,TTD,and DisGeNET were used to collect potential targets for gastric cancer.Analyze protein interactions of potential targets through the STRING database.DAVID database was used for KEGG enrichment analysis;Dabupi Decoction was divided into Wenzhong group(dried ginger),Yiqi group(ginseng,licorice,Atractylodes macrocephala),nourishing Yin group(Ophiopogon japonicus,Schisandra)and Jiangni group according to its efficacy characteristics.The inverse group(inula)has 4 functional compatibility groups.Cytoscape was used to construct a network of"medicinal flavor-potential active ingredient-key target"respectively,and the network was used to analyze the scientific connotation of the compatibility of efficacy groups.The Schr?dinger software was used to verify the molecular docking of the core components and the core targets.The material basis of the Dabupi Decoction to prevent and treat gastric cancer was discovered through the combination of pattern analysis and combined free energy calculation.The core drug was analyzed from the perspective of dynamics through molecular dynamics simulation.Potential targets and representative potential compounds interact with each other.Cell experiments confirmed that Dabupitang freeze-dried powder solution can down-regulate the mitochondrial membrane potential of AGS gastric cancer cells,block the cell cycle in the G_(0)/G_(1) phase(P<0.05),and inhibit its proliferation(P<0.05).The pathways enriched by the four functional groups contained in Dabupi Decoction are mainly distributed in the body’s energy metabolism,inflammation-immune system regulation,and cycle-apoptotic functions.Each module is connected by a common target gene and has its own focus.The results of molecular docking showed that the compounds liquiritigenin,quercetin,kaempferol,isorhamnetin,methylophiopogonanone A,etc.may be the effective multi-target components of Dabupi Decoction.Estrogen receptor 1,androgen receptor,ATP-binding cassette superfamily G member 2,epidermal growth factor receptor,glycogen synthase kinase-3 beta and other targets have good affinity with each potential active compound,which may be a potential target of Dabupi Decoction for preventing and treating gastric cancer.Among them,kaempferol and the drug target EGFR not only have good binding ability,but also have good binding stability.This study is based on computer-aided drug design combined with complex network analysis strategies to initially reveal the material basis and molecular mechanism of Dabupi Decoction in the prevention and treatment of gastric cancer.It also explores the scientific connotation of Dabupi Decoction in the prevention and treatment of gastric cancer with different efficacy groups,and its clinical application provide chemical bioinformatics basis.