Identifying the molecular basis of Jinhong tablets against chronic superficial gastritis via chemical profile identification and symptom-guided network pharmacology analysis

Chronic superficial gastritis(CSG)is a common disease of the digestive system that possesses a serious pathogenesis.Jinhong tablet(JHT),a traditional Chinese medicine(TCM)prescription,exerts therapeutic effects against CSG.However,the molecular basis of its therapeutic effect has not been clarified.Herein,we employed ultra-performance liquid chromatography coupled with quadrupole time-of-flight tandem mass spectrometry(UPLC-Q/TOF-MS)based chemical profile identification to determine the chemical components in JHT.Further,we applied network pharmacology to illustrate its molecular mechanisms.A total of 96 chemical constituents were identified in JHT,31 of which were confirmed using reference standards.Based on the bioinformatics analysis using the symptom-guided pharmacological networks of“chi,”“blood,”“pain,”and“inflammation,”and target screening through the interaction probabilities between compounds and targets,matrix metalloproteinase 2(MMP2),dopamine d2 receptor(DRD2),and Aldo-keto reductase family 1 member B1(AKR1B1)were identified as key targets in the therapeutic effect exhibited by JHT against CSG.Moreover,according to the inhibitory activities presented in the literature and binding mode analysis,the structural types of alkaloids,flavonoids,organic acids,including chlorogenic acid(10),caffeic acid(13),(-)-corydalmine(33),(-)-isocorypalmine(36),isochlorogenic acid C(38),isochlorogenic acid A(41),quercetin-3-O-a-L-rhamnoside(42),isochlorogenic acid B(47),quercetin(63),and kaempferol(70)tended to show remarkable activities against CSG.Owing to the above findings,we systematically identified the chemical components of JHT and revealed its molecular mechanisms based on the symptoms associated with CSG.

Integration of network pharmacology with experimental verification reveals the hypoglycemic mechanism of coptisine in Jinqi Jiangtang tablets:inhibition of the FoxO1 signaling pathway and hepatic gluconeogenesis

Background:Jinqi Jiangtang tablets(JQJT)have been approved for the treatment of type 2 diabetes mellitus(T2DM)in China for many years.Exploring the effective substances and mechanisms of JQJT is important for its clinical application and further drug research and development.This study aimed to explore the chemical basis and mechanisms of JQJT in the treatment of T2DM.Methods:With network pharmacology,we screened substances in JQJT and their possible targets,then constructed the action network and enriched the biological functions and pathways associated with the active components,and identified the potential targets and mechanisms of JQJT in the treatment of T2DM.Based on the network pharmacology data,we explored the hypoglycemic mechanisms of coptisine in JQJT through western blot and quantitative real-time polymerase chain reaction.Results:Forty-three compounds with good pharmacokinetic properties were identified in JQJT,together with 146 potential biological targets.Among these potential targets,74 were associated with treatment of T2DM.A compound-target network of the 43 compounds against T2DM was constructed.Biological process and signal pathway enrichment analysis of the network highlighted the FoxO signaling pathway.Western blot and quantitative real-time polymerase chain reaction results showed that coptisine,but not epiberberine,significantly inhibited expression of key genes involved in hepatocyte gluconeogenesis by regulating the FoxO1 signaling pathway.Conclusion:Network pharmacology analysis and cell experiments showed that coptisine regulated glucose homeostasis by inhibiting the FoxO1 signaling pathway and hepatic gluconeogenesis,which may be one of the mechanisms of JQJT in the treatment of T2DM.

To explore the mechanism of Naodesheng tablets in the treatment of atherosclerosis based on network pharmacology and bioinformatics

Background:Naodesheng tablets(NDST)was widely used in clinical treatment as a drug for cardiovascular diseases,but the mechanism for treating atherosclerosis was unknown.On the basis of network pharmacology and bioinformatics,predict the relevant targets and signalling pathways for NDST in the treatment of atherosclerosis.Methods:First,the targets of NDST and the targets for treating atherosclerosis were looked for in different databases.Next,Venny 2.1.0 was used to find the genes that overlapped between NDST and targets for treating atherosclerosis.Subsequently,the herb-active ingredient-target-disease were obtained to explore the hub compound.Furthermore,the Metascape database was used for Gene Ontology and Kyoto Encyclopedia of Genes and Genomes enrichment analysis.And we constructed the“active ingredient-intersection target-pathway”network by Cytoscape software to gain the hub genes and pathways.Finally,molecular docking was used to verify the affinity of hub ingredients and hub genes.Results:In the results,67 active ingredients and 322 targets of NDST were selected in ingredients-targets network.154 overlapping targets of NDST(322)and atherosclerosis(1330)were obtained.Then,the herb-active ingredient-target-disease showed that quercetin,apigenin and luteolin were the hub ingredients to treat atherosclerosis.Further,the hub genes(PTGS2,RXRA,CASP3)and pathways(lipid and atherosclerosis)were accessed in active ingredient-intersection target-pathway.Finally,the results indicated that quercetin,apigenin and luteolin were better binding the PTGS2,RXRA,CASP3,especially PTGS2 and luteolin in molecular docking.Conclusion:In conclusion,quercetin,apigenin and luteolin,as the main ingredients of NDST could play a important role in PTGS2,RXRA,and CASP3 for treating atherosclerosis via the lipid and atherosclerosis(TNF signaling pathway).

基于网络药理学和分子对接探讨雷公藤片治疗类风湿关节炎的作用机制

目的:基于网络药理学和分子对接,探讨雷公藤片治疗类风湿关节炎的作用机制。方法:基于中药系统药理学数据库与分析平台,检索雷公藤片药材的化学成分和作用靶点;收集GeneCards、DisGeNET数据库中类风湿关节炎的相关靶点;筛选雷公藤片和类风湿关节炎的交集靶点,并构建蛋白质-蛋白质相互作用(PPI)网络;在Cytoscape软件中筛选PPI网络的核心靶点,对核心靶点进行基因本体(GO)功能富集分析和京都基因与基因组百科全书(KEGG)通路富集分析。结果:筛选得到雷公藤片的30个活性成分和148个靶点,与GeneCards、DisGeNET数据库中类风湿关节炎靶点的交集靶点共有77个。通过Cytoscape软件筛选出23个核心靶点,包括肿瘤坏死因子(TNF)、前列腺素内过氧化物合成酶2、胱天蛋白酶3、基质金属蛋白酶9、立早基因、人体表皮生长因子受体、丝裂原激活的蛋白激酶(MAPK)14和血管内皮生长因子A等,雷公藤甲素、豆甾醇、雷公藤二萜酸、山柰酚和β-谷固醇等为核心化合物。GO功能富集分析得到840条结果,其中生物过程739条,细胞组成29条,分子功能72条。KEGG通路富集得到相关通路133条,结果显示,雷公藤片调节卡波西肉瘤相关疱疹病毒感染、TNF信号通路、破骨细胞分化、白细胞介素17信号通路及MAPK信号通路等多条通路。结论:通过网络药理学探讨了雷公藤片多成分、多靶点、整体调节的作用特点,预测了雷公藤片可能通过抗炎、免疫调节等方面治疗类风湿关节炎。本研究为雷公藤片活性成分的生物学机制研究提供了理论依据。

Efficacy of Danlou tablet(丹蒌片)on myocardial ischemia/reperfusion injury assessed by network pharmacology and experimental verification

OBJECTIVE:To investigate the potential pharmacological mechanism of Danlou tablet(丹蒌片,DLT)with a long-term clinical application in the treatment of myocardial ischemia/reperfusion(I/R)injury through network pharmacology,molecular docking and experimental verification.METHODS:The main chemical ingredients in DLT were retrieved from the Traditional Chinese Medicine(TCM)System Pharmacology Database,the TCM information database,the bioinformatics analysis tool for molecular mechanism of TCM,and HERB database.Disease targets of I/R were accessed from the databases of Online Mendelian Inheritance in Man,Gene Cards,Therapeutic Target Database,and Dis Ge NET database.The overlaying genes of DLT and I/R were obtained from the Venny online platform.The core targets and proteinprotein interaction network were constructed and analyzed via the Search Tool for the Retrieval of Interacting Genes Proteins database and Cytoscape software.Furthermore,Gene Ontology(GO)analysis and Kyoto Encyclopedia of Genes and Genomes(KEGG)pathway enrichment analysis were performed by the Metascape platform.Based on the results,the component-target-pathway network was constructed and drafted via the Cytoscape software and the platform of Bioinformatics.Furthermore,we performed molecular docking to predict the binding information between chemical molecules and target proteins.Finally,oxygenglucose deprivation/recovery(OGD/R)-induced H9c2 cardiomyocytes were used to validate the results of network pharmacology in vitro.RESULTS:A total of 189 active chemical components in DLT and 849 correlative targets of I/R were screened.Of note,133 overlaying genes found from the Venny online platform were concentrated into 28 core genes.Furthermore,the GO and KEGG pathway enrichment analysis presented that DLT might participate in 42 types of GO molecular functions,747 types of GO biological processes,19 types of GO cellular components,and 140 kinds of pathways to treat I/R.In the component-targetpathway network,the indirect relationship between herbs and their possible effective pathways was clarified.Based on the molecular docking,we speculated that Baicalein-prostaglandin G/H synthase 2(PTGS2)with-3.24 kcal/mol,Luteolin-heat shock protein 90 alpha family class A member 1(HSP90AA1)with-3.22 kcal/mol,Baicalein-HSP90AA1 with-3.13 kcal/mol,and QuercetinHSP90AA1 with-3.05 kcal/mol possessed the strongest binding force of less than-3 kcal/mol,sequentially.Experimental verification showed that Quercetin,Luteolin,and Baicalein could increase the relative cell viability of OGD/R-stimulated cardiomyocytes,probably by suppressing PTGS2,and activating HSP90AA1 and estrogen receptor 1 expression.CONCLUSIONS:We predicted the potential active compounds as the material basis of DLT that may provide a new approach to elucidate the novel pharmacological mechanism underlying the treatment of cardiac I/R damage.

基于网络药理学及细胞实验验证探讨补肾益心片治疗糖尿病肾病的作用机制

【目的】探讨补肾益心片治疗糖尿病肾病的作用机制。【方法】应用网络药理学预测补肾益心片治疗糖尿病肾病的关键靶点。构建高糖处理MPC-5细胞模型,应用酶联免疫吸附法(ELISA)、实时定量聚合酶链反应(RT-qPCR)法和Western Blot法验证补肾益心片治疗糖尿病肾病的潜在作用靶点及通路。【结果】获得补肾益心片24个有效成分。补肾益心片治疗糖尿病肾病涉及靶点131个,通路富集分析结果显示关键靶点可能主要集中在炎症通路、脂质与动脉粥样硬化等。进一步实验验证结果发现:与高糖组比较,补肾益心片含药血清中、高剂量组MPC-5细胞上清中基质金属蛋白酶1(MMP-1)分泌水平升高(P<0.001),组织金属蛋白酶抑制因子1(TIMP-1)和转化生长因子β1(TGF-β1)分泌水平降低(P<0.001);补肾益心片含药血清高剂量组MMP-1和BCL-2蛋白及mRNA表达水平升高(P<0.001),磷脂酰肌醇3-激酶(PI3K)和蛋白激酶B(AKT)蛋白及mRNA表达水平降低(P<0.001)。【结论】补肾益心片可能是通过调节PI3K/AKT/BCL-2通路抑制肾小球系膜细胞增殖,促进细胞外基质的降解,从而发挥对糖尿病肾病的治疗作用。

基于HPLC-Q-TOF MS/MS和网络药理学的参蓉蛹草片质量标志物研究

目的:应用高效液相色谱-四极杆飞行时间质谱法(HPLC-Q-TOF-MS/MS)建立参蓉蛹草片化学成分的检测方法,并结合网络药理学和分子对接方法分析其抗疲劳的网络调控机制、预测参蓉蛹草片的质量标志物。方法:运用HPLC-Q-TOF-MS/MS对参蓉蛹草片物质基础进行分析鉴定。通过PubChem获取所选化合物的结构,利用TCMSP数据库及SwissTargetPrediction获取作用靶点;通过GeneCards、TTD、OMIM数据库筛选与疲劳相关的疾病靶点,获得药物和疲劳的交集靶点;运用UniProt数据库校准靶点名称;利用STRING数据库构建交集基因蛋白质-蛋白质相互作用(PPI)网络,并运用Cytoscape 3.8. 0软件对PPI网络进行拓扑分析;最后借助Metascape在线分析核心交集基因,进行基因本体(GO)功能富集分析、京都基因与基因组百科全书(KEGG)通路富集分析,并对分析结果进行可视化。结果:通过HPLC-Q-TOF MS/MS共得到参蓉蛹草片124个化学成分,利用网络药理学对筛选出的23个化学成分进行生物信息学分析,发现其可作用于195个靶点干预33条信号通路。分子对接结果显示,活性化合物可进入受体靶点活性空腔,与其形成的对接构象合理。结论:参蓉蛹草片可通过多成分、多靶点、多途径发挥抗疲劳作用,基于HPLC-Q-TOF MS/MS,结合网络药理学和分子对接方法初步将人参皂苷类、苯乙醇苷类预测为参蓉蛹草片的质量标志物。

基于网络药理学研究孜亚比提片对2型糖尿病的潜在分子机制

目的 利用网络药理学方法研究孜亚比提片治疗2型糖尿病(T2DM)的分子机制。方法 从TCMSP、ETCM、CNKI中得到孜亚比提片化学成分和药物靶点,利用OMIM和GeneCards数据库筛选T2DM的疾病靶点;使用Cytoscape 3.6.1软件构建“药物-成分-靶点网络”筛选出核心成分;运用蛋白质相互作用网络筛选出核心靶点;通过DAVID数据库进行GO和KEGG富集分析。结果研究共收集到孜亚比提片活性成分144个,度值排名靠前的为槲皮素、山柰酚、异鼠李素等。活性成分相关靶点823个,其中与T2DM相关的有700个,包含为SRC、MAPK1、MAPK3等。GO功能分析提示与信号传导、蛋白质磷酸化以及蛋白质结合等分子功能有关。KEGG通路富集分析所涉及的主要信号通路有脂质与动脉粥样硬化、糖尿病并发症中的AGE-RAGE信号通路及HIF-1信号通路等。结论 孜亚比提片治疗T2DM具有多成分、多靶点、多通路协同干预的特点,其主要由槲皮素、山柰酚、异鼠李素等成分通过作用SRC、MAPK1、MAPK3靶点调控AGE-RAGE信号通路发挥综合治疗作用。

基于网络药理学方法探讨杞蓉片治疗早发性卵巢功能不全的作用机制

目的:采用网络药理学方法探讨杞蓉片治疗早发性卵巢功能不全(premature ovarian insufficiency,POI)的作用机制。方法:在中药药理学数据库与分析平台(traditional Chinese medicine systems pharmacology database and analysis platform,TCMSP)中获取杞蓉片的有效成分及相应靶点,同时查询人类基因数据库(Genecards)等数据库,获取疾病相关的靶点,并利用韦恩图(Venn)图与药物作用靶点取交集,筛选杞蓉片治疗POI的关键靶点。利用STRING平台建立预测靶蛋白之间的相互关系网络,根据联系密切程度筛选出核心靶标,用R语言进行核心靶标的基因本体论功能富集分析(gene ontology,GO)和京都基因与基因组百科全书富集分析(kyoto encyclopedia of genes and genomes,KEGG)。结果:收集到符合筛选条件的杞蓉片活性成分102个,染料木素、槲皮素、芹菜素、β-谷固醇等化合物可能为其作用的主要活性成分,药物与POI共同靶点243个,其中肿瘤蛋白p53基因、丝氨酸/苏氨酸激酶1、转录激活因子3、肿瘤坏死因子、表皮生长因子受体、白细胞介素6在促分裂素原活化蛋白激酶、血管内皮生长因子为核心靶基因,通过KEGG通路分析显示,杞蓉片治疗POI的生物信号通路主要集中在磷脂酰肌醇3激酶和蛋白激酶B、缺氧诱导因子1等。分子对接结果显示大部分靶点与成分结合活性较强。结论:杞蓉片通过多种成分、多核心靶点参与了一系列通路,通过发挥多种生物、分子过程来治疗POI。

基于网络药理学和分子对接的养心氏片治疗冠心病合并抑郁症机制

通过网络药理学和分子对接技术探讨养心氏片(Yangxinshi,YXS)治疗冠心病(coronary heart disease,CHD)合并抑郁症(major depressive disorder,MDD)的机制。使用TCMSP和TCM-ID数据库筛选YXS中的活性化合物及其相关靶点,并在Uniport数据库中获取靶点的标准基因名;通过使用GeneCards和OMIM数据库,收集与冠心病(CHD)和抑郁症(MDD)相关的疾病靶点,对药物靶点和疾病靶点进行交集分析,以获得蛋白质互作关系;利用David数据库进行基因本体(GO)和京都基因与基因百科全书信号通路富集分析(KEGG);最后使用AutodockTools软件进行分子对接验证。富集分析显示YXS中的295个活性成分(如槲皮素、木犀草素、山柰酚和豆甾醇)作用于STAT3、IL1B、IL6等290个靶点。此外,YXS还涉及AGE-RAGE信号通路、脂质与动脉粥样硬化信号通路、流体剪切应力与动脉粥样硬化信号通路等。分子对接结果显示良好的对接效果。YXS治疗CHD合并MDD的药理作用主要通过作用于槲皮素、木犀草素、山柰酚和豆甾醇等活性成分的STAT3、IL1B、IL6等靶点,通过AGE-RAGE、脂质与动脉粥样硬化等信号通路抑制炎症反应,提高5-羟色胺(5-HT)含量等。研究结果为后续的实验研究和临床应用提供了理论基础。

基于网络药理学、分子对接及实验验证探讨滋肾健脾化瘀片对糖尿病视网膜病变的干预机制

目的运用网络药理学方法和分子对接技术探讨滋肾健脾化瘀片(山萸肉、三七、黄芪、葛根、鸡血藤、生地黄)治疗糖尿病视网膜病变(DR)的作用机制,并通过体外实验进行验证。方法(1)利用中药系统药理学数据库与分析平台(TCMSP)及BATMAN-TCM数据库筛选滋肾健脾化瘀片的有效成分及其对应的靶点蛋白。利用GeneCards、OMIM及TTD数据库检索DR疾病相关靶点。利用VENNY 2.1.0平台对药物活性成分靶点与DR疾病相关靶点取交集(共同靶点),即为滋肾健脾化瘀片治疗DR的潜在作用靶点。构建“中药-活性成分-共同靶点”网络,筛选出滋肾健脾化瘀片治疗DR的关键活性成分。将共同靶点导入STRING数据库,获取PPI网络关系,并筛选出核心靶点。运用Metascape平台对共同靶点进行GO功能及KEGG通路富集分析。将关键活性成分及核心靶点通过Autodock 4软件进行分子对接验证。(2)制备滋肾健脾化瘀片含药血清及空白血清。将人视网膜微血管内皮细胞(HRmECs)随机分成5组:对照组(低糖DMEM培养基+10%空白血清)、高糖组(高糖DMEM培养基+10%空白血清)及滋肾健脾化瘀片含药血清低、中、高剂量组(高糖DMEM培养基+10%低、中、高剂量含药血清),培养48 h后进行检测。采用CCK-8法检测HRmECs细胞增殖活性;RT-qPCR法检测HRmECs细胞中IL-1β、AKT1、VEGFA、TP53 mRNA表达水平。结果(1)共筛选出滋肾健脾化瘀片治疗DR的潜在作用靶点(共同靶点)74个;9个关键活性成分包括槲皮素、芒柄花黄素、毛地黄黄酮、β谷甾醇、山柰酚、毛蕊异黄酮、γ-氨基丁酸、豆甾醇、异鼠李亭;12个核心靶点:IL-1β、PPARG、NOS3、CXCL8、IL-6、AKT1、TNF、INS、EGF、VEGFA、TP53、PTGS2。GO功能及KEGG富集分析显示核心靶点主要参与了炎症反应、蛋白质磷酸化的正调控、细胞迁移等生物过程,以及NF-κB信号通路、AGE-RAGE信号通路、HIF-1通路、TNF通路、PI3K-AKT通路等。关键活性成分与核心靶点均具有较好的结合亲和力。(2)与对照组比较,高糖组HRmECs细胞活性显著降低(P<0.01);细胞中VEGFA、TP53、IL-1βmRNA表达水平显著升高(P<0.01),AKT1 mRNA表达水平显著降低(P<0.01)。与高糖组比较,滋肾健脾化瘀片含药血清低、中、高剂量组的HRmECs细胞活性均显著升高(P<0.05,P<0.01),并呈浓度依赖性;含药血清高剂量组细胞的VEGFA、TP53、IL-1βmRNA表达水平显著降低(P<0.05,P<0.01),而AKT1 mRNA表达水平显著升高(P<0.01)。结论滋肾健脾化瘀片可能通过槲皮素、山柰酚、毛地黄黄酮等多种活性成分,作用于IL-1β、IL-6、VEGFA等核心靶点,以及NF-κB信号通路、AGE-RAGE信号通路、PI3K-AKT通路等关键通路,从而发挥对DR的治疗作用。

基于网络药理学和分子对接技术探究定清片活性成分治疗白血病的作用机制

目的探讨定清片治疗白血病的药理作用机制。方法检索中药系统药理学数据库与分析平台(TCMSP)、本草组鉴平台(HERB),收集定清片的活性成分;利用SwissTargetPrediction数据库预测其作用靶点。利用GeneCards、OMIM和DisGeNET数据库获取疾病靶点,将药物与疾病的共有靶点导入String网站,构建共有靶点蛋白相互作用(PPI)网络,并利用MCODE插件筛选核心子网络。利用DAVID数据库中对评分最高的基因簇1基因进行GO和KEGG富集分析,构建“成分-靶点-通路”网络;利用AutoDock对定清片的活性成分和核心靶点进行分子对接,并对其结果进行可视化分析。结果数据库检索共获得定清片活性成分82个及其作用靶点439个,白血病相关靶点1878个,活性成分和疾病的共有靶点169个。由度值推测定清片中发挥作用的主要活性成分有槲皮素、木犀草素、山柰酚等,而PPI核心网络表明其治疗白血病的关键靶点主要包括TP53、MMP9、TNF、AKT1、CASP3等;子网络的基因富集分析及“成分-靶点-通路”网络图显示定清片可能通过调节TNF、IL-17等信号通路来发挥对白血病的治疗作用。分子对接结果显示活性成分与靶点间有较强的结合活性。结论定清片的活性成分可能通过调控TP53、AKT1和CASP3等基因参与TNF、IL-17等信号通路,从而发挥对白血病的治疗作用。

基于网络药理学和分子对接探讨壮骨强筋片治疗老年性骨质疏松症的作用机制

目的 运用网络药理学结合分子对接技术探讨壮骨强筋片治疗老年性骨质疏松症(SOP)的物质基础及作用机制。方法 通过TCMSP、HERB、PubChem、SwissTargetPrediction和UniProt数据库筛选壮骨强筋片组方药物的活性成分及作用靶点;检索DisGeNet、GeneCards、OMIM数据库获得SOP相关靶点;采用Cytoscape3.8.0构建“疾病-药物-成分-交集靶点”网络;利用STRING和Cytoscape3.8.0构建蛋白质-蛋白质相互作用网络;利用R4.2.0进行GO功能及KEGG通路富集分析;选取度值较高的核心成分与核心靶点进行分子对接。结果 获得壮骨强筋片作用靶点270个,SOP相关靶点433个,药物与SOP交集靶点61个;核心成分主要有槲皮素、木犀草素和山柰酚等,核心靶点有TNF、IL6、AKT1、VEGFA等。GO生物过程主要涉及氧化应激反应、生殖结构发育、生殖系统发育等,KEGG通路主要富集于糖尿病并发症相关的AGE-RAGE信号通路、脂质和动脉粥样硬化、流体剪切应力和动脉粥样硬化等。结论 壮骨强筋片可能通过作用于TNF、IL6、AKT1、VEGFA等靶点,调节免疫与细胞代谢,发挥治疗SOP的作用。

基于网络药理学与分子对接技术探讨脉管复康片治疗动脉粥样硬化的作用机制

目的:利用网络药理学方法及分子对接技术探析脉管复康片治疗动脉粥样硬化(atherosclerosis,AS)的机制。方法:通过传统中药系统药理学数据库与分析平台(traditional Chinese medicine systems pharmacology database and analysis platform,TCMSP)筛选脉管复康片(丹参、鸡血藤、没药、乳香、郁金)活性成分和靶基因。使用人类基因数据库(the human gene database,GeneCards)、在线人类孟德尔遗传数据库(online mendelian inheritance in man,OMIM)、药物靶标数据库(therapeutic target database,TTD)、遗传药理学与药物基因组学数据库(pharmacogenetics and pharmacogenomics knowledge base,PharmGKB)、DrugBank疾病数据库获得AS相关基因。使用Cytoscape 3.8.2构建“成分-靶点”网络。使用STRING数据库下载蛋白-蛋白互作网络(protein-protein interactions,PPI)数据,导入Cytoscape,使用CytoNCA插件获取核心靶基因。使用R软件进行基因本体(gene ontology,GO)和京都基因与基因组百科全书(kyoto encyclopedia of genes and genomes,KEGG)富集分析。对活性成分及核心基因做分子对接。结果:从脉管复康片中共筛选出157个活性成分,对应AS靶基因238个,PPI核心靶基因共20个。GO功能富集显示生物学功能主要涉及脂多糖反应、细菌来源分子反应、金属离子反应、氧化应激反应等;KEGG通路富集显示主要涉及流体剪切应力和AS、AGE-RAGE信号通路、PI3K-Akt信号通路等;分子对接结果提示槲皮素、木犀草素、柚皮素、丹参酮ⅡA、刺芒柄花素、甘草查尔酮A与4个核心蛋白JUN、MAPK1、TP53、STAT3结合稳定。结论:脉管复康片可通过槲皮素、木犀草素等活性成分作用于多种信号通路及多个基因靶点治疗AS。

基于网络药理学和斑马鱼模型的心可舒片抗心肌缺血活性成分及作用机制研究

目的基于网络药理学和斑马鱼模型研究心可舒片抗心肌缺血活性成分及作用机制。方法利用盐酸异丙肾上腺素(isoprenaline,ISO)诱导的斑马鱼心肌缺血模型和过氧化氢(H_(2)O_(2))诱导的大鼠心肌细胞(H9c2)损伤模型,评价心可舒片抗心肌缺血活性。利用TCMSP等数据库检索心可舒片的活性成分;利用PharmMapper数据库预测潜在作用靶点;利用OMIM数据库检索心肌缺血疾病靶点;分析心可舒片抗心肌缺血的潜在治疗靶点;对核心靶点进行GO和KEGG富集分析。利用斑马鱼和细胞模型对活性成分进行验证;利用qRT-PCR检测心可舒片对关键靶点表达的影响。结果心可舒片可以明显缓解ISO诱导的斑马鱼心包水肿、心动过缓、静脉窦-动脉球(SV-BA)距离增大,提高细胞存活率。心可舒片的30个潜在活性成分主要作用于ALB、AKT1、MAPK1等30个核心靶点;通过调节蛋白质磷酸化、负调控凋亡、正调控PI3K信号转导等627个GO条目,调控PI3K/Akt、FOXO、Ras等117条信号通路发挥抗心肌缺血作用。丹酚酸A、紫草酸、迷迭香酸、丹酚酸D、丹酚酸B、人参皂苷Rg2、金丝桃苷、3′-甲氧基葛根素、3′-羟基葛根素和人参皂苷Rg1能够缓解ISO引起的斑马鱼心肌缺血、提高缺氧细胞存活率。心可舒片能够上调ras、akt1等基因的表达,下调mapk1、mapk8等基因的表达。结论心可舒片中的丹酚酸A、紫草酸等活性成分通过靶向AKT1、MAPK1等靶点,调控PI3K/Akt、MAPK、Ras等信号通路发挥抗心肌缺血作用。

基于网络药理学和HPLC法建立宠物用复方石淋通片中多指标成分的质量控制方法

建立宠物用复方石淋通片多指标成分含量测定方法。利用网络药理学方法筛选复方石淋通片治疗尿石症、膀胱炎的关键活性成分,结合相关文献及《中国药典》相关要求,确定指标性成分。利用HPLC分析方法,使用CAPCELL PAK C18 MG(S-5)4.6×250 mm色谱柱,柱温30℃,进样量10μL,流速为1.0 mL·min^(-1),检测波长335 nm,以乙腈-0.1%甲酸水为流动相,梯度洗脱,建立指标性成分含量测定的方法。通过网络药理学筛选出槲皮素、咖啡酸、绿原酸等十个关键成分。结合《中国药典》及相关文献选出新绿原酸、绿原酸、隐绿原酸、咖啡酸、夏佛塔苷、异夏佛塔苷六种指标性成分。含量测定方法学结果表明:各成分线性关系良好(r≥0.999);精密度良好,RSD为0.03%~0.21%;重复性良好,RSD为0.70%~2.02%;供试品在24h内稳定性良好,RSD为0.30%~2.02%;平均加样回收率(RSD)在98.31%~104.31%(0.36%~2.28%);三批样品中六种成分的含量分别在0.990~1.434 mg·g^(-1)、1.533~2.083 mg·g^(-1)、1.311~2.023 mg·g^(-1)、0.332~0.353 mg·g^(-1)、1.594~1.768 mg·g^(-1)、0.787~1.057 mg·g^(-1)之间。该含量测定方法有较好的重复性和稳定性,可用于宠物用复方石淋通片的质量分析。

护肝片联合抗感颗粒治疗EBV肝损伤的网络药理学分析

目的:通过网络药理学联合分子对接技术探讨护肝片联合抗感颗粒治疗EB病毒(Epstein-Barr virus,EBV)肝损伤的分子机制。方法:利用TCMSP、HERB数据库和Swiss Target Prediction平台筛选和预测护肝片联合抗感颗粒的活性成分及潜在靶点。通过GeneCards数据库检索EBV肝损伤的相关基因。运用Cytoscape 3.9.1软件构建药物活性成分-靶点网络,并筛选出关键成分。对护肝片联合抗感颗粒治疗EBV肝损伤的潜在作用靶点进行GO及KEGG通路富集分析,并将其导入STRING平台,构建PPI网络,利用MCC和MNC算法筛选出核心靶点。最后,利用CB-Dock对关键成分与核心靶点进行分子对接。结果:共收集护肝片联合抗感颗粒的活性成分96个,护肝片联合抗感颗粒治疗EBV肝损伤的作用靶点57个。GO和KEGG通路富集分析结果主要与细胞因子受体结合、细胞因子活性、人巨细胞病毒感染等功能和通路相关。分子对接结果证实核心靶点STAT3、AKT1、JUN、TP53能稳定地与关键成分槲皮素、山柰酚、木犀草素、金合欢素、异鼠李素结合。结论:护肝片联合抗感颗粒主要通过槲皮素、山柰酚、木犀草素、金合欢素和异鼠李素等关键成分作用于STAT3、AKT1、JUN和TP53等核心靶点,参与细胞因子相关生物细胞过程的调节,最终起到治疗EBV肝损伤的作用。

基于网络药理学和分子对接探讨心脉安片“异病同治”心律失常和心力衰竭的作用机制

应用网络药理学方法和分子对接技术揭示心脉安片治疗心律失常和心力衰竭的作用机制。通过TCMSP、BATMAN-TCM、Swiss Target Prediction和Stitch数据库收集心脉安片化学成分及其相关靶点,并结合GeneCards、TTD、OMIM和DrugBank数据库获取心律失常和心力衰竭疾病靶点。采用Cytoscape软件获得心脉安片“异病同治”的共同靶点,并绘制中药有效成分共同靶点疾病网络图,利用STRING数据库对疾病成分靶点构建蛋白蛋白互作(PPI)分析,通过Cytoscape软件分析PPI网络的核心靶点,并运用DAVID数据库对核心靶点进行GO功能富集分析和KEGG通路富集分析。同时将核心靶点与关键活性成分进行分子对接验证。本研究从心脉安片中筛选出146个活性化合物,其“异病同治”的共同靶点178个,核心靶点11个,分别是STAT3、AKT1、IL-6、TNF、MAPK3、TP53、EGFR、HSP90AA1、VEGFA、EDN1和PPARA。关键活性成分有槲皮素、山柰酚、木犀草素、人参皂苷Rg1、人参皂苷Re、人参皂苷Rb1和毛蕊异黄酮葡萄糖苷等成分。关键生物通路包括HIF-1信号通路和TNF信号通路等。分子对接结果显示,槲皮素、山柰酚、木犀草素、人参皂苷Rg1、人参皂苷Re、人参皂苷Rb1和毛蕊异黄酮葡萄糖苷等成分与STAT3、TNF、IL-6和VEGFA等靶点稳定对接。心脉安片“异病同治”心律失常和心力衰竭多成分、多靶点、多通路的作用机制,为进一步研究心脉安片效应机制,探索其新的临床应用提供科学依据。

补益强心片治疗慢性心力衰竭的网络药理学研究

目的:确定补益强心片的有效成分和治疗慢性心力衰竭的靶点,以研究补益强心片治疗慢性心力衰竭的潜在机制。方法:通过中药系统药理分析平台(TCMSP)数据库获得补益强心片的有效成分和预测靶点。使用疾病数据库筛选慢性心力衰竭的疾病靶点,使用在线工具Draw Venn Diagram获取药物和疾病的共同靶点,将获得的共同靶点上传到STRING数据库,用于蛋白质-蛋白质相互作用(PPI)网络分析。然后,使用Cytoscape 3.9.0软件构建药物-活性成分-靶点网络图和PPI网络的可视化分析。通过DAVID数据库进行基因本体(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。最后,使用Autodock Vina进行分子对接。结果:共获得115个药物活性成分、247个药物靶点、2 685个疾病靶点和153个与药物和疾病相关的共同靶点。通过PPI网络分析确定了蛋白激酶B1(AKT1)、白细胞介素6(IL-6)、血管内皮生长因子A(VEGFA)、肿瘤蛋白P53(TP53)、白细胞介素-1β(IL-1β)、半胱氨酸蛋白酶3基因(CASP3)、表皮生长因子受体(EGFR)、JUN、基质金属蛋白酶-9(MMP-9)和前列腺素内过氧化物合成酶2(PTGS2)等核心靶点。富集分析结果表明,潜在核心药物成分通过抑制炎症反应、调节血管生成、凋亡等生物过程和晚期糖基化终末产物(AGE)-晚期糖基化终末产物受体(RAGE)信号通路、癌症中的通路、流体切应力与动脉粥样硬化、肿瘤坏死因子(TNF)信号通路、白细胞介素-17(IL-17)信号通路等发挥治疗慢性心力衰竭的药理作用。分子对接结果表明,调控网络中的关键靶点与相关活性组分具有较高的结合活性。结论:本研究使用网络药理学方法筛选出补益强心片治疗慢性心力衰竭的主要活性化合物、关键靶点和主要通路,揭示其潜在机制,为进一步研究提供理论依据和参考价值。

Network Pharmacology-Based Exploration of the Mechanism of Guanxinning Tablet for the Treatment of Stable Coronary Artery Disease

Objective:Network pharmacology was utilized to explore the mechanism of Guanxinning(GXN)tablet for the treatment of stable coronary artery disease(SCAD).Materials and Methods:First,active ingredients and therapeutic targets were predicted by databases and gene chip.Then,we constructed the compound-target(C-T)network and target-disease(T-D)network to screen hub compounds and therapeutic targets based on contribution index(CI),degree,closeness,betweenness,and coreness in the networks.Enrichment analysis was performed on hub therapeutic targets,and finally,the verification of hub ingredients and hub therapeutic targets was performed through molecular docking.Results:With"oral bioavailability≥30%,druglikeness≥0.18,and half-life≥4 h"as screening conditions,58 active ingredients were obtained.Seven hundred and seventeen compound targets and 636 SCAD targets were retrieved using databases and gene chip,and the intersection of both(139 targets)was defined as therapeutic targets.According to CI,degree,betweenness,closeness,and coreness,2 hub compounds and 13 hub therapeutic targets were chosen from the C-T network and T-D network,respectively.The Gene Ontology and Kyoto Encyclopedia of Genes and Genomes enrichment analysis indicated that GXN treated SCAD from several aspects including inflammatory reaction,oxidative stress,nutritional metabolism,blood pressure regulation,ventricular remodeling,vascular smooth muscle proliferation,angiogenesis,and platelet aggregation.Tissue enrichment analysis revealed that the therapeutic targets were enriched in multiple organs and tissues.The excellent binding force between the hub compounds and hub therapeutic targets was verified by molecular docking.Conclusions:The treatment of SCAD by GXN has the characteristics of multiple ingredients,multiple targets,and multiple approaches.Consequently,it may theoretically treat SCAD from multiple angles and levels.