Mechanism of Zixinyin oral liquid in the treatment of insomnia based on network pharmacology and molecular docking

Objective:To explore the molecular mechanism of Zixinyin oral liquid(ZOL)in the treatment of insomnia.Methods:The compounds and action targets of four herbal medicines in ZOL were collected via Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform.Genes corresponding to the targets were queried from the UniProt database.Genecards database was searched to screen the related targets of insomnia.A Gene Ontologyfunction enrichment analysis and a Kyoto Encyclopedia of Genes and Genomespathway enrichment analysis were performed by Database for Annotation,Visualization and Integrated Discovery.AutoDock software was used for molecular docking to verify the results of network analysis.Results:A total of 47 effective compounds and 187 potential targets of ZOL were screened out from the four drugs.A total of 2592 disease targets were screened out from the genecards database,and there were 1576 genes whose relevance score≥0.5.46 genes were obtained by taking the intersection of 187 potential targets of ZOL and 1,576 targets of insomnia.A total of 3,405 entries were obtained from Gene Ontologyfunctional enrichment(P<0.05).A total of 195 signaling pathways were obtained through a Kyoto Encyclopedia of Genes and Genomespathway analysis(P<0.05).Tumor necrosis factor,interleukin 17and hypoxia inducible factor-1signaling pathways were closely related to insomnia.The results of molecular docking show that all the core compounds of ZOL had a certain degree of affinity with gamma-aminobutyric acid receptor subunit alpha-1(GABRA1)and tumor necrosis factor-α(TNF-α).Paeoniflorgenone shows the highest affinity with GABRA1,and beta-sitosterol shows the highest affinity withTNF-α.ZOLmay have a therapeutic effect on insomnia through its action on targets such as GABRA1 and TNF-α,meanwhile regulating many signaling pathways.

Preliminary exploration of the mechanism of compound Qinlan oral liquid against new coronavirus pneumonia based on network pharmacology and molecular docking technology

Objective:Applying Traditional Chinese Medicine(TCM)network pharmacology and molecular docking technology to explore the mechanism of anti-coronary virus pneumonia(Corona Virus Disease 2019,COVID-19)of Compound Qinlan oral liquid.Methods:Traditional Chinese Medicines Integrated Database(TCMID),Traditional Chinese Medicine Systems Pharmacology(TCMSP),OMIM,GeneCards,String and others online databases were used for building a series of networks,and selecting the core targets and analyzing the signal pathways.Finally,Discovery Studio 2016 software was used to conduct molecular docking of the main compounds(Chinese Medicine Legal Quality Control Compound)of Compound Qinlan oral liquid with key targets ACE2,3CLpro,etc.Results:the results showed that Compound Qinlan oral liquid has specific effects in lung,heart and stomach diseases.The Compound Qinlan oral liquid compound-pneumonia target network contained 98 compounds and 184 corresponding targets,and the core targets involved INS,TP53,IL6,VEGFA,ALB and JUN.GO(GeneOntology)function enrichment analysis yielded 653 GO entries,and KEGG(KyotoEncyclopedia of Genes and Genomes)enrichment screening yielded 112 related pathways,including hypoxia inducible factor-1(HIF-1)and Toll-like receptor(TLRs)signaling pathway related to pneumonia,as well as Influenza A signaling pathway and Hepatitis B signaling pathway related to microbial infection.The results of molecular docking show that Isochlorogenic acid C,Baicalein,etc have good binding capacity with ACE2,3CLpro,AKT1 and other proteins.Conclusion:In this paper,we preliminarily explored the potential therapeutic mechanism for Compound Qinlan oral liquid to against coronavirus pneumonia(COVID-19)and predicted the active ingredients.We hope that the results will help to further study on the active ingredients and mechanism of Compound Qinlan oral liquid for anti-COVID-19.

Exploring the mechanisms of Yuanhu Zhitong oral liquid for primary dysmenorrhea through network pharmacology,molecular docking,and molecular dynamics simulation

Objective:Primary dysmenorrhea(PD)is a prevalent gynecological disorder.Yuanhu Zhitong oral liquid(YHZT)presents a promising alternative treatment for PD.However,the mechanisms underlying its efficacy remain unclear.This study aims to investigate the potential targets and mechanisms of action of YHZT in treating PD using network pharmacology,molecular docking,and molecular dynamics simulations.Methods:Potential compounds from YHZT were obtained from the Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform(TCMSP)and the Traditional Chinese Medicine Information Database(TCMID).The relevant targets of these compounds were identified using the similarity ensemble approach(SEA)and the Swiss Target Prediction database.PD-related targets were retrieved from the Genecards,DrugBank,and Disgenet databases.ClusterProfiler was utilized for Gene Ontology(GO)and Kyoto Encyclopedia of Genes and Genomes(KEGG)pathway enrichment analysis.The compound-target-pathway(CTP)network was constructed to facilitate the identification of key compounds,core targets,and signaling pathways.Finally,molecular docking and molecular dynamics simulations were performed to evaluate the interactions between the targets and compounds.Results:A total of 153 putative compounds and 129 targets of YHZT were identified.Network topology analysis revealed eight core targets and six key compounds.The effects of YHZT were mediated by genes associated with hormone and steroid metabolism,as well as pathways involved in steroid hormone biosynthesis and cytochrome P450 enzymes.Docking results showed free binding energies ranging from-6.06 to-10.85 kcal/mol,indicating strong binding affinity between the compounds and targets.Molecular dynamics simulation results further confirmed the stability of these interactions.Conclusion:This study demonstrates that YHZT treats PD by suppressing inflammatory reactions and modulating hormone and cytochrome P450 concentrations.Key compounds such as ferulic acid,(R)-canadine,(S)-canadine,canadine,and asristolone are implicated in this process.These findings offer insights into the mechanisms underlying the action of YHZT and provide a foundation for further research in this area.

Effect of Biejiajian Oral Liquid on the Expression of RAAS in Hepatic Fibrosis Rats

Objective: To study the effect of Biejiajian Oral Liquid (BOL) on the rennin angiotensin aldosterone system (RAAS) in plasma of hepatic fibrosis rats and in hepatic stellate cell (HSC) of normal rats. We explore the mechanism of BOL on inhibiting the activation of HSC and illustrate its mechanism of anti-hepatic fibrosis further. Methods: SD Rats were divided into 5 groups randomly: normal control group, model group, Enalapril group and BOL groups with different concentration (2.0 g/ml or 1.0 g/ml). Every group was administered with CCl4 and olive oil solution to induce hepatic fibrosis except normal one. Each group was treated with the respective drug for 5 weeks and then got the blood. The level of renin, angiotensin II and aldosterone in the plasma of liver fibrosis rats were detected by the radioimmunoassay. By using reverse transcription-polymerase chain reaction (RT-PCR) to measure the gene expression of the rennin, ACE, angiotensinogen, AT1R and ALD. The AT1R gene expression in normal HSC was determined by the immunohistochemical measurement. Results: BOL could effectively reduce the activity of the PRA, AngIIand ALD, which showed a significant effect on the inhibition of the AngII (P < 0.01). Meanwhile, compared with the normal control group, there was a notable inhibitory action on the PRA of HSC which was administrated by serum containing BOL (P < 0.05). And yet, drug applied group showed no difference with the model group for other factors of the RAAS. Conclusion: BOL can inhibit the expression of RAAS in the rat plasma and can inhibit the expression of the mRNA of renin in the normal HSC, which could be the mechanism of anti-hepatic fibrosis.

Elucidation of Immune Regulation Mechanism of Renshen Guben Oral Liquid by Network Pharmacology and Molecular Docking

Objective:To investigate the mechanism of Renshen Guben oral liquid(RSGB)enhancing immune function.Materials and Methods:Network pharmacology and molecular docking were used to intuitively demonstrate the mechanism of immune regulation of RSGB.Results:A total of 112 active compounds of RSGB were found,and 501 targets were predicted.Furthermore,2974 immune targets were obtained from UniProt and NCBI Gene databases,and 111 common targets of RSGB and immunity were obtained.Among them,interleukin(IL)6,tumor necrosis factor,AKT1,VEGFA,STAT3,MAPK1,SRC,EGFR,IL1B,and PTGS2 might be the key targets for RSGB to improve immunity.ClueGO and Kyoto Encyclopedia of Genes and Genomes analysis showed that the immunoregulatory mechanism of RSGB may find a relation with the B cell receptor signaling pathway and T cell receptor signaling pathway.Furthermore,this study preliminarily explored the mechanism of RSGB improving menopausal syndrome,polycystic ovary syndrome,and cancer-related fatigue by enhancing immunity.Conclusions:RSGB can improve the body's immunity through multicomponent,multitarget,and multipathway.In addition,RSGB can also improve the immune capacity of the body to assist in the treatment of diseases,which has great potential as an immunomodulator.

基于网络药理学和分子对接探讨参枝苓口服液治疗阿尔茨海默病的作用机制

目的:基于网络药理学和分子对接技术探讨参枝苓口服液治疗阿尔茨海默病(AD)的作用机制。方法:利用中药系统药理学分析平台(TCMSP)和中药分子机制的生物信息学分析工具数据库(BATMAN-TCM)查找与参枝苓口服液中中药相关的化学成分及其作用靶点;通过DisGeNET数据库获取与AD相关的靶标,并进一步筛选得到参枝苓口服液治疗AD的潜在作用靶点;利用Cytoscape软件构建成分-靶点相互作用网络,确认可能的关键靶点和关键通路;采用DAVID数据库对筛选出的潜在作用靶点进行基因本体(GO)功能分析及京都基因与基因组百科全书(KEGG)代谢通路富集分析;通过分子对接预测活性成分与靶蛋白的结合能力。结果:在参枝苓口服液中,共筛选出34个与AD相关的潜在活性成分和56个潜在作用靶点;GO生物功能分析共包含541条富集结果,主要涉及基因表达的正向调节、同源蛋白结合等;KEGG富集得到214条代谢通路,主要包括AD及自噬信号通路等。分子对接结果表示β-谷甾醇、山柰酚、木犀草素与半胱氨酸蛋白酶3(CASP3)、前列腺素内过氧化物合酶(PTGS2)、白细胞介素-6(IL-6)、肿瘤坏死因子(TNF)、B细胞淋巴瘤/白血病-2(Bcl-2)均可较好结合,可能是参枝苓口服液抗AD最有效的活性成分。结论:网络药理学及分子对接研究结果揭示了参枝苓口服液治疗AD的多成分、多靶点、多途径的作用特点,并预测了其可能的活性成分、作用通路和关键靶点,为其药效物质基础和作用机制提供了理论基础。

基于网络药理学方法探讨四磨汤口服液对腹部手术患者的胃肠功能恢复作用

目的基于网络药理学方法探讨四磨汤口服液在腹部手术后促进肠胃功能恢复的潜在作用机制。方法采用ETCM、TCMSP数据库筛选出四磨汤口服液的活性成分,利用TCMSP靶点预测模型预测活性成分的潜在靶点;利用Genecards、OMIM、Disgenet和DigSee等数据库检索手术后促进肠胃功能恢复的相关靶点;利用Biogenet分别对药物与疾病靶点进行蛋白质相互作用网络构建,并提取交集网络以获得四磨汤治疗胃肠功能紊乱的关键靶点;利用Metascape对关键靶点进行GO和KEGG富集分析。结果得到药效成分37个、药物靶点241个、疾病靶点1315个,最终获得关键靶点24个、KEGG通路144条。四磨汤口服液治疗手术后胃肠功能障碍主要成分为槲皮素、橙皮苷等,关键靶点为ALB、MAPK14、AKT1、MAPK8、EGFR、IGF1、NR3C1、RHOA,关键的生物学进程和通路可能包括PI3K-Akt信号通路、NF-κB信号通路、PPAR信号通路等。结论本研究初步揭示了四磨汤多成分、多靶点、多途径治疗腹部手术后胃肠功能紊乱的机制,为四磨汤的临床合理应用提供了依据。

基于网络药理学和分子对接探究芪芝口服液提高家禽免疫功能的作用机制

试验旨在探究芪芝口服液提高家禽免疫功能的作用机制。采用中药系统药理学数据库与分析平台(TCMSP)收集该复方中黄芪和灵芝2味药材的活性成分及其作用靶点,并与OMIM、TTD和GeneCards数据库收集免疫的靶点进行交集,获得芪芝口服液作用于家禽免疫的潜在靶点。运用Cytoscape 3.9.1软件构建中药-成分-靶点-通路-疾病网络图和蛋白质相互作用(PPI)网络,并对靶点进行GO富集和KEGG通路分析。使用分子对接技术进行活性成分和核心靶点的分子对接验证。结果显示,芪芝口服液30个成分对应66个靶点,家禽免疫329个靶点,其中交集靶点17个,PPI核心靶点包括核因子κB抑制因子α(NFKBIA)、白细胞介素-6(IL-6)和干扰素-γ(IFNG)等。GO功能富集共有38条结果,包括33个生物过程条目、4个分子功能条目和1个细胞成分条目;KEGG通路分析显示交集基因主要富集于C型凝集素受体信号通路、FoxO信号通路和Toll样受体等信号通路。分子对接结果显示,关键成分与核心蛋白靶点有较好的结合活性,证明了网络药理学预测结果的可靠性。研究表明,芪芝口服液可通过多成分、多靶点、多通路提升家禽的免疫功能。

基于网络药理学和体外实验探讨通关藤口服液抑制胃癌细胞增殖及联用厄洛替尼和阿帕替尼的增敏效应

目的利用网络药理学方法、分子对接及体外细胞实验探究通关藤(marsdenia tenacissima,MT)口服液抑制胃癌(gastric cancer,GC)增殖的潜在作用机制。方法通过搜索文献、检索TCMSP、Swiss Target Prediction数据库完成MT化学成分、靶点基因的收集;采用Gene Cards数据库获取疾病相关靶点;利用STRING平台和Cytoscape软件构建交集靶点的蛋白互作网络(protein-protein interaction,PPI);采用Metascape网络平台进行GO和KEGG信号通路分析;运用Auto Dock对部分活性成分与潜在靶点进行分子对接验证;采用四甲基偶氮唑盐(methyl thiazolyl tetrazolium,MTT)法检测MT口服液对GC细胞增殖的抑制作用,分别与厄洛替尼和阿帕替尼联用的协同作用、流式细胞术检测MT口服液诱导GC细胞凋亡及对胃癌细胞周期的影响;采用RT-qPCR法检测MT口服液对GC细胞周期、凋亡相关基因表达的影响;采用Western blot法验证网络药理学和分子对接筛选的靶点。结果共得到MT主要活性成分17个,胃癌关键靶点1880个,主要信号通路167条。体外实验结果表明,40~200 mg/m L的MT口服液可有效抑制GC细胞增殖(P<0.01),与厄洛替尼和与阿帕替尼联用时,具有协同效应。40~160 mg/m L的MT口服液可诱导胃癌细胞凋亡(P<0.01),阻滞细胞周期G0/G1期(P<0.01)。RT-qPCR结果显示,通关藤口服液可以上调B淋巴细胞瘤-2(B-cell lymphoma-2,Bcl-2)相关X蛋白(Bcl-2-associated X protein,BAX)的表达量,下调Bcl-2、细胞周期蛋白依赖性激酶(cyclin dependent kinase,CDK)4、CDK6(P<0.05)的表达量。Western blot结果显示MT口服液,抑制磷酸化-表皮生长因子受体(phosphorylated-epidermal growth factor receptor,p-EGFR)、磷酸化-磷脂酰肌醇-3激酶(phosphorylated-phosphatidylinositol 3-kinase,p-PI3K)、磷酸化AKT丝氨酸/苏氨酸激酶(p-AKT)、磷酸化-雷帕霉素激酶的机制靶点(phosphorylated-mechanistic target of rapamycin kinase,p-mTOR)的表达水平,降低p-EGFR/EGFR、p-PI3K/PI3K、p-AKT/AKT1、p-mTOR/m TOR比值;同时上调BAX、肿瘤蛋白p53(tumor protein p53,P53)、细胞周期蛋白依赖性激酶抑制剂(cyclin dependent kinase inhibitor 1A,CDKN1A/P21),下调Bcl-2、CDK4、CDK6(P<0.05)水平。结论MT口服液可有效抑制胃癌细胞的增殖,阻滞细胞周期并诱导胃癌细胞凋亡,并提高小分子靶向药物厄洛替尼和阿帕替尼治疗胃癌的敏感性。其机制与经调控PI3K/AKT/m TOR信号通路上调BAX、P53、P21,下调Bcl-2、CDK4、CDK6的表达,并抑制EGFR信号通路有关,实验结果验证了网络药理学和分子对接的结果。

基于UHPLC-Q-TOF/MS结合网络药理学与分子对接技术探究益心饮“异病同治”作用机制

目的:根据中医“异病同治”理论,基于UH-PLC-Q-TOF/MS技术、网络药理学、分子对接方法、细胞实验预测益心饮治疗心律失常、心力衰竭、心肌炎的核心靶点及相关信号通路。方法:采用UHPLC-Q-TOF/MS技术,对益心饮的化学成分和给药大鼠入血原形成分进行分析鉴定;并且通过TC-MSP、SwissTargetPrediction等在线数据库获取入血原形成分潜在作用靶点,利用OMIM、Genecard等数据库获取疾病靶点,做Venn图获取交集靶点,并通过STRING11.5数据库构建蛋白质-蛋白质相互作用(PPI)网络,筛选核心靶点,并选用cyto-scape3.9.0构建“疾病-成分-靶点”网络;利用DA-VID数据库对核心靶点进行GO功能和KEGG富集分析,并运用Autodock vina软件进行分子对接验证,并以H9c2细胞对潜在活性成分和靶点进行验证。结果:从益心饮中鉴定得到157个化合物;从大鼠血清中鉴定得到34个化合物,主要包括姜辣素、异甘草素、甘草次酸等化合物,得到139个交集靶点,结合PPI网络筛选得到31个核心靶点,主要含有TNF、IL-6等靶点,KEGG通路富集分析主要涉及TNF信号通路、IL-17信号通路、MAPK信号通路、PI3K-Akt信号通路等。选取TNF、IL-6靶点与主要化合物进行分子对接,对接结果均小于-5kcal/mol。体外细胞实验表明,益心饮能够通过调节TNF、IL-6发挥治疗作用。结论:益心饮主要潜在活性成分可能是异甘草素、甘草次酸、姜辣素、毛蕊异黄酮苷、丹酚酸B,主要通过作用于TNF、IL-6等靶点来调控特定的信号通路,发挥疗效。

基于网络药理学探讨舒心口服液治疗冠心病心绞痛的作用机制

目的基于网络药理学探讨舒心口服液治疗冠心病心绞痛的作用机制。方法通过TCMSP、化源网、Pubchem、SwissTarget-Prediction等平台获取舒心口服液活性成分及对应靶点,借助GeneCards数据库获取冠心病心绞痛相关靶点,对药物靶点和疾病靶点取交集,获取交集靶点。使用Cytoscape软件,构建“药物-成分-靶点”网络图,利用STRING平台获取交集蛋白相互作用信息的TSV文件,使用Cytoscape绘制蛋白质相互作用(PPI)网络,MCODE插件进行蛋白聚类分析,利用DAVID数据库与微生信进行基因本体论功能富集分析(GO)和京都基因与基因组百科全书富集分析(KEGG)及绘图。结果筛选出舒心口服液55个活性成分,包括基质分解素-1(Stromelysin-1)、转录因子p65(Transcription factor p65)、M-乙酰胆碱受体M3(Muscarinic acetylcholine receptor M3);作用于冠心病心绞痛的潜在作用靶点有160个,包括JUN、AKT1、TP53、RELA、MAPK1、TNF等;KEGG分析得到167条相关通路,主要调节肿瘤坏死因子(TNF)信号通路、磷脂酰肌醇-蛋白激酶B(PI3K-Akt)信号通路、白细胞介素-17(IL-17)信号通路、松弛素信号通路等发挥治疗冠心病心绞痛的作用。结论舒心口服液治疗冠心病心绞痛具有多靶点、多通路的特点,为舒心口服液治疗冠心病心绞痛的生物机制研究提供数据支撑,同时为进一步实验研究提供参考。

体外实验验证槲皮素治疗后发性白内障的作用机制

目的通过分子对接技术验证核心成分与核心靶点的结合能,体外实验验证槲皮素(QUE)对TGF-β2诱导的人晶状体上皮细胞(HLE-B3)的作用。方法前期通过网络药理学的方法筛选出丹红化瘀口服液(DHK)治疗后发性白内障(PCO)的核心成分槲皮素(QUE),关键靶点STAT3、JUN、TNF、TGF-β、IL-6、AKT1,利用AutoDock等软件获取核心成分与关键靶点的结合能,通过CCK-8、细胞划痕、RT-qPCR及Western blot等体外实验验证QUE对TGF-β2诱导的HLE-B3的作用。结果QUE与关键靶点STAT3、JUN、TNF、TGF-β、IL-6、AKT1有很好的结合性;体外实验结果表明QUE对TGF-β2诱导的HLE-B3细胞增殖起到抑制作用,且抑制作用呈浓度依赖性;QUE通过上调Bax和下调Bcl-2蛋白表达,发挥治疗PCO的作用。结论QUE可通过多靶点和多通路发挥治疗PCO的作用,为临床治疗提供参考。

基于网络药理学乌鳖返春口服液活性成分筛选

乌鳖返春口服液是一种结合了现代药物与食品特性的产品。本研究采用网络药理学方法,利用TCMSP数据库并结合相关文献查找其活性成分;通过SwissTargetPrediction和PharmMapper数据库预测其活性成分的可能作用靶点,在Genecards、DisGeNET和OMIM数据库检索、筛选活性成分作用的相关靶点;利用STRING数据库构建潜在作用靶点的蛋白相互作用(PPI)网络,筛选出该口服液活性成分作用的关键靶点网络,并对这些关键靶点进行GO功能和KEGG通路富集分析,确定其核心通路,以构建该口服液活性成分的作用网络。结果表明,共获得该口服液活性成分75个,基于活性成分得到384个潜在作用靶点,通过潜在作用靶点PPI网络筛选得到75个关键靶点。GO功能富集到892个条目,KEGG富集到164条通路。综合以上结果,认为该口服液活性成分作用机制可能是通过大黄蒽醌类、黄酮类和植物甾醇类化合物作用于AKT1、TNF和IL6核心靶点,以PI3KAkt、HIF-1、mTOR和FoxO为信号通路来实现。

心脉舒一号口服液治疗心脏病分子机制网络药理学研究

目的探讨心脉舒一号口服液治疗心脏病的作用机制。方法通过中药系统药理学数据库与分析平台(TCMSP)获取生地黄、天冬、麦冬、酸枣仁、柏子仁、丹参、当归、太子参、茯苓、白芍、五味子11味中药材的活性成分,并进行蛋白质靶点预测;通过GeneCards数据库获取心脏病的相关疾病靶点,通过Venny 2.1.0平台筛选出心脉舒一号口服液活性成分与心脏病的共有靶点;通过Cytoscape 3.9.1软件构建药物-活性成分-靶点网络图及活性成分-疾病靶点网络图,并分析其核心成分;通过String 11.0数据库构建蛋白-蛋白互作(PPI)网络,经Cytoscape 3.9.1软件筛选出关键靶点及核心靶点;通过Metascape数据库进行京都基因与基因组百科全书(KEGG)通路富集分析及基因生物过程(GO)功能分析;对核心成分与核心靶点进行分子对接验证。结果共收集到活性成分169种,关键靶点37个,疾病靶点1097个,活性成分-疾病共有靶点197个。核心成分为槲皮素、木犀草素、熊果酸,核心靶点为环氧合酶2(PTGS2)、钠电压门控通道α亚基5(SCN5A)、溶质载体家族6成员2(SLC6A2)。共获得KEGG信号通路17条,主要涉及蛋白解旋酶癌、人类巨细胞病毒感染、晚期糖基化终末产物-晚期糖基化终末产物多配体受体(AGE-RAGE)信号通路等;GO功能分析条目1205条,包括生物学过程(BP)944条、分子功能(MF)146条和细胞组分(CC)115条,主要涉及正调控血管内皮细胞迁移、老化、炎性反应、转录阳性调控-DNA模板化、基因表达的负调控、正调控细胞增殖等。分子对接结果显示,核心成分与其核心靶点均有较好的结合活性。结论心脉舒一号口服液可通过多成分、多靶点、多通路治疗心脏病。

基于网络药理学和分子对接技术揭示血府逐瘀口服液抗血栓的活性成分和作用机制

为揭示血府逐瘀口服液治疗血栓类疾病的机理,挖掘血府逐瘀口服液中抗血栓有效成分。通过TCMSP数据库检索血府逐瘀口服液中的化学成分或成分靶点;利用STRING数据库构建蛋白质-蛋白质相互作用(PPI)网络得到核心靶点;通过Cytoscape构建“成分-靶点”网络图,并进行拓扑分析和对核心靶点进行GO和KEGG富集分析,预测血府逐瘀口服液抗血栓的作用机制;根据Degree值排名将关键的成分和作用靶点进行分子对接。通过网络拓扑分析筛选得到81个核心靶点,如TNF、ALB、AKT1等;GO富集分析中生物过程(BP)共304条,分子功能(MF)共72条,细胞组成(CC)共41条,通路富集得到凝血级联反应、TNF等80条信号通路;分子对接结果显示,Sainfuran、Xambioona和7-Methoxy-2-methyl isoflavone与靶点蛋白ESR1、F2、IL-2、KDR、MET、MMP3均有较好的亲和力。该研究为血府逐瘀口服液在抗血栓应用方面提供参考。

基于抗炎和促红细胞生成作用的九味补血口服液改善血虚证小鼠造血功能的机制研究

为探讨九味补血口服液改善血虚证小鼠造血功能的作用及机制,采用超高效液相色谱串联四级杆飞行时间质谱(UPLC-Q-TOF-MS/MS)技术鉴定九味补血口服液中的化学成分;利用蛋白互作和网络分析手段筛选九味补血口服液治疗贫血的关键网络靶点,并对关键网络靶点进行基因本体(GO)分析、京都基因与基因组百科全书(KEGG)通路富集分析;同时采用分子对接技术将潜在活性成分与关键网络靶点对接,评估活性分子与靶点的结合能力。在动物实验中,采用环磷酰胺建立小鼠血虚模型,设立对照组和模型组,复方阿胶浆口服液组(30 mL/kg)及九味补血口服液高、中、低剂量组(10.0、5.0、2.5 mL/kg),每天灌胃给药1次,连续32 d。检测全血中红细胞(RBC)、血红蛋白(HGB)和红细胞压积(HCT)水平,并利用qRT-PCR法和免疫印迹法对关键靶点及通路在血虚证小鼠肾脏的表达进行验证。结果显示,在91种鉴定的九味补血口服液化学成分中,满足口服生物利用度(OB)≥30%和药物相似性(DL)≥0.18的条件,且具有相应靶点的潜在活性成分共计15个。分析九味补血口服液成分-血虚疾病靶点网络,获得九味补血口服液治疗贫血的关键靶点15个,主要富集于核因子-κB(NF-κB)信号通路(NF-κB signaling pathway)、代谢通路(Metabolic pathway)、缺氧诱导因子-1(HIF-1)信号通路(HIF-1 signaling pathway)、Toll样受体信号(Toll-like receptor signaling pathway)通路等。分子对接结果显示,甘草苷、橙皮素与Toll样受体4(TLR4)具有强烈的结合能力。动物实验结果显示,与对照组相比,血虚证小鼠RBC、HGB和HCT水平明显降低,TLR4/NF-κB p65介导的炎性信号增强,缺氧诱导因子-1α(HIF-1α)/促红细胞生成素(Erythropoietin,EPO)介导的促红细胞生成信号受抑制(P<0.05或P<0.01)。与模型组相比,九味补血口服液各剂量组可显著升高血虚证小鼠RBC、HGB和HCT水平,不同程度地逆转TLR4、NF-κB p65变化,并上调HIF-1α、EPO mRNA或蛋白的表达水平(P<0.05或P<0.01)。因此,九味补血口服液可以改善血虚证小鼠的贫血状态,其机制可能与促进肾脏中HIF-1α/EPO介导的促红细胞生成作用、减少TLR4/NF-κB p65介导的肾脏炎症反应相关。

Network pharmacology and in vitro study of Fengreqing oral liquid(风热清口服液) in the intervention of wind-heat pattern

OBJECTIVE:To investigate the underlying mechanism of the effect of Fengreqing oral liquid(风热清口服液,FOL)on wind-heat pattern(WHP).METHODS:In this study,we predicted the potential targets of FOL via the approach of network pharmacology and verified it by in vitro inflammation model.In the network pharmacology part,two strategies,namely the direct target search and the indirect one,were used to collect the target sets of FOL in WHP treatment.The enrichment analysis was carried out by David database and Clue Go plug-in in Cytoscape.Furthermore,the potential targets were mapped in the candidate pathways.In the verification experiment section,in vitro model of lipopolysaccharide(LPS)induced RAW 264.7 was used to confirm the predictive results in the network pharmacology part.RESULTS:Through the two screening strategies,a total of 141 non-repetitive intervention targets of FOL on WHP were obtained.Gene ontology(GO)and Kyoto Encyclopedia of Genes and Genomes(KEGG)enrichment analysis showed that the intervention effect was mainly focused on the anti-inflammatory effect,and the Toll-like receptor signaling pathway was one of the most critical regulatory pathways.Further mapping analysis showed that phosphatidylinositol 3-kinase(PI3 K)-protein kinase B(AKT)signaling transfer might be the key part of regulating the concentration of inflammation mediators of FOL in the Toll-like receptor signaling pathway.In vitro experiment showed that FOL significantly reduced the levels of NO,IL-1,IL-6,and TNF-αproduced by RAW264.7 induced by LPS.Further immunofluorescence found that this effect is related to the regulation of PI3 K-AKT pathway activity by FOL.CONCLUSION:FOL can intervene in WHP by regulating the content of inflammatory mediators via the PI3 K-AKT pathway.

基于网络药理学和分子对接探讨金银花口服液防治新型冠状病毒感染的作用机制

利用网络药理学和分子对接探讨金银花口服液防治新型冠状病毒感染的分子机制。通过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)检索药物的化学成分,借助Swiss Target Prediction数据获取药物成分对应的相关靶点;在GeneCards、CTD、TTD、DrugBank数据库中检索疾病的相关靶点;利用Venny 2.1.0在线软件获取药物与疾病的共同靶点;由Cytoscape 3.8.2绘制药物-成分-靶点-疾病网络;String数据库构建蛋白互作网络;借助David数据库进行基因本体论(gene ontology,GO)功能富集和京都基因和基因组百科全书(kyoto encyclopedia of genes and genomes,KEGG)通路富集分析;运用AutoDock软件对成分和靶点进行分子对接验证。结果发现,金银花口服液中16个有效成分通过调控88个靶点和108条通路对新型冠状病毒感染产生作用,伞房花耳草素、金圣草素、木犀草素、亚油酸乙酯、山奈酚、槲皮素等成分可以通过AKT1、SRC、EGFR、HIF1A、PPARG、MMP9、ESR1、MAPK1、PTGS2、MMP2等关键靶点调控HIF-1、化学致癌-受体激活、内分泌抵抗、粘着斑、PI3K-Akt、VEGF、松弛素、ErbB、丙型肝炎、乙型肝炎、Rap1等信号通路来治疗新型冠状病毒感染。分子对接结果显示,靶点蛋白与活性成分具有较好的结合能力。结果表明,金银花口服液可以通过多成分、多靶点、多途径参与新型冠状病毒感染的防治,这为其临床应用及深入研究防治新型冠状病毒感染的机制提供了理论依据。

基于网络药理学和分子对接技术探讨脑血疏口服液治疗缺血性卒中的作用机制

目的采用网络药理学和分子对接技术探索脑血疏口服液治疗缺血性卒中的药效物质基础与潜在作用机制。方法基于脑血疏口服液与缺血性卒中的共同靶点构建“单味药-活性成分-作用靶点”网络;通过String数据库构建蛋白互作(PPI)网络;Metascape数据库进行GO功能和KEGG通路注释与富集分析;分子对接分析预测关键的活性成分与靶点的结合互作。结果“单味药-活性成分-作用靶点”网络中获得脑血疏口服液治疗缺血性卒中的可能活性成分29个,作用靶点67个,其中关键活性成分包括山柰酚、β-胡萝卜苷、胡萝卜苷等;PPI网络分析获得AKT1、PTGS2、EGFR、SRC等关键靶点,其中山柰酚与以上四个关键靶点均有较高的亲和力;脑血疏口服液治疗缺血性卒中的机制可能主要涉及激酶活性、炎症反应、氧化应激应答、免疫系统过程等多种分子功能和生物过程,以及EGFR酪氨酸激酶抑制剂抗性、PI3KAkt信号通路等165个通路。结论脑血疏口服液通过多成分、多靶点、多通路和功能的复杂网络调控炎症反应、血管损伤、神经元损伤或凋亡及脑损伤等防治缺血性卒中,为其后续深入药理学研究奠定了基础。

基于网络药理学和分子对接探究归芪益母口服液治疗猪气虚血瘀证的作用机制

【目的】运用网络药理学和分子对接技术探讨归芪益母口服液治疗猪气虚血瘀证的有效活性成分及其作用机理。【方法】通过中药系统药理学数据库与分析平台(TCMSP)、中医药综合数据库(TCMID)获取归芪益母口服液主要活性成分及其靶点,通过GeneCards、OMIM和Malacards数据库获得母猪气虚血瘀证的作用靶点。使用Cytoscape 3.9.0软件构建药物-成分-靶点网络图,利用STRING数据库和Cytoscape 3.9.0软件构建靶点蛋白-蛋白互作(PPI)网络图,采用DAVID数据库对共同作用靶点进行GO功能和KEGG信号通路富集分析。通过Autodock Tools 1.5.7软件对其核心成分与核心靶点进行分子对接。【结果】归芪益母口服液筛选得到主要的有效活性成分为槲皮素、山奈酚和异鼠李素,其对应76个关键作用靶点,6335个疾病靶点,41个交集靶点;PPI网络显示肿瘤坏死因子(TNF)、白介素-6(IL6)和IL10为关键靶点。GO功能富集分析共筛选出12条目,其中生物过程7条目,包括免疫应答、STAT蛋白酪氨酸磷酸化的正向调节、对糖皮质激素的反应等;细胞组分2条目,包括细胞外组分和细胞外空隙;分子功能3条目,包括细胞因子、生长因子活性和IL2受体结合。KEGG信号通路富集分析显示,归芪益母口服液主要通过细胞因子受体互作、JAK-STAT信号通路、PI3K-Akt信号通路发挥作用。分子对接结果显示,槲皮素、山奈酚、异鼠李素等核心成分与母猪气虚血瘀证关键靶点的结合能均<0 kJ/mol。【结论】归芪益母口服液可能通过槲皮素、山奈酚、异鼠李素等主要活性成分作用于核糖体结合蛋白(RPN1)、胰核糖核酸酶(RNASE1)、血管生成蛋白抑制因子1(RNH1)等靶点,通过参与细胞因子受体互作、JAK-STAT信号通路、PI3K-Akt信号通路等治疗猪气虚血瘀证。