Targeting colorectal cancer with Herba Patriniae and Coix seed:Network pharmacology,molecular docking,and in vitro validation

BACKGROUND Herba Patriniae and Coix seed(HC)constitute a widely utilized drug combination in the clinical management of colorectal cancer(CRC)that is known for its diuretic,anti-inflammatory,and swelling-reducing properties.Although its efficacy has been demonstrated in a clinical setting,the active compounds and their mechanisms of action in CRC treatment remain to be fully elucidated.AIM To identify the active,CRC-targeting components of HC and to elucidate the mechanisms of action involved.METHODS Active HC components were identified and screened using databases.Targets for each component were predicted.CRC-related targets were obtained from human gene databases.Interaction targets between HC and CRC were identified.A“drug-ingredient-target”network was created to identify the core components and targets involved.Gene Ontology(GO)and Kyoto Encyclopedia of Genes and Genomes(KEGG)analyses were conducted to elucidate the key pathways involved.Molecular docking between core targets and key components was executed.In vitro experiments validated core monomers.RESULTS Nineteen active components of HC were identified,with acacetin as the primary active compound.The predictive analysis identified 454 targets of the active compounds in HC.Intersection mapping with 2685 CRC-related targets yielded 171 intervention targets,including 30 core targets.GO and KEGG analyses indicated that HC may influence the phosphoinositide 3-kinase(PI3K)/Akt signaling pathway.Molecular docking showed that acacetin exhibited an optimal interaction with AKT1,identifying PI3K,AKT,and P53 as key genes likely targeted by HC during CRC treatment.Acacetin inhibited HT-29 cell proliferation and migration,as well as promoted apoptosis,in vitro.Western blotting analysis revealed increased p53 and cleaved caspase-3 expression and decreased levels of p-PI3K,p-Akt,and survivin,which likely contributed to CRC apoptosis.CONCLUSION Acacetin,the principal active compound in the HC pair,inhibited the proliferation and migration of HT-29 cells and promoted apoptosis through the PI3K/Akt/p53 signaling pathway.

Exploring the effect and mechanisms of Epimedium on diabetic testicular injury using network pharmacology,molecular docking,and cell experiments

Background:Studying the potential targets and mechanisms of Epimedium for anti-diabetic testicular injury using network pharmacology,molecular docking,and cell experiments.Methods:Acquisition of major components and targets of Epimedium was based on TCMSP,TCMID,and Symmap databases and retrieval of diabetic testicular injury targets by OMIM,GeneCards,Pharmgkb,and Drugbank databases.Intersecting targets were obtained from the Venny 2.1.0 database and input SRTING data to construct a protein-protein interaction(PPI)network,and key targets were screened in Cytoscape 3.8.0 software.Then the Gene Ontology(GO)and Kyoto Encyclopedia of Genes and Genomes(KEGG)enrichment analyses of intersecting targets were conducted through the DAVID database.Further,AutoDock software was used to verify docking between the main components and the core target proteins.In addition,a Cell Counting Kit-8(CCK-8)assay was used to determine the survival effect of quercetin,the main component of Epimedium,on TM4 sertoli-like cells exposed to palmitic acid(PA).Results:Quercetin,kaempferol and luteolin in epimedium were identified as the main components in the treatment of diabetic testicular injury.It has core target proteins including MMP9,AKT1,and TNF.The biological process mainly involves the regulation of the apoptotic signaling pathway.The key pathways of KEGG are the AGE-RAGE signaling pathway in diabetic complications,PI3K-Akt and MAPK signaling pathway.Molecular docking results showed that quercetin had the strongest binding ability to MMP9.Also,PA-challenged cells had a lower survival rate,which was alleviated by the administration of quercetin.Conclusion:Our findings suggest that Epimedium attenuates diabetes mellitus(DM)-induced testicular injury through AGE-RAGE,PI3K-Akt and MAPK signaling pathway.These insights offer a potential therapeutic strategy for managing DM-induced testicular injury,will be the basis for future clinical research.

Study on mechanism of two related Chinese herbs A.officinarum-Pogostemon in treatment of delayed emptying in diabetic gastroparesis based on network pharmacology

Objective:To explore the potential active components,therapeutic targets and critical path of Alpinia officinarum and Pogostemonis Herba in the treatment of diabetic gastroparesis(DGP)by using network pharmacology.Methods:The main chemical components and corresponding targets genes of A.officinarum-Pogostemonis Herba were screened through TCMSP database retrieval[oral bioavailability(OB)≥30%and drug like(DL)≥0.18].Tgenes of diabetic gastroparesis were screened by the Human Gene Database(GeneCards),and Venny 2.1 software was used to obtained common targets for the active ingredients of A.officinarum-Pogostemonis Herba and DGP.Then,the protein-protein interaction(PPI)network of the common targets was constructed by STRING database and analyzed to performed the core targets.GO function and KEGG pathway enrichment analysis of the common target genes were obtained by using ClusterProfiler R package.Finally,the network diagram of"active ingredient-pathway-target"was used to establish by Cytoscape 3.8 software.Results:Totally 23 ingredients of A.officinarum-Pogostemonis Herba,97 active ingredients targets and 533 DGP related targets,including 46 common targets were selected.The common targets were mainly enriched in the cell constituents such as the nuclear chromatin and mitochondria outer membrane,involved in the biological processes as oxidative stress,apoptosis signal regulation,and molecular functions as enzyme binding,protein phosphatase binding,and cytokine activity.They were also concentrated in the signal pathways such as PI3K/Akt,HIF-1 and MAPK.The network of“active ingredients-targets-pathways”indicated the active components such as quercetin,kaempferol and galangin in A.officinarum-Pogostemonis Herba played an anti-delayed gastric emptying in diabetic gastroparesis by acting on PTGS2,NOS2,BCL2,IL6,VEGFA and other targets to jointly regulate PI3K-Akt,HIF-1 and MAPK pathways.Conclusion:This study initially reveals that the combined treatment of A.officinarum-Pogostemonis Herba for delayed gastric emptying in diabetic gastroparesis is a complex process with multi-components,multi-targets and multi-pathways,and provides a new idea for followup researches.

Network Pharmacology Analysis of Epimedium brevicornu(Yinyanghuo)for Treatment of Ovarian Cancer

[Objectives]To elucidate potential targets and mechanisms of action of Epimedium brevicornu in treating ovarian cancer.[Methods]The Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform was used to screen active components of E.brevicornu for disease control and prevention,and potential targets were collected from the DisGeNET database.These sets of bioactive and targets were analyzed using Ingenuity Pathway Analysis(IPA)to predict molecular networks affected by E.brevicornu in ovarian cancer.Venny 2.1.0 software was used to screen for proteins affected by interactions between disease and active components,which were input into the STRING 11.0 platform to construct a protein-protein interaction network.Then IPA and STRING were used to analyze common targets which were obtained from the two data analysis platform.[Results]A total of 23 major active components of E.brevicornu and 200 potential human targets were screened.IPA analysis identified 363 pathways and 24 networks shared between the set of predicted Yinyanghuo targets and ovarian cancer-associated proteins.These pathways are involved mainly in molecular mechanisms of cancer,glucocorticoid receptor signaling pathways,pancreatic adenocarcinoma signaling pathways,aryl hydrocarbon receptor signaling pathways,and macrophage function.The 24 networks have been implicated mainly in cancer,endocrine system disorders,body damage and abnormality,cell growth and proliferation,connective tissue development and function,tissue development,and other biological functions.IPA and STRING combined analysis suggested that AKT1,CASP3,JUN,FOS and CCND1 are the most likely targets of Yinyanghuo in treating ovarian cancer.[Conclusions]Our network pharmacology analysis identified several pathways that Yinyanghuo may influence to reduce ovarian cancer risk;in particular,it identified specific protein targets,including AKT1,CASP3,JUN,FOS and CCND1.

Pharmacology of Active Components of Glechomae Herba in the Treatment of Bladder Cancer

[Objectives]To explore the action mechanism of Glechomae Herba in the treatment of bladder cancer through network pharmacology.[Methods]TCMSP database retrieval and literature text mining were used to find out the main chemical active components and their targets,and the"component-target"network map was constructed by Cytoscape 3.7.0 software.The CTD database was used to select targets related to bladder cancer,and the interaction between the targets of the main active components of Glechomae Herba and the targets related to bladder cancer was taken.A total of 87 targets were obtained,then imported into the STRING database to obtain PPI network,and the topological analysis of the network was conducted to find out the key targets.The key targets were introduced into the DAVID database,and the biological process and KEGG(Kyoto Encyclopedia of Genes and Genomes)pathway of the key targets in the treatment of bladder cancer were obtained.[Results]Active compound components interfere with the occurrence and development of bladder cancer possibly through bladder cancer signaling pathway,proteoglycan in cancer signaling pathway,HIF-1 signaling pathway,tumor necrosis factor signaling pathway,PI3K-Akt signaling pathway,toll-like receptor signaling pathway,node-like receptor signaling pathway,and MAPK signaling pathway,etc.[Conclusions]The pharmacological mechanism of Glechomae Herba in the treatment of bladder cancer may be realized through multiple components,multiple targets and multiple pathways.In conclusion,Glechomae Herba has certain medicinal value.

A network pharmacology approach combined with animal experiment to investigate the blood enriching effect of Gei herba

Background:To explore active components of Lanbuzheng(Gei herba)and its underlying complex mechanism in treating blood deficiency induced by chemotherapy drug based on network pharmacology and mice experimental validation.Methods:Active components of Lanbuzheng(Gei herba)were screened by Lipinski’s rule of five.Targets acted with active components were predicted by PharmMapper database,and targets whose function associated with blood deficiency were screened by Therapeutic Target Database and UniProt.The networks of component-target and target-pathway were constructed by Cytoscape.The levels of peripheral blood and organ indexes were detected in the animal experiments.Results:One hundred and seventy-three components of Lanbuzheng(Gei herba)were collected,and 60 active components were screened according to the rule of five.According to the degree value of compounds,the top 5 compounds were docosyl trans ferulate,C32 decursin,agrimonolide 6-O-β-D-glucoside,degree=11,173-ethoxyphaeophorbide,and eugenol.Finally,59 targets associated with blood deficiency were obtained and the top 5 targets were MAPK14,TTR,CDK2,AKR1B1 and AR.Based on the interaction network of componenttarget and target-pathway,it’s found that 60 active components could act with 59 targets and 44 pathways for treating blood deficiency.And then,the mice experiments showed that Lanbuzheng(Gei herba)could enrich blood by increasing the levels of red blood cell,white blood cell,hemoglobin,red blood cell specific volume and platelet,and the indexes of liver,thymus and spleen,which validated the treating effect of Lanbuzheng(Gei herba).Conclusion:In this study,a network pharmacology approach and animal experiments were established to explore the nourishing blood effect of Lanbuzheng(Gei herba).The results demonstrated that Lanbuzheng(Gei herba)could improve blood deficiency and provide a theoretical basis for the further research on the in-depth mechanism of Lanbuzheng(Gei herba).

Study on the mechanism of action of Ephedra Herba Decoction against influenza A virus based on network pharmacology

Objective To predict the main bioactive components and potential mechanisms of Ephedra Herba Decoction(Chinese Pinyin abbreviated as MHT)in treating influenza A virus(IAV)based on network pharmacology.Methods Multiple online databases were used to search and screen out the active components from MHT,the related targets of active components of MHT and the genes related to IAV.Search the corresponding genes name of target through UniProt database.Cytoscape 3.7.2 was used to construct the drug-component-target network diagram.Venn diagram was used to screen the intersection genes of the active components corresponding to the target and disease-related genes,and the intersection genes were imported into the STRING Database Online platform to obtain the protein-protein interaction(PPI)network.Then,the PPI network was imported into Cytoscape 3.7.2 to obtain the core targets.Finally,Wei Sheng Xing(http://www.bioinformatics.com.cn/)was used to do GO function enrichment analysis and KEGG signaling pathway enrichment analysis for the intersection genes,and the results of GO and KEGG were visualized.Results A total of 116 active components and 253 potential targets were screened from MHT.Quercetin,Kaempferol,and Luteolin are the main active components,and AKT1,TNF,TP53,IL6,and JUN are the core targets of 253 potential targets.There are 2906 targets of influenza A,including 121 intersection genes.Enrichment analysis showed that there were 131 entries of molecular function,89 entries of cell component,1645 entries of biological process and 195 entries of signaling pathway.PI3K-Akt,MAPK,and JAK-STAT were the main signaling pathways.Conclusion MHT plays an important role in the prevention and treatment of influenza A by acting on multiple targets and multiple signaling pathways.

Molecular mechanism of Herba Eupatorii in treating COVID-19 based on network pharmacology and molecular docking

Objective:To explore the therapeutic target and molecular mechanism of Herba Eupatorii in the intervention of COVID-19(coronavirus disease 2019)by network pharmacology.Methods:TCMSP(Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform)and TCMIP V2.0(Integrative Pharmacology-based Research Platform of Traditional Chinese Medicine)databases were used to search the active ingredients and corresponding drug targets of Herba Eupatorii.Related targets of COVID-19 were searched in Genecards,pharmGKB,CTD,Drugbank and TTD databases.After the intersection targets were selected using VENNY 2.1 online platform,the PPI(protein-protein interaction)network was downloaded into STRING database,and the data were analyzed and sorted out using Cytoscape software to obtain the potential key targets for the treatment of COVID-19 by Herba Eupatorii.At the same time,using the data of active ingredients and intersection targets,a network of"TCM-active ingredients-key targets"was constructed in Cytoscape software to screen out chemical molecules with potential therapeutic effects.GO(Gene Ontology)functional enrichment analysis and KEGG(Kyoto Encyclopedia of Genes)pathway enrichment analysis of key target proteins were performed by R software.AutoDock Vina program was used for molecular docking of the top 5 active ingredients and key targets to calculate the minimum binding energy.Results:There were 26 active ingredients,160 targets,and 1969 pathogenic genes of COVID-19,among which 59 genes were intersection targets of drugs and diseases.After PPI network screening,the key target proteins were AKT1(RAC-alpha serine/threonine-protein kinase),JUN(transcription factor AP-1),TP53(cellular tumor antigen p53),ACTB(actin beta)and EGFR(epidermal growth factor receptor).Through the network of"TCM-Active Ingredients-Key Targets",Luteolin,Eupatolin,Stigmasterol,Eupatoriopicrin and Dammaradienyl acetate were identified as the active ingredients with potential therapeutic effects in the treatment of COVID-19.After R software was used for GO enrichment analysis,1978 GO items were obtained(P<0.05),including 1870 BP items,26 CC items and 82 MF items.149 pathways were obtained by KEGG enrichment analysis(P<0.05).It mainly involves IL-17(interleukin-17)signaling pathway,TNF(tumor necrosis factor)signaling pathway,C-type lectin receptor signaling pathway,PI3K-Akt(phosphatidylinositol 3 kinase-protein kinase B)signaling pathway,and T Cell receptor signaling pathway,etc.The molecular docking results showed that the active ingredients had good binding activity with key targets.Conclusion:Through the potential chemical constituents of Luteolin,Eupatolin,Stigmasterol,Eupatoriopicrin and Dammaradienyl acetate,Herba Eupatorii may act on AKT1,JUN,TP53,ACTB,EGFR and other targets.Involvement in IL-17 signaling pathway,TNF signaling pathway,C-Type Lectin receptor signaling pathway,PI3K-Akt signaling pathway,T Cell receptor signaling pathway and other pathways play an anti-inflammatory and antiviral roles in intervening in the occurrence and development of COVID-19.

Research progress on chemical constituents in Sigesbeckiae Herba and their pharmacological activities

Sigesbeckiae Herba,a dry whole plant of Sigesbeckia glabrescens Makino,Sigesbeckia pubescens Makino and Sigesbeckia orientalis L.,is a traditional Chinese medicine used to dispel dampness,dredge meridians and collaterals,clear heat and detoxicate.Its chemical components are complex and abundant,including mainly diterpenoids,sesquiterpenoids and flavonoids.Among them,diterpenoids are the main active components.Modern pharmacological studies have found that the compounds isolated from Sigesbeckiae Herba have anti-inflammatory,anti-tumor,antithrombus and other pharmacological activities.This review systematically summarized the chemical components identified from Sigesbeckiae Herba and their pharmacological activities by searching Chinese and English databases,in order to provide references for the further development of Sigesbeckiae Herba and the improvement of its quality standards.

Investigating the molecular mechanism of Chelidonii Herba against liver cancer using network pharmacology and molecular docking validation

Background:The molecular mechanism of Chelidonii Herba in treating hepatocellular carcinoma was investigated using network pharmacology and molecular docking validation.Methods:The main active components of Chelidonii Herba were screened using the Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform database,and the targets of these active ingredients were identified using the SwissTargetPrediction platform.Targets related to liver cancer were sourced from GeneCards,Therapeutic Targets Database,and Online Mendelian Inheritance in Man databases.Intersection targets between the active components of Chelidonii Herba and liver cancer were determined using the jvenn online platform.The protein interaction network was analyzed via STRING database and visualized using Cytoscape 3.9.1.Core targets were identified and further analyzed within the protein interaction network.Gene Ontology and Kyoto Encyclopedia of Genes and Genomes enrichment analyses were conducted for the intersection targets using the DAVID database to correlate gene functions.Sankey bubble diagrams for Gene Ontology enrichment analysis and circular diagrams for Kyoto Encyclopedia of Genes and Genomes pathway enrichment analysis were generated using CNSknowall and SangerBox online platforms.Molecular docking and visualization were performed using AutoDockTools 1.5.7 and PyMOL 2.5.7 software,respectively.Overall survival and pan-cancer analysis of core targets were conducted using the GEPIA2 online platform.Results:Twelve active components of Chelidonii Herba were identified through screening.A total of 103 intersection targets and 12 core targets were found between these active constituents of Chelidonii Herba and liver cancer.Chelidonii Herba may exert its effects on liver cancer through these 12 core targets.Several signaling pathways are implicated,including chemical carcinogen-receptor activation,endocrine resistance,HIF-1 signaling pathway,and proteoglycans in cancer.Conclusion:Chelidonii Herba potentially intervenes in cancer-related signaling pathways for treating liver cancer by targeting AKT1,EGFR,and ERBB2.This action is facilitated by active ingredients such as(S)-chrysocorydaline,dihydrochelidonorubin,cryptopine,and oxysanguinarine.Chelidonii Herba may address liver cancer through a mechanism involving multiple components,targets,and pathways.

基于网络药理学和分子对接技术探讨三七-淫羊藿治疗股骨头坏死的作用机制

目的运用网络药理学和分子对接技术探究三七-淫羊藿治疗股骨头坏死(ONFH)的作用机制。方法通过中药系统药理学数据库与分析平台筛选三七-淫羊藿的活性成分及其作用靶点,利用GeneCards®数据库、OMIM®数据库筛选ONFH的相关靶点,取交集后获得三七-淫羊藿治疗ONFH的潜在作用靶点。通过Cytoscape软件构建药物-活性成分-作用靶点网络,筛选关键活性成分。通过STRING数据库和Cytoscape软件构建蛋白-蛋白相互作用网络,筛选核心靶点。使用DAVID数据库对潜在作用靶点进行功能富集分析和通路富集分析。最后,对关键活性成分与核心靶点蛋白进行分子对接验证。结果最终获得三七-淫羊藿活性成分30个及其作用靶点214个,ONFH的相关靶点2112个。取交集后得到三七-淫羊藿治疗ONFH的潜在作用靶点131个。槲皮素、木犀草素、山柰酚、脱水淫羊藿素为三七-淫羊藿治疗ONFH的关键活性成分,蛋白激酶B1、血管内皮生长因子A、原癌基因c-Jun、肿瘤蛋白p53、白细胞介素1β为三七-淫羊藿治疗ONFH的核心靶点。三七-淫羊藿治疗ONFH的潜在作用靶点主要涉及对脂多糖的反应、对外来刺激的反应、细胞缺氧反应等生物过程,以及白细胞介素17信号通路、丝裂原活化蛋白激酶信号通路、糖尿病并发症中的晚期糖基化终末产物-晚期糖基化终末产物受体信号通路、磷脂酰肌醇3-激酶/蛋白激酶B信号通路等信号通路。分子对接结果显示,关键活性成分与核心靶点蛋白均具有较好的结合活性(结合能≤-5 kcal/mol)。结论三七-淫羊藿治疗ONFH的作用机制可能与调控免疫-炎症反应、调控破骨细胞与成骨细胞的凋亡和分化、改善血运等有关。

淫羊藿治疗骨质疏松骨折的网络药理学分析

目的利用网络药理学原理探讨淫羊藿治疗骨质疏松骨折的分子作用机制。方法利用中药系统药理学分析平台(TCMSP数据库)筛选淫羊藿的主要活性成分和作用靶点,应用GeneCards数据库、OMIM数据库、PharmGkb数据库、TTD数据库、DrugBank数据库预测骨质疏松骨折的相关靶点,制作维恩图获取淫羊藿与骨质疏松骨折的共同靶点,运用Cytoscape软件绘制中药-成分-疾病-靶点调控网络,利用STRING数据库进行蛋白相互作用网络的构建与分析。最后,使用R语言和Bioconductor平台进行基因本体富集分析(GO)和基因相互作用通路分析(KEGG)。结果淫羊藿主要是通过23种活性成分、121个作用靶点及155个生物学通路发挥作用治疗骨质疏松骨折。其中主要活性成分为槲皮素、山奈酚和木犀草素;主要核心靶点有AKT1、TP53、Jun、HSP90AA1和CASP3等;关键通路为脂质和动脉粥样硬化信号通路。结论通过网络药理学揭示了淫羊藿通过多成分、多靶点、多通路治疗骨质疏松骨折的作用特点,为进一步的研究和实验工作提供了基础。

补骨脂-淫羊藿对类风湿关节炎抗炎机制的分子对接分析:动物实验验证

背景:临床上补骨脂-淫羊藿治疗类风湿关节炎疗效明显,但两者所含有效成分复杂,在分子水平上治疗类风湿关节炎的作用机制仍不明确。目的:基于网络药理学和分子对接技术建立胶原诱导型关节炎模型,验证补骨脂-淫羊藿治疗类风湿关节炎可能的作用靶点及通路,为以补骨脂-淫羊藿为主的临床方剂使用提供可靠的实验依据。方法:借助中医药研究平台、中医百科全书和上海有机所的中药与化学成分数据库等检索并筛选有效成分,从PubChem平台获取3D分子式,通过PharmMapper和SwissTargetPrediction平台进行靶标预测;结合DrugBank、GeneCards、OMIM等基因数据库完成类风湿关节炎的疾病靶点获取,经Uniport数据库校准靶标,借助VENNY 2.1获取补骨脂-淫羊藿与疾病交集靶点并绘制韦恩图;采用STRING平台构建蛋白质互作网络图;使用Metascape平台进行基因本体论功能分析及京都基因与基因组百科全书分析,进行数据可视化,利用Cytoscape 3.9.0构建中药-成分-靶点-疾病-通路四重网络模型;运用AutoDock-Vina软件将主要有效成分与核心靶点进行分子对接验证,探索最佳结合靶点。建立Ⅱ型胶原+佐剂诱导型关节大鼠模型,用补骨脂-淫羊藿干预21 d后,观察其对相关通路靶点及炎性细胞因子的影响。结果与结论:①筛选补骨脂与淫羊藿活性成分28个,与类风湿关节炎交集靶点共288个,主要成分有异补骨脂素、补骨脂定、淫羊藿苷等;交集靶点主要有丝氨酸/苏氨酸蛋白激酶1(AKT1)、肿瘤坏死因子、血管内皮生长因子A等;②基因本体论分析获得生物过程2232条,主要与丝氨酸蛋白磷酸化、AKT正调控、活性氧代谢过程等功能有关;③京都基因与基因组百科全书富集分析结果202条,主要有PI3K/AKT信号通路和表皮生长因子受体信号通路等,可能通过调节滑膜细胞凋亡与增殖、抑制炎性因子等发挥治疗作用;④分子对接结果表明补骨脂-淫羊藿主要与AKT1及雌激素受体转录因子1结合活性最强,并形成稳定结构,与PI3K/AKT等凋亡增殖、炎性介导等调控信号通路密切相关;⑤补骨脂-淫羊藿可降低胶原诱导型关节炎大鼠模型血清中白细胞介素1β、白细胞介素6、肿瘤坏死因子α的表达;⑥补骨脂-淫羊藿可调低胶原诱导型关节炎大鼠模型关节滑膜中p-PI3K、p-AKT、p-FOXO1蛋白的表达;⑦结果证明,补骨脂-淫羊藿可能经PI3K/AKT/FOXO1信号通路抑制关节滑膜细胞增殖和抑制炎性因子表达等发挥治疗作用,这可能与类风湿关节炎关节炎症和骨破坏的发生密切相关,同时为临床的合理使用及新药开发提供了参考依据。

基于多组学与网络药理学探究淫羊藿对后备母猪发情的作用

旨在揭示淫羊藿对后备母猪发情的作用及其机理。本试验选择210~220日龄,体重(99.547±1.987)kg,发育成熟且符合配种条件的后备待配二元母猪32头,随机分为对照组和试验组,每组16头母猪,每个重复1头母猪。对照组饲喂基础日粮,试验组在饲喂基础日粮的基础上补充淫羊藿粗提物50 mg·d^(-1)。试验饲喂28 d。结果表明,试验组母猪发情提前,血清FSH、LH和E_2显著增加(P<0.05)。卵巢转录组结果表明,检测出477个上调差异mRNAs,754个下调差异mRNAs。GO富集分析发现,差异的mRNA主要富集在运输囊泡、脂肪细胞分化、节律行为、发情周期等过程;KEGG富集分析发现,差异的mRNA主要富集在紧密连接、碳水化合物代谢、刺猬信号通路、GnRH分泌和PI3K-AKT信号通路等信号通路。卵巢代谢组结果表明共有1616个代谢物上调,1254个代谢物下调。KEGG富集分析表明,差异代谢物主要富集在α-亚麻酸代谢、ATP转运蛋白、亚油酸代谢、β-丙氨酸代谢、氨基苯甲酸盐降解、生物素代谢等通路。网络药理学分析结果表明淫羊藿作用卵巢的潜在作用靶点共161个。PPI互作网络得到degree前10的蛋白作为核心靶点,根据得分依次为TP53、SRC、AKT1、CCND1、TNF、ESR1、EP300、ERBB2、JAK2、PARP1。GO分析结果表明,淫羊藿作用卵巢的靶点主要参与了蛋白磷酸化、MAPK级联反应的正调控、生物节律、基因表达的正调控、细胞凋亡过程的负调控、细胞内钙离子浓度的正调控、RNA聚合酶II启动子转录的正调控等生物学过程。KEGG分析结果显示,靶点蛋白信号通路富集在PI3K-Akt信号通路、细胞周期、细胞衰老、HIF-1信号通路、孕激素介导的卵母细胞成熟等途径。以上结果从活体、代谢、转录和分子层面揭示了淫羊藿对后备母猪发情的影响和作用途径,本研究发现饲喂淫羊藿能够改变后备母猪卵巢转录与代谢模式,显著提高FSH、LH和E_2水平,进而促进母猪发情,淫羊藿的主要成分能够与TP53、SRC、AKT1、CCND1、TNF、ESR1、EP300、ERBB2、JAK2、PARP1结合发挥调控作用。本研究为淫羊藿应用提高后备母猪发情利用率提供理论依据。

鱼腥草对肝癌细胞生物学行为的影响及网络药理学分析

目的探讨鱼腥草(Herba Houttuyniae)对肝癌细胞生物学行为的影响及其潜在的分子机制。方法使用不同浓度的鱼腥草提取物处理肝癌细胞SMMC-7721和SK-Hep-1,利用CCK-8检测细胞的活力,使用Transwell实验检测肝癌细胞的迁移和侵袭能力,细胞免疫荧光实验检测转移相关蛋白MMP-9的表达情况,活性氧(ROS)检测试剂盒检测细胞ROS水平,TUNEL染色检测细胞凋亡情况,Western blot检测细胞中凋亡相关蛋白表达水平。采用网络药理学筛选鱼腥草的活性成分并构建蛋白间相互作用(PPI)网络识别鱼腥草治疗肝癌的作用靶点,通过GO和KEGG通路富集分析探索与鱼腥草抗肝癌作用靶点相关的信号通路,利用qRT-PCR检测关键靶点的mRNA表达水平。结果CCK-8、Transwell、细胞免疫荧光实验结果显示鱼腥草提取物可抑制SMMC-7721和SK-Hep-1细胞的活力、迁移和侵袭能力以及转移相关蛋白MMP-9的表达(均P<0.05)。TUNEL染色、ROS检测、Western blot结果显示鱼腥草提取物可促进SMMC-7721和SK-Hep-1细胞凋亡及升高细胞中ROS水平和促凋亡蛋白BAX的表达水平(均P<0.05)。网络药理学预测鱼腥草治疗肝癌主要通过作用于TNF、AKT1、TP53等靶点,涉及癌症、TNF、PI3K-AKT等信号通路。qRT-PCR证实鱼腥草提取物可降低SMMC-7721和SK-Hep-1细胞中TNF-α、AKT1基因的mRNA表达水平,增加TP53基因的mRNA表达水平(均P<0.05)。结论鱼腥草可能通过调节TNF、AKT1、TP53等基因抑制肝癌细胞的细胞活力及迁移和侵袭能力,且促进肝癌细胞凋亡。

基于网络药理学、分子对接、实验研究探讨白屈菜治疗鼻咽癌的物质基础及潜在机制

目的运用网络药理学和分子对接技术预测白屈菜抗鼻咽癌(nasopharyngeal carcinoma,NPC)的作用靶点和机制,并通过实验验证主要活性成分对NPC细胞增殖和凋亡的影响。方法借助在线数据平台TCMSP、ETCM和BATMAN-TCM检索白屈菜的化学成分和作用靶点。通过GEO数据库检索NPC相关靶点,生信在线工具分析白屈菜与NPC的交集靶点。使用STRING构建共同靶点的PPI网络,运用Cytoscape 3.7.1获得核心靶点。通过R软件编程进行GO功能富集分析和KEGG通路富集分析。分子对接分析核心靶点与主要活性成分之间的结合情况。实验验证:(1)将5-8F细胞分为溶剂对照组、血根碱(2.5μmol·L^(-1)、5μmol·L^(-1))组、白屈菜红碱(2.5μmol·L^(-1)、5μmol·L^(-1))组、顺铂4μg·mL^(-1)组。MTT检测白屈菜主要活性成分对NPC细胞增殖的影响。(2)将5-8F细胞分为溶剂对照组、血根碱5μmol·L^(-1)组、白屈菜红碱5μmol·L^(-1)组、顺铂4μg·mL^(-1)组;Annexin-V FITC/PI双荧光染色法检测细胞凋亡;Western blot检测白屈菜主要活性成分对NPC细胞增殖、凋亡、丝裂原活化蛋白激酶(mitogen-activated protein kinase,MAPK)和磷脂酰肌醇三激酶(phosphatidylinositol 3-kinase,PI3K)/蛋白激酶B(protein kinase B,AKT)信号通路关键蛋白的影响。结果检索得到白屈菜37个主要活性成分和1419个作用靶点。检索GEO数据库共收集到7852个NPC疾病基因,白屈菜与NPC共有327个交集靶点,其中核心靶基因共10个,分别是EGFR、TP53、VEGFA、TNF、FN1、MMP9、JUN、FGF2、LYN、F2。GO分析主要涉及泛素蛋白连接酶结合、硫化物结合、整合素结合、肝素结合和糖胺聚糖结合,KEGG分析主要涉及MAPK、PI3K/AKT信号通路等,分子对接结果显示核心靶点与对应的活性成分具有良好的结合能力。实验验证显示,与溶剂对照组相比,血根碱和白屈菜红碱均明显降低NPC细胞相对增殖率(P<0.01),并提高细胞凋亡率(P<0.01),且血根碱和白屈菜红碱降低5-8F细胞中XIAP、PCNA、ERK1/2、AKT的蛋白表达水平(P<0.05或P<0.01)。结论白屈菜可通过多成分、多靶点和多通路发挥抗NPC的作用,且经实验验证,血根碱和白屈菜红碱均可抑制NPC细胞增殖并诱导凋亡,其机制可能与MAPK信号通路、PI3K/AKT信号通路有关。

基于网络药理学探究地锦草防治溃疡性结肠炎的作用机制

【目的】基于网络药理学并结合分子对接方法分析地锦草防治溃疡性结肠炎(UC)的活性成分和靶点,探究其潜在机制,进而保护动物肠道。【方法】利用TCMSP数据库获取地锦草的活性成分及对应药物靶点。通过GeneCards、DisGeNET、TTD、PharmGKB和DrugBank数据库获取疾病靶点。药物靶点和疾病靶点取交集获取潜在靶点,分别借助STRING和DAVID数据库对潜在靶点进行蛋白互作(PPI)分析及GO功能和KEGG通路富集分析。借助AutoDock 1.5.7软件进行分子对接验证,利用PyMOL软件将对接结果可视化。通过体内试验利用30只BALB/c小鼠制作葡聚糖硫酸钠(DSS)诱导的UC模型,设置对照组、模型组、地锦草低(5 mg/mL)、高(15 mg/mL)剂量组、美沙拉嗪组(52 mg/mL)。除对照组外,其余各组小鼠连续7 d自由饮用3%DSS诱导UC模型,各给药组小鼠每天灌胃1次,根据小鼠体重灌胃相应剂量的药物0.1 mL/10 g,对照组和模型组小鼠灌胃等体积无菌生理盐水,连续7 d,测其体重变化、疾病活动指数(DAI)评分和结肠长度,检测肿瘤坏死因子-α(TNF-α)、白细胞介素-6(IL6)和IL10含量。【结果】地锦草主要活性成分有山柰酚、4’-5二羟基黄酮和鞣花酸等,对应的药物靶点256个,疾病靶点4265个,潜在靶点128个,核心靶点7个。GO功能富集涉及对活性氧的反应、对氧化应激的反应、炎症反应、膜筏、小窝、激酶调节剂活性等。KEGG信号通路涉及PI3K-Akt和TNF信号通路等。分子对接结果显示,山柰酚和过氧化物酶体增殖物激活受体γ(PPARG)、4’,5-二羟基黄酮和PPARG、鞣花酸和丝氨酸/苏氨酸蛋白激酶1(AKT1)、鞣花酸和IL6具有较强结合潜力。体内试验结果表明,与对照组相比,模型组小鼠体重严重下降,严重的甚至出现水样血便,DAI评分极显著升高(P<0.01),结肠组织的黏膜层及黏膜下层出现大量炎性细胞浸润,甚至有小溃疡的出现,表明小鼠UC造模成功。与模型组相比,各给药组小鼠结肠组织黏膜层炎症细胞浸润减少,DAI评分极显著降低(P<0.01),结肠挛缩情况好转,TNF-α和IL6含量显著减少(P<0.05),IL10含量显著增加(P<0.05),抑制炎症发生。【结论】地锦草中的山柰酚、4’-5二羟基黄酮和鞣花酸等主要活性成分可能通过作用于AKT1、PPARG和IL6等核心靶点参与炎症相关信号通路及生物功能,抑制炎症,减轻肠道损伤,从而发挥防治UC的效用。

基于网络药理学和动物实验探究桑寄生对牙周炎的防治作用

目的基于网络药理学方法探究桑寄生治疗牙周炎的作用机制并通过分子对接和体内实验验证,探究桑寄生乙醇提取物对牙龈卟啉单胞菌(Porphyromonas gingivalis,P.g)生长作用的影响以及对实验性大鼠牙周炎的治疗效果。方法在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)中检索桑寄生有效作用成分,通过Swiss Target Prediction数据库预测作用靶点,经GeneCards、DisGeNET和OMIM数据库检索得到牙周炎相关靶点,通过Venny分析获得共同靶点。采用STRING数据库构建PPI网络并筛选关键基因,经DAVID数据库进行GO和KEGG富集分析,采用Cytoscape 3.10.0软件构建桑寄生“药物-成分-靶点-通路”网络,经分子对接验证药物成分与靶点的结合情况。采用倍比稀释法测定桑寄生乙醇提取物对牙龈卟啉单胞菌的最低抑菌浓度(Minimum inhibitory concentration,MIC)和最低杀菌浓度(Minimum bactericidal concentration,MBC)。将42只雄性SD大鼠随机分为空白组、溶剂组、模型组、盐酸米诺环素组、桑寄生高、中、低剂量组。丝线结扎左上第一磨牙并接种P.g构建实验性大鼠牙周炎模型,每天对各组大鼠进行相应药物干预并记录体重、探诊深度(Probing depth,PD)、龈沟出血指数(Sulcus bleeding index,SBI)。14 d后拍摄X线片确定建模成功,处死各组大鼠获取血清、颌骨及牙龈组织标本。RT-PCR检测白细胞介素6(Interleukin 6,IL-6)、肿瘤坏死因子α(Tumor necrosis factorα,TNF-α)、基质金属蛋白酶9(Matrix metalloproteinase 9,MMP-9)、Runt相关转录因子2(Runt-related transcription factor 2,RUNX2)、骨钙素(Osteocalcin,OCN)、骨保护素(Osteoprotegerin,OPG)、核因子κB受体激活配体(Receptor activator of nuclear factor-κB ligand,RANKL)mRNA表达水平。ELISA检测血清中IL-6、TNF-α、MMP-9表达水平。HE染色观察炎症浸润情况。免疫组化染色观察RUNX2、OCN蛋白表达情况。结果筛选得到4个桑寄生有效成分,545个药物靶点,4151个牙周炎治疗靶点,Venny分析得到186个桑寄生治疗牙周炎的潜在靶点。PPI网络分析得到IL-6、TNF-α、MMP9等10个核心靶点。富集分析得到的主要通路为PI3K-Akt信号通路等。分子对接结果表明桑寄生有效成分与关键靶点具有较好的结合能力。桑寄生乙醇提取物对P.g的MIC为0.5 g/L,MBC为2 g/L。桑寄生乙醇提取物对实验性大鼠牙周炎的PD、SBI有良好的改善作用,可降低血清中IL-6、TNF-α、MMP-9表达,缓解牙周组织炎症情况。可促进牙周组织中RUNX2、OCN、OPG的表达,抑制RANKL的表达,调节骨稳态,缓解牙周骨组织流失。结论桑寄生乙醇提取物对P.g具有抑制和杀灭作用,可减轻炎症反应、调节骨稳态,通过多组分、多靶点、多通路的协同作用治疗牙周炎。

基于数据挖掘和网络药理学的中医药治疗支气管哮喘的用药规律和作用机制分析

目的:探讨中医药治疗支气管哮喘的组方规律,预测核心药物潜在作用机制,以期为后续实验研究及临床应用提供依据与参考。方法:检索中国知网、万方数据库和维普数据库中采用中药复方治疗支气管哮喘的临床研究文献,对中药处方进行频次统计、关联规则分析,探究支气管哮喘的组方规律,并在此基础上以网络药理学策略,筛选高频药物核心靶点,通过基因本体功能富集分析和京都基因与基因组百科全书通路富集分析,研究药物对疾病的潜在作用靶点和通路,以期阐明其作用机制。结果:共纳入448项研究,包含459首处方、210味中药;其中,化痰止咳平喘药(占28.70%,1 413次/4 924次)和补虚药(占21.83%,1 075次/4 924次)应用较多,以温性(占49.31%,2 483次/5 035次)、辛味(占29.75%,2 295次/7 713次)为主,归经以入肺经(占29.08%,3 608次/12 408次)最多;关联分析得到高频药对与高频角药各10组。麻黄为治疗支气管哮喘的核心药物,其与支气管哮喘共有147个交集靶点,主要有白天竺葵苷元、草本菌素和无色矢车菊素等23种活性成分,核心靶点为热休克蛋白90AA1、肿瘤蛋白p53和JUN等,主要通路为癌症信号通路、脂质与动脉粥样硬化通路、糖尿病并发晚期糖基化终末产物(AGE)-AGE受体信号通路等。结论:中医药治疗支气管哮喘注重清肺化痰、止咳平喘及健脾益肾,兼顾解表与清热类药物的应用;高频药物麻黄的潜在靶点和作用机制主要参与药物的反应、基因表达的正向调节及凋亡过程的负调控等生物过程,发挥治疗作用。

基于数据挖掘探究溪黄草药用成分及潜在抗癌的作用机制

目的探讨溪黄草的药效成分及其潜在的抗肿瘤作用机制。方法通过文献检索、BATMAN-TCM数据库及CancerHSP数据库收集溪黄草的化学成分,利用化源网查询CAS号,通过TCMSP数据库和SwissADME在线平台筛选有效成分;运用SwissTargetPrediction在线平台预测各有效成分的靶点,通过Metascape数据库对靶点进行GO和KEGG富集分析,通过在线作图平台微生信将结果可视化。通过STRING平台获取蛋白互相作用网络并筛选关键靶点,通过NIMNT数据库进行关键靶标的适应证预测,采用网络绘图软件Cytocape3.9.1构建“药物-核心靶标-疾病”互作网络。结果共筛选到5,6,3′,4′-四羟基-7-甲氧基黄酮、铁锈醇、山柰酚和杜鹃黄素等10个有效成分,这些成分作用于212个靶点,其中SRC、PIK3R1、PIK3CA、AKT1、ESR1、CDK1和EGFR等为核心靶点。GO富集分析得到生物过程条目5310条,细胞组成条目524条,分子功能条目981条;KEGG通路富集筛选获得260条信号通路,参与癌症通路、磷酸酶D信号通路和cGMP-PKGX信号通路等多条通路。结论溪黄草抗癌活性是多成分、多靶点、多通路的作用结果,为临床合理使用中草药辅助治疗癌症提供了一定的科学理论依据。