Network pharmacology-based exploration of molecular mechanisms underlying therapeutic effects of Jianpi Huatan Quyu recipe on chronic heart failure with spleen Qi deficiency syndrome

BACKGROUND Chronic heart failure is a complex clinical syndrome.The Chinese herbal compound preparation Jianpi Huatan Quyu recipe has been used to treat chronic heart failure;however,the underlying molecular mechanism is still not clear.AIM To identify the effective active ingredients of Jianpi Huatan Quyu recipe and explore its molecular mechanism in the treatment of chronic heart failure.METHODS The effective active ingredients of eight herbs composing Jianpi Huatan Quyu recipe were identified using the Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform.The target genes of chronic heart failure were searched in the Genecards database.The target proteins of active ingredients were mapped to chronic heart failure target genes to obtain the common drugdisease targets,which were then used to construct a key chemical componenttarget network using Cytoscape 3.7.2 software.The protein-protein interaction network was constructed using the String database.Gene Ontology and Kyoto Encyclopedia of Genes and Genomes enrichment analyses were performed through the Metascape database.Finally,our previously published relevant articles were searched to verify the results obtained via network pharmacology.RESULTS A total of 227 effective active ingredients for Jianpi Huatan Quyu recipe were identified,of which quercetin,kaempferol,7-methoxy-2-methyl isoflavone,formononetin,and isorhamnetin may be key active ingredients and involved in the therapeutic effects of TCM by acting on STAT3,MAPK3,AKT1,JUN,MAPK1,TP53,TNF,HSP90AA1,p65,MAPK8,MAPK14,IL6,EGFR,EDN1,FOS,and other proteins.The pathways identified by KEGG enrichment analysis include pathways in cancer,IL-17 signaling pathway,PI3K-Akt signaling pathway,HIF-1 signaling pathway,calcium signaling pathway,cAMP signaling pathway,NF-kappaB signaling pathway,AMPK signaling pathway,etc.Previous studies on Jianpi Huatan Quyu recipe suggested that this Chinese compound preparation can regulate the TNF-α,IL-6,MAPK,cAMP,and AMPK pathways to affect the mitochondrial structure of myocardial cells,oxidative stress,and energy metabolism,thus achieving the therapeutic effects on chronic heart failure.CONCLUSION The Chinese medicine compound preparation Jianpi Huatan Quyu recipe exerts therapeutic effects on chronic heart failure possibly by influencing the mitochondrial structure of cardiomyocytes,oxidative stress,energy metabolism,and other processes.Future studies are warranted to investigate the role of the IL-17 signaling pathway,PI3K-Akt signaling pathway,HIF-1 signaling pathway,and other pathways in mediating the therapeutic effects of Jianpi Huatan Quyu recipe on chronic heart failure.

Mechanism of Jianpi Qingchang Huashi Recipe in treating ulcerative colitis:A study based on network pharmacology and molecular docking

BACKGROUND Ulcerative colitis(UC)is a refractory intestinal disease with alternating onset and remission and a long disease course,which seriously affects the health and quality of life of patients.The goal of treatment is to control clinical symptoms,induce and maintain remission,promote mucosal healing,and reduce recurrence.Clinical trials have shown unsatisfactory clinical response rates.As a supplementary alternative medicine,traditional Chinese medicine has a rich history and has shown good results in the treatment of UC.Because of the quality of herbal medicine and other factors,the curative effect of traditional Chinese medicine is not stable enough.The mechanism underlying the effect of Jianpi Qingchang Huashi Recipe(JPQCHSR)on inducing UC mucosal healing is not clear.AIM To investigate the potential mechanism of JPQCHSR for the treatment of UC based on network pharmacology and molecular docking.METHODS Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform was used to extract the active components and action targets of JPQCHSR,and the target names were standardized and corrected through UniProt database.The related targets of UC were obtained through GeneCards database,and the intersection targets of drugs and diseases were screened by jvenn online analysis tool.The visual regulatory network of"Traditional Chinese medicine-active components-target-disease"was constructed using Cytoscape software,the protein interaction network was constructed using STRING database,and enrichment analysis of gene ontology and Kyoto Encyclopedia of Genes and Genomes pathways was conducted through R software.At last,the active components were docked with the core target through SYBYL-X 2.1.1 software.RESULTS Through database analysis,a total of 181 active components,302 targets and 205 therapeutic targets were obtained for JPQCHSR.The key compounds include quercetin,luteolin,kaempferol,etc.The core targets involved STAT3,AKT1,TP53,MAPK1,MAPK3,JUN,TNF,etc.A total of 2861 items were obtained by GO enrichment analysis,and 171 items were obtained by KEGG(Kyoto Encyclopedia of Genes and Genomes)pathway enrichment analysis.The results of molecular docking showed that the key active components in JPQCHSR had certain affinity with the core target.CONCLUSION The treatment of UC with JPQCHSR is a complex process of multi-component,multi-target and multi-pathway regulation.The mechanism of this Recipe in the treatment of UC can be predicted through network pharmacology and molecular docking,so as to provide theoretical reference for it to better play its therapeutic role.

Analyzing the effective compounds, potential targets and diseases of Jianpi Jiedu recipe based on network pharmacology and function validation of cytobiology

Objective: To analyze the active compounds, potential drug targets and therapy diseases of Jianpi Jiedu Recipe (JPJDR) based on network pharmacology and bioinformatics technology, and verify the biological function of some active compounds by cytology experiments. Methods: The online databases including TCMSP, TCMID, Cancer HSP, TCM-PTD, TCM Database@Taiwan and DrugBank were applied to screen the active compounds and the potential drug targets of JPJDR. Cytoscape 3.3 software was executed to construct the network between active compounds and drug targets. DAVID database was used to probe the effective diseases and analyze the involved KEGG pathways according to the predicted targets corresponding to JPJDR. Results: According to the rules of oral bioavailability (OB)>30% and drug-likeness (DL)>0.18, 58 of 513 effective compounds in JPJDR were screened out, as well as the corresponding 437 potential drug targets. By the analysis of DAVID database, all these key targets were associated closely with the occurrence and development of metabolic disorders and cancers, and all the targets were closely correlated with the pathways in cancer. Further analysis demonstrated that, there were a lot of effective compounds in JPJDR, such as Quercetin, Formononetin, Stigmasterol, Diosgenin,β-sitsterol, Oxymatrine, Kaempferol, Isorhamnetin and Ampelopsis. The results of cell proliferation experiments further showed that, among the selected nine key traditional Chinese medicine compounds, only Ampelopsis can dose-dependently inhibit the proliferation of colorectal cancer cells. Conclusions: Through network pharmacology analysis, we found that JPJDR contains many effective compounds which may directly target to the cancer-related proteins. 9 compounds were the major active compounds with high degrees of targets. Among the 9 screened compounds, Ampelopsis was validated for its inhibitory effect on the proliferation of colorectal cancer cells using CCK-8 assay. Network pharmacology is an effective approach to explore the functional mechanism of formula.

Wenjing Zhitong recipe exhibits potential anti-primary dysmenorrhea properties by inhibiting COX2 and PKC signaling pathway in rats induced by estradiol benzoate and oxytocin

Objective:To explore the effect and mechanism of action of the Wenjing Zhitong recipe(WZR)in primary dysmenorrhea(PD)treatment.Methods:Uterine contractions were induced by estradiol benzoate and oxytocin in a PD model and WZR was administrated.The rate of change in uterine contractility and the writhing test were used to evaluate the effects of WZR.The serum levels of prostaglandin F_(2a)(PGF_(2a))and prostaglandin E_2(PGE_2),and the activity of cyclooxygenase-2(COX2)were detected by enzyme-linked immunosorbent assay(ELISA).The changes in phosphor-phospholipase C(pPLC/PLC),phosphor-protein kinase C(pPKC/PKC),and connexin 43(CX43)expression were detected using immunohistochemistry and western blot.Results:WZR significantly reduced the rate of change in uterine contractility and writhing times in the PD model.WZR treatment inhibited the enzymatic activity of COX2 and reduced the levels of PGF_(2a),PGF_(2a)/PGE2and COX2 in the PD model.WZR also significantly reduced the expression of pPLC/PLC,pPKC/PKC and CX43.Targeting the inhibition of COX2 activity,caffeic acid and 1-acetyl-β-carboline were validated as the active ingredients in WZR responsible for reducing uterine contractions.Conclusion:WZR attenuated PD by inhibiting COX2 activity,downregulating PGF_(2a)/PGE_2 expression,and inhibiting the PKC signaling pathway.

Mechanism of Xingshen Zhiyi prescription in the treatment of enuresis based on network pharmacology

Objective:To explore the mechanism of Xingshen Zhiyi Recipe(XSZYF)in the treatment of Nocturnal Enuresis(NE)based on network pharmacology.Methods:TCMSP,DrugBank databases,PubMed and CNKI were used to obtain the active ingredients and corresponding targets of XSZYF.NE targets were obtained from GeneCard and OMIM databases.Cytoscape software was used to construct a drug-disease-target network model.The analysis was performed.The protein interaction network(PPI)was constructed using the STRING database.The gene ontology functional annotation(GO)and the Tokyo Genomic Encyclopedia(KEGG)pathway enrichment analysis were performed on key targets using the DAVID online tool.Surflex docking software was used for the analysis.Docking of key active ingredients and key targets to verify the results of network analysis.Results:199 gene targets of XSZYF were obtained,and 2486 gene targets of NE.Network analysis results showed that the key targets of XSZYF for treating NE include CHRM3,CHRM2,ADRB3,etc.Involved in regulating neuroactive ligand-receptor interactions,calcium signaling pathways,etc.Conclusion:This study revealed the material basis and action mechanism of XSZYF in treating NE from the perspective of network pharmacology.

Effective ingredients, potential targets and mechanism in cancers treatment of Bushen Jiedu Sanjie recipe

Objective: To analyze the active compounds, potential drug targets and corresponding therapy cancer of Bushen Jiedu Sanjie Recipe (BSJDSJR) based on network pharmacology and bioinformatics technology. Methods: The network databases including Cancer HSP, TCMSP, TCMID, TCM-PTD, TCM Database@Taiwan and DrugBank were applied to screen the active compounds, potential drug targets and corresponding cancers of BSJDSJR. Cytoscape3.3 software was used to construct the network between active compounds of Chinese medicine and the corresponding targets. Then, the enrichment of biological processes and KEGG pathways of BSJDSJR were analyzed using DAVID database. Results: According to Oral bioavailability (OB)≥30% and drug like index (DL)≥0.18, 129 active compounds in BSJDSJR were screened out and they targeted to 301 proteins. These targets were closely associated with the occurrence of various cancers, such as bladder cancer, pancreatic cancer, non-small cell lung cancer and colorectal cancer. Further investigation showed that, there were lots of active compounds in BSJDSJR are closely connected with the COX-2/β-catenin signaling pathway, STAT3 signaling pathway and MAPK/ERK signaling pathway. Conclusions: Based on the network pharmacology, the study disclosed the active chemical compounds, potential targets and possible action cancers of BSJDSJR.

益气血补肝肾方对胚胎反复种植失败患者冻融胚胎移植周期妊娠结局的影响

目的探讨益气血补肝肾方对胚胎反复种植失败(RIF)患者冻融胚胎移植(FET)周期妊娠结局的影响。方法回顾性分析2017年1月至2023年6月在广州市第一人民医院生殖健康与不孕症专科行FET的120例RIF患者临床资料。患者月经周期规律,年龄25~35岁,按照FET前是否接受益气血补肝肾方治疗分为对照组和研究组,各60例;另选取同时期57例既往无胚胎移植史的FET患者作为正常组。3组均采用激素替代(HRT)方案准备内膜。研究组在准备内膜前先给予益气血补肝肾方3个月经周期(经后增殖方:煎成20 ml,制成颗粒剂10 g,1包/d,早晚分2次服用,月经干净后开始服用至排卵期;促黄体方:煎成20 ml,制成颗粒剂10 g,1包/d,早晚分2次服用,自排卵后开始服用至月经第1~3天停药)。比较3组患者内膜转化日子宫内膜厚度、血流搏动指数(PI)、动脉阻力指数(RI)、胚胎着床率和临床妊娠率。统计学方法采用单因素方差分析和χ^(2)检验。结果正常组和研究组子宫内膜厚度均高于对照组,PI、RI均低于对照组,3组子宫内膜厚度、PI、RI比较[(10.33±2.27)mm比(10.24±1.81)mm比(9.03±1.41)mm、(1.86±0.52)比(1.96±0.21)比(1.97±0.20)、(0.56±0.10)比(0.58±0.90)比(0.62±0.09)],差异均有统计学意义(F=8.40、6.08、6.89,均P<0.05)。正常组和研究组胚胎着床率、临床妊娠率均高于对照组,3组比较差异均有统计学意义[43.9%(43/98)比38.8%(40/103)比25.2%(26/103)、57.9%(33/57)比55.0%(33/60)比35.0%(21/60);χ^(2)=8.18、7.37,均P<0.05]。结论益气血补肝肾方可通过降低子宫内膜血流阻力指数和搏动指数,增加子宫内膜厚度,提高胚胎着床率和临床妊娠率,改善RIF患者FET周期妊娠结局。

基于网络药理学及分子对接探讨润肝养心方治疗心脏神经症的分子机制

目的:基于网络药理学和分子对接方法探讨润肝养心方治疗心脏神经症(CN)的物质基础和作用机制。方法:通过中药系统药理学数据库与分析平台(TCMSP)、有机小分子生物活性数据库(PubChem)、Swiss Target Prediction数据库检索并筛选出润肝养心方(麦冬、酸枣仁、柏子仁、炒白芍、当归、五味子、茯苓、炙甘草)的药物活性成分及作用靶点,利用基因名片数据库(GeneCard)、在线人类孟德尔遗传数据库(OMIM)、DisGeNET数据库筛选CN疾病靶点,将药物和疾病靶点两者取交集后获得的共有靶点导入STRING数据库获得蛋白互作关系,应用Cytoscape 3.8.2软件构建蛋白质相互作用(PPI)网络;采用R语言ClusterProfiler程序包(3.18.0)对关键靶点进行基因本体(GO)和京都基因和基因组百科全书(KEGG)富集分析,并利用Cytoscape 3.8.2构建关键靶点-功能-通路图及关键靶点中药药理调控网络;采用AutoDock Tools 1.5.6进行分子对接研究。结果:润肝养心方筛选得到活性成分146个、作用靶点245个、CN及润肝养心方共有靶点如白细胞介素(IL)-1β、IL-6、肿瘤坏死因子(TNF)等38个。KEGG富集分析关键靶点主要被富集在神经活性配体-受体相互作用、晚期糖基化终末产物(AGE)-糖基化终末产物受体(RAGE)信号通路、脂质与动脉粥样硬化等信号通路,涉及对异源刺激的反应、管径调节、血管直径维持等生物过程。分子对接验证显示关键靶点与槲皮素、芍药苷、山柰酚的结合活性较好。结论:润肝养心方中主要活性成分可能通过调节IL-1β、IL-6、TNF等关键靶点基因及神经活性配体-受体相互作用、AGE-RAGE信号通路、脂质与动脉粥样硬化等多通路以发挥抗炎、调节神经递质水平等作用治疗CN。

基于指纹图谱和网络药理学对经典名方五味消毒饮质量标志物的预测分析

目的 基于HPLC指纹图谱和网络药理学对五味消毒饮中的质量标志物(Q-marker)进行预测。方法 建立15批五味消毒饮的指纹图谱,并对其进行峰指认和峰归属,利用中药色谱指纹图谱相似度评价系统软件(2012年版)进行分析;通过网络药理学对核心靶点及通路进行筛选,构建“成分-靶点-通路”网络图;以Q-marker“五原则”为核心预测五味消毒饮的质量标志物。结果 15批五味消毒饮指纹图谱相似度大于0.95,标定13个共有峰,对其中11个特征峰进行指认,分别为新绿原酸、绿原酸、秦皮乙素、隐绿原酸、马钱子苷、断氧化马钱子苷、菊苣酸、异绿原酸B、异绿原酸A、异绿原酸C、蒙花苷;通过网络药理学分析,推测绿原酸、秦皮乙素、断氧化马钱子苷、菊苣酸、蒙花苷5个成分可能为五味消毒饮的潜在Q-marker,其可能通过调节肿瘤抑制因子p53/癌蛋白Mdm2、酪氨酸蛋白激酶SRC等多个基因以及胰腺癌、前列腺癌等多条癌症疾病通路来发挥抗肿瘤、抗癌功效;同时还对五味消毒饮可用于治疗新型冠状病毒肺炎进行预测分析。结论 建立的五味消毒饮HPLC指纹图谱分析方法灵敏、快速且稳定;结合网络药理学筛选出具有溯源性和可预测性,且与五味消毒饮临床应用相关的5个化合物作为其潜在Q-marker,为今后五味消毒饮的进一步研究提供参考。

基于网络药理学、分子对接及实验研究探讨活血解毒降糖方治疗糖尿病心肌病的作用机制

目的:运用网络药理学、分子对接和动物实验探讨活血解毒降糖方治疗糖尿病心肌病(DCM)的主要化学成分及其作用机制。方法:利用网络药理学工具,确定治疗DCM的活血解毒方有效成分、核心靶点和信号通路。建立DCM大鼠模型以验证网络药理学预测的主要靶点及治疗效果。结果:筛选出活血解毒降糖方158个活性成分,涉及318个DCM靶点及204条相关信号通路。网络分析表明,活血解毒降糖方可能通过磷酸酶基因(PTEN)、雷帕霉素靶蛋白(mTOR)、肿瘤蛋白P53(TP53)、信号转导与转录激活因子3(STAT3)、血管内皮细胞生长因子(VEGFA)等关键靶点治疗DCM。分子对接显示活血解毒降糖方对活性成分槲皮素、γ-氨基丁酸、腺苷、木犀草素、山柰酚成分与PTEN、mTOR、TP53、STAT3、VEGFA等关键靶点结合性较好,其中,木犀草素与mTOR的Vina得分最低。动物实验结果表明,活血解毒降糖方可以减轻心肌纤维化,降低C反应蛋白(CRP)、肿瘤坏死因子-α(TNF-α)、白细胞介素(IL)-6水平,并抑制mTOR蛋白的表达,从而发挥治疗DCM作用。结论:活血解毒降糖方通过多成分、多靶点的作用改善DCM,为进一步探究活血解毒降糖方治疗DCM提供了研究思路。

溶髓方干预终板软骨细胞凋亡作用机制研究:基于网络药理学与分子对接技术

目的:应用网络药理学和分子对接技术,探讨溶髓方干预终板软骨细胞凋亡(CA)的潜在作用机制,为治疗颈椎间盘退行性病变(CIDD)寻找新思路。方法:通过中药系统药理学分析平台(TCMSP)和中医药百科全书(ETCM),获取溶髓方中13种药物的活性成分和预测分子潜在作用靶点。检索Gennecards和OMIM数据库,筛选出与CA进程相关的靶基因,并与药物靶点取交集。应用Cytoscasp 3.7.2软件构建“中药-活性成分-靶点”网络并借助CytoNCA绘制核心靶点拓扑网络进行拓扑分析,应用STRING数据库构建蛋白互作网络并筛选出核心靶点,使用R4.2.6软件对差异基因进行GO功能富集分析、KEGG通路富集分析。通过AutoDock4和Python3.2对获得的核心成分和核心靶点进行分子对接。结果:(1)获得溶髓方治疗CA的154个活性成分,主要为槲皮素、γ-谷甾醇、山柰酚、黄芩素、桑辛素C等,通过拓扑网络筛选出关键候选基因肿瘤蛋白P53(TP53)、丝裂原活化蛋白激酶3(MAPK3)、Jun原癌基因(JunProto-Oncogene,JUN)等。KEGG富集分析显示,与CA治疗相关的信号通路主要为AGE-RAGE信号通路、IL-17信号通路、TNF信号通路、NF-κB信号通路;(2)分子对接结果显示,溶髓方中核心有效成分与CA核心靶点均能通过氢键结合,且分子对接结合能均≤-5.0 kcal/mol,显现出良好的结合力。结论:溶髓方干预CA进程具有多成分、多靶点、多途径等特征,作用机制可能是溶髓方中的槲皮素、γ-谷甾醇、山柰酚、黄芩素、桑辛素C等活性成分作用于TP53、MAPK3、JUN等靶点,通过调控AGE-RAGE信号通路、IL-17信号通路、TNF信号通路、NF-κB信号通路等信号通路发挥干预CA进程的作用。

Yiqixue Buganshen Recipe(益气血补肝肾方) Regulates the Expression of Integrin ανβ 3 in the Endometrium of Controlled Ovarian Hyperstimulation Mice

Objective： To observe the effect of Yiqixue Buganshen recipe （益气血补肝肾方, YBR） on the expression of integrin e~ v 13 3 in the endometrium of controlled ovarian hyperstimulation mice. Methods： A total of 180 mice were divided into three groups： model group, treatment group and control group. The treatment and model groups were intraperitoneally injected with gonadotropin-releasing hormone analogue for 7 days; pregnant mare serum gonadotropin was also injected on the 7th day. After 48 h, human chorionic gonadotropin was injected. The control group was injected with an equal volume of saline at the same time. From the start of the experiment, the treatment group was intragastrically administered Jinghouzengzhi Recipe （经后增殖方） and Cuhuangti Recipe （促黄体方）. The model group and the control group were intragastrically administered an equal volume of saline. Real-time reverse transcription polymerase chain reaction and Western blotting were used to detect the mRNA and protein expression of integrin α υ β 3 in mouse endometrium. Results： Integrin α υ β 3 was expressed in mouse endometrium in all groups. Integrin α υ β 3 expression increased gradually along with pregnancy, progressing from pregnant day （Pd） 1. Integrin α υ β 3 expression significantly increased on Pd 4, then began to decrease on Pd 6. Integrin α υ β 3 expression in the treatment group was higher than in the model group, and the difference was statistically significant （P〈0.05）. The difference between the treatment group and the control group was not statistically significant （P〉0.05）. Conclusion： YBR improves endometrial receptivity, and may play an important role in embryonic implantation.

Mechanism of Huashi Xingyu Qingre recipe(化湿行淤清热方)in treating oral lichen planus based on network pharmacology and clinical trial verification

OBJECTIVE:To identify the main active components and targets of Huashi Xingyu Qingre recipe(化湿行淤清热方,HXQR)and to investigate its mechanism in the treatment of oral lichen planus(OLP).METHODS:The Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform was searched to identify the active ingredients and corresponding targets of HXQR.Disease genes were obtained from the Gene Cards database,and a“drugdisease regulatory network”was constructed using Cytoscape software and PERL programming language.The STRING database was used to build a protein-protein interaction network.Gene ontology(GO)and Kyoto Encyclopedia of Genes and Genomes(KEGG)terms were analyzed using R software with a Bioconducter plugin.Finally,the results and the efficacy of HXQR in treating OLP were validated in a clinical trial that included enzyme-linked immunosorbent assay(ELISA)testing and observations of the post-treatment changes in clinical symptoms.RESULTS:HXQR contained 167 active components and 261 targets,with 391 disease targets.The intersection of these two categories in a Venn diagram revealed 57 drugdisease common targets.A compound-target network was constructed and revealed that the six key pharmaceutical ingredients of HXQR were quercetin,luteolin,wogonin,kaempferol,beta-carotene,and baicalein.The protein-protein interaction network mainly involved core proteins such as ALB,interleukin-6,and AKT1.Drug-disease common targets were enriched in 1628 GO terms and 117 KEGG terms,mainly involving inflammatory responses,viral infections,and tumorrelated pathways.ELISA testing indicated that HXQR inhibited the tumor necrosis factor(TNF)signaling pathway by reducing the expression of interleukin-6,matrix metalloproteinase-9,and intercellular adhesion molecule-1.The clinical symptoms of the patients with OLP were significantly improved after 8 weeks of treatment with HXQR.CONCLUSION:HXQR treats OLP by regulating the TNF signaling pathway,resulting in a marked treatment effect with few adverse effects.

基于血清药物化学、网络药理学及细胞实验探究壮骨健膝方治疗膝骨关节炎的药效物质

目的探究并验证壮骨健膝方(骨碎补、杜仲、秦艽、独活等组成)治疗膝骨关节炎(knee osteoarthritis,KOA)的药效物质。方法基于壮骨健膝方药物成分自建数据库,采用超高效液相串联三重四级杆质谱技术(ultra performance liquid chromatography tandem mass spectrometry,UPLC-MS/MS)定性定量检测壮骨健膝方入血成分;通过TCMSP、SwissTargetPrediction、SEA和Batman-TCM数据库筛选入血成分靶点,以“knee osteoarthritis”为关键词在GeneCards、OMIM、Drugbank数据库搜索膝骨关节炎相关靶点,将二者取交集后,通过Cytoscape软件构建入血成分-膝骨关节炎作用靶点网络,STRING平台构建蛋白互作网络,寻找核心成分与靶点;通过DAVID数据库对靶点进行基因本体(GO)功能富集和京都基因与基因组百科全书(KEGG)通路富集分析;通过细胞实验,以核心成分在血清中的浓度干预炎症滑膜巨噬细胞、退变软骨细胞模型,ELISA法检测细胞上清中膝骨关节炎核心靶点指标的含量。结果在壮骨健膝方含药血清中定性定量检测出6种原型入血成分,各成分及浓度分别为龙胆苦苷(1002.02±51.60)ng·mL^(-1),佛手柑内酯(448.50±13.35)ng·mL^(-1),阿魏酸(225.60±9.61)ng·mL^(-1),獐牙菜苦苷(68.49±4.62)ng·mL^(-1),柚皮苷(2.34±0.09)ng·mL^(-1),柚皮素(1.69±0.07)ng·mL^(-1);6种入血成分靶点406个,膝骨关节炎疾病靶点2492个,交集靶点155个,度值排前3位的核心成分分别是阿魏酸、柚皮素、龙胆苦苷,核心靶点包括肿瘤坏死因子(tumor necrosis factor,TNF)、白细胞介素1β(interleukin-1 beta,IL-1β)、基质金属蛋白酶(matrix metalloproteinases,MMPs)、丝裂原活化蛋白激酶3(mitogen-activated protein kinase 3,MAPK3)等;功能富集分析表明,壮骨健膝方通过多途径发挥其在膝骨关节炎中的药理作用,主要涉及炎症反应、凋亡过程、脂多糖的反应等;细胞实验结果显示,与模型组比较,阿魏酸组、柚皮素组、龙胆苦苷组均可明显抑制炎症滑膜巨噬细胞上清中IL-1β、TNF-α表达(P<0.05);三者均可明显抑制退变软骨细胞上清中MMP-3、MMP-13表达(P<0.05)。结论通过血清药物化学、网络药理学及细胞实验三方面的相互印证,揭示了阿魏酸、柚皮素、龙胆苦苷可能是壮骨健膝方治疗膝骨关节炎的核心药效物质,为该方深入开发和应用提供了客观依据。

基于网络药理学-分子对接探讨益气养阴通络方抗类风湿关节炎的作用机制

[目的]采用网络药理学与分子对接探究益气养阴通络方干预类风湿关节炎(RA)的作用机制。[方法]通过中药系统药理学数据库及分析平台(TCMSP)、本草组鉴(HERB)和文献查阅获得复方活性成分与靶点,采用Genecards、在线人类孟德尔遗传数据库(OMIM)等数据库获得疾病靶点,然后采用数据库计算出益气养阴通络复方成分与疾病的交集靶点。对交集靶点施行功能和通路富集分析,选取益气养阴通络方核心药物靶点与关键化合物施行分子对接。[结果]网络药理学共筛选出益气养阴通络方的83种活性成分和779个潜在靶点,RA相关靶点5393个,交集靶点465个,其中交集核心靶点9个(分别为SRC、STAT3、MAPK3、MAPK1、HSP90AA1、PIK3R1、GRB2、AKT1、PIK3CA)。富集得出PI3K-AKT、癌症相关及神经活性配体-受体相互作用等通路,其中PI3K-AKT信号通路可能是复方干预RA的关键通路之一。对接计算表明,益气养阴通络方的11种主要活性成分与关键靶点SRC、STAT3、PIK3R1、AKT1均有较稳定的结合,其中与AKT的结合可能最大。[结论]益气养阴通络方通过多成分-靶点-通路的方式干预RA,PI3K-AKT、癌症相关及神经活性配体-受体相互作用通路均为可能的作用通路。

基于网络药理学探讨参麻益智方配伍规律及其治疗血管性痴呆的合理性分析

目的:基于网络药理学方法分析阐释参麻益智方的配伍规律及其治疗血管性痴呆(VaD)的合理性。方法:对参麻益智方中4味中药人参、鬼箭羽、天麻、川芎、全方及不同配伍的预测靶点进行京都基因和基因组百科全书(KEGG)信号通路富集分析,构建“成分-靶点”网络等,发现每味中药及全方可能参与的信号通路。通过STRING网站构建参麻益智方靶点和VaD发病共同靶点的蛋白-蛋白互作(PPI)网络,并进行KEGG信号通路富集分析,发现参麻益智方治疗VaD的关键干预靶点及可能的作用机制。结果:参麻益智方的预测靶点共291个。KEGG信号通路富集分析发现每味中药及全方都参与代谢及血管功能调节。人参及天麻的共同作用靶点93个,人参及川芎的共同作用靶点91个,天麻和川芎可协助君药人参改善机体血管功能。参麻益智方靶点和VaD发病的共同靶点主要包括白细胞介素6、肿瘤坏死因子、载脂蛋白E、半胱氨酸蛋白酶3及脑源性神经营养因子等。KEGG信号通路富集分析显示参麻益智方治疗VaD的主要调控部位在血管,与改善脑内血管炎症有关。结论:参麻益智方配伍严谨、合理,其治疗VaD可能与改善脑血管内皮细胞炎症有关。

张忠德顽咳方治疗肺气虚寒证咳嗽(慢性支气管炎)的网络药理学机制研究

目的探讨张忠德教授治疗肺气虚寒证咳嗽(慢性支气管炎)验方顽咳方(化橘红、细辛、半夏、前胡、紫苑等)的药理机制。方法从中医药系统药理数据库和分析平台(TCMSP)获取顽咳方的有效成分及作用靶点,运用MalaCards和比较毒物基因组学数据库(CTD)获取慢性支气管炎疾病基因,使用WebGestalt在线基因集分析工具进行基因本位论(GO)和京都基因与基因组百科全书(KEGG)信号通路富集分析,运用STRING在线工具构建蛋白互相作用网络,使用Cytoscape软件进行网络可视化,运用MCODE和CytoHubba软件筛选网络核心靶点,运用CB-DOCK 2对关键靶点和成分进行分子对接。结果顽咳方靶点与慢性支气管炎疾病的共有靶点共81个,构建了“药物-成分-共有靶点”网络。共有靶点显著富集的GO条目涉及细胞运动、氧代谢、脂多糖、皮质醇的代谢反应和调节、细胞因子的活性及受体结合等相关生物过程,富集的KEGG信号通路有肿瘤坏死因子(TNF)、Th17/IL-17等炎症和免疫相关通路、松弛素、Toll样及NOD样受体等信号转导通路。共有靶点网络前2位的核心靶点为TNF和IL-6,核心子网络共4个,均与TNF、Th17/IL-17等炎症和免疫途径、松弛素信号通路等密切相关。结论顽咳方治疗慢性支气管炎的机制可能主要通过调节TNF、Th17/IL-17等炎症和免疫相关通路、松弛素、Toll样及NOD样受体信号通路发挥治疗作用。

基于网络药理学分析益气活血化痰方抗动脉粥样硬化的作用机制

目的:采用网络药理学探讨益气活血化痰方治疗动脉粥样硬化(AS)的作用机制。方法:利用中药系统药理学数据库分析平台(TCMSP)及文献检索,筛选出黄芪、丹参、全瓜蒌、黄连及水蛭的有效成分,借助有机小分子生物活性数据库(PubChem)、基因名片数据库(GeneCards)等获得本方及疾病靶点,绘制韦恩图后获得共同靶点,利用SRTING数据库及Cytoscape 3.9.1软件构建药物-疾病靶点蛋白质互作(PPI)网络图,筛选出关键靶点。使用David数据库对关键靶点进行基因本体(GO)功能富集分析及京都基因与基因组百科全书(KEGG)通路富集分析,以Cytoscape 3.9.1行可视化分析。结果:获取益气活血化痰方与AS的共同靶点265个、关键靶点36个,其中,蛋白激酶B1(AKT1)、禽肉瘤病毒17假定转化基因(JUN)、丝裂原活化蛋白激酶1(MAPK1)、p38丝裂原活化蛋白激酶14(MAPK14)、丝裂原活化蛋白激酶3(MAPK3)、核因子Kappa Bp65(RELA)、肿瘤坏死因子(TNF)等在益气活血化痰方抗AS过程中尤为重要。获得122条相关通路,主要包括癌症的通路、脂质和AS通路、糖尿病并发症中的晚期糖基化终产物及其受体(AGE-RAGE)信号通路在糖尿病并发症中的应用、卡波西肉瘤相关性疱疹病毒感染、蛋白质聚糖在癌症中的应用等。结论:益气活血化痰方通过多靶点、多通路发挥抗AS作用,可为益气活血化痰方治疗AS的临床应用提供理论依据。

基于网络药理学和生物信息学探讨扶正养心方治疗心力衰竭合并糖尿病的机制

目的:运用网络药理学与生物信息学方法探讨扶正养心方治疗心力衰竭合并糖尿病的作用成分和机制。方法:基于GEO数据库筛选心力衰竭合并糖尿病与心力衰竭的差异基因;BATMAN-TCM、ETCM数据库检索扶正养心方的药物活性成分和作用靶点,将获得的疾病基因和药物成分靶点取交集绘制Venn图及中药复方调控网络。对交集基因进行蛋白-蛋白互作(PPI)分析、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。应用分子对接技术验证扶正养心方和核心靶点之间的关系。结果:共获得差异表达基因108个;扶正养心方中含有活性成分403个、作用靶点2 346个;交集基因15个。缺口受体蛋白1(NOTCH1)为PPI核心网络中的关键蛋白。GO富集分析包括565个生物过程、13种细胞组分、4个分子功能。KEGG通路富集分析主要包括RAS相关蛋白1(RAP1)信号通路、磷脂酰肌醇3-激酶-蛋白激酶B信号通路、磷脂酶D信号通路、肌动蛋白细胞骨架调节、丝裂原活化蛋白激酶信号通路等18条信号通路,血小板衍生生长因子受体β(PDGFRB)、凝血因子Ⅱ凝血酶受体(F2R)和KIT原癌基因酪氨酸蛋白激酶(KIT)是通路富集最多的基因。分子对接结果显示,扶正养心方的核心成分与NOTCH1、PDGFRB、F2R和KIT具有较好的结合力。结论:扶正养心方干预心力衰竭合并糖尿病具有多成分、多靶点、多通路的特点。

基于网络药理学和分子对接探讨宣肺润燥解毒方治疗新冠肺炎的作用机制

目的运用网络药理学和分子对接技术,探究宣肺润燥解毒方治疗新冠肺炎的分子机制。方法利用中药系统药理学数据库与分析平台(TCMSP)、TCMID数据库筛选宣肺润燥解毒方活性成分,利用有关数据库筛选新冠肺炎相关靶点。以Cytoscape软件构建“药物-化合物-靶点”网络,并用分子对接验证有效成分与靶点亲和力。结果筛选出宣肺润燥解毒方有效成分3019个,潜在靶点525个。分子对接结果显示活性成分槲皮素、木犀草素、汉黄芩素与关键靶点ATK1、TNF、TP53具有良好亲和力。结论本研究初步明确宣肺润燥解毒方通过多成分-多靶点-多通路治疗新冠肺炎的作用机制,为深入研究宣肺润燥解毒方治疗新冠肺炎的作用机制提供了前期参考。