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.

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.

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.

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.

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。

威灵仙痛风方治疗禽痛风的网络药理学分析及分子对接预测

为预测威灵仙痛风方治疗禽痛风的效果及探究其作用机制,试验采用网络药理学和分子对接的方法对威灵仙痛风方治疗禽痛风的靶点进行预测,并评估其准确性。结果表明:威灵仙复方有效成分158个,相应的作用靶点有66个;禽痛风的作用靶点1231个,二者互作潜在靶点共33个,其中核心靶点有STAT3、VEGFA、IL6、BCL2L1、IL10、PTGS2、MYC、IFNG和NFKBIA,富集到MAPK、Toll样受体、FoxO和VEGF信号通路等。威灵仙痛风方中的主要活性成分包括槲皮素、山奈酚和β-谷甾醇等,可能靶向作用于PTGS2、IL6、BCL2L1等靶点,经MAPK、Toll样受体等炎性信号通路发挥治疗效用。

基于网络药理学探讨宣肺清热方治疗COVID-19的分子机制

目的 基于网络药理学探讨宣肺清热方治疗新型冠状病毒感染(COVID-19)的潜在作用机制。方法 通过中药系统药理学数据库和分析平台(TCMSP)数据库获取宣肺清热方的活性成分及相关作用靶点;通过基因卡片(GeneCards)、在线《人类孟德尔遗传》(OMIM)、药物银行(DrugBank)数据库获取COVID-19的主要作用靶点;获得宣肺清热方与COVID-19共同靶点并制作韦恩图(Venn Diagram);利用基因/蛋白质互相作用检索分析工具(STRING)进行蛋白质-蛋白质相互作用(protein-protein interaction,PPI)分析,构建PPI网络;通过Metascape平台进行基因本体论(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,采用Cytoscape 3.9.0软件构建“宣肺清热方-活性成分-共同靶点-COVID-19-信号通路”网络。结果 经筛选后获得宣肺清热方活性成分121个、靶点232个,COVID-19靶点2 255个及药物-疾病共同靶点68个,度值(Degree)最大的10个活性成分为槲皮素、山奈酚、木犀草素、β-谷甾醇、豆甾醇、柚皮素、异鼠李素、汉黄芩素、黄芩素和光甘草定,GO及KEGG富集分析结果主要涉及炎症-免疫调节、胞内信号转导、基因调控等多种生物学途径以及癌症通路、白细胞介素-17(IL-17)信号通路、腺苷酸活化蛋白激酶(AMPK)信号通路、钙信号通路、p53信号通路、过氧化物酶增殖物激活受体(PPAR)信号通路等。共同靶点根据度值排名,前10位是雄激素受体(AR)、一氧化氮合酶2(NOS2)、电压门控钠通道5α(SCN5A)、RELA、蛋白激酶B(AKT)1、B淋巴细胞瘤2(BCL2)、肿瘤坏死因子(TNF)、丝裂原激活蛋白激酶1(MAPK1)、核因子κB抑制剂α(NFKBIA)、白细胞介素-6(IL-6)。结论 宣肺清热方中主要化学成分可能作用于多靶点多通路介导炎症-免疫调节等生物学过程,发挥治疗COVID-19的作用。

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。

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.

祛风通络方对大鼠系膜细胞增殖及转化生长因子β1和白细胞介素6 mRNA表达的影响

目的:观察祛风通络方对脂多糖(lipopolysaccharide,LPS)诱导的大鼠肾小球系膜细胞(mesangial cell,MC)增殖及其分泌的转化生长因子β1(transforming growth factor-beta1,TGF-β1)和白细胞介素6(interleukin-6,IL-6)mRNA表达的影响。方法:采用血清药理学方法,体外培养大鼠肾小球系膜细胞,分为正常对照组、模型组、蒙诺(福辛普利钠)组和祛风通络方组。并采用甲基噻唑基四唑(methyl thiazolyl tetrazolium,MTT)法检测大鼠MC增殖情况,逆转录聚合酶链反应法检测TGF-β1和IL-6mRNA的表达。结果:与正常对照组比较,模型组大鼠肾小球系膜细胞增殖升高(P<0.05);与模型组比较,祛风通络方组大鼠肾小球系膜细胞增殖受到抑制(P<0.05),且祛风通络方作用优于蒙诺(P<0.05)。模型组系膜细胞TGF-β1 mRNA表达高于正常对照组(P<0.01);祛风通络方组和蒙诺组系膜细胞TGF-β1 mRNA表达低于模型组(P<0.01);祛风通络方组与蒙诺组比较,TGF-β1 mRNA的表达降低(P<0.01)。LPS刺激后可提高大鼠系膜细胞内IL-6 mRNA的表达,与正常对照组比较,模型组IL-6 mRNA表达上升,差异有统计学意义(P<0.01);祛风通络方组、蒙诺组与模型组比较,IL-6 mRNA表达下降(P<0.01);祛风通络方降低IL-6 mRNA表达的作用亦优于阳性对照药蒙诺(P<0.01)。结论:祛风通络方对大鼠MC增殖及其TGF-β1和IL-6 mRNA表达均有明显的抑制作用。

补肾调冲方含药血清对高雄激素培养的卵巢颗粒细胞IGF-1 StAR mRNA表达的影响

目的:运用血清药理学方法观察补肾调冲方对高雄激素培养的大鼠卵巢颗粒细胞(GC)IGF-1、StARmRNA表达的影响。方法:根据血清药理学方法,用6周龄的未成年SD大鼠制备FSH及大、中、小剂量补肾调冲方含药血清,分别加入到过量丙酸睾丸酮处理的GC培养体系中。培养48h后,用实时荧光定量RT-PCR(Real-timeRT-PCR)方法检测GCIGF-1、StARmRNA的表达量。结果:高雄激素抑制GCIGF-1、StARmRNA的表达,补肾调冲方及FSH含药血清可拮抗高雄激素的抑制作用,促进IGF-1、StARmRNA的表达。结论:补肾调冲方含药血清可有效拮抗高雄激素对IGF-1、StARmRNA表达的抑制作用,推测补肾调冲方通过促进IGF-1、StARmRNA的表达,改善雌/雄激素比值来调节卵巢功能。

基于数据挖掘和网络药理学的新冠肺炎预防方药组方规律和作用机制研究

目的基于数据挖掘和网络药理学方法,研究2019新型冠状病毒肺炎官方公布预防方药组方规律和作用机制,为新冠肺炎的防治提供指导。方法采用数据挖掘和关联规则分析方法,研究新冠肺炎预防方药组方规律;进一步采用网络药理学方法,通过KEGG通路和基因本体富集分析,研究高频核心药物的生物信号通路,阐明其作用机制。结果筛选官方公布的新冠肺炎预防方剂和中成药63个,发现14味高频中药,进一步关联分析得到14个高频药对和15个高频药组。在此基础上,采用网络药理学方法,研究高频核心药组作用机制,发现"防风、白术和黄芪"主要通过干预氨基酸代谢、嘌呤代谢等基础代谢途径发挥预防作用,而"藿香和苍术"除基础代谢外,还通过参与炎症相关反应发挥预防治疗作用。结论新冠肺炎预防方药的核心药组"防风、白术和黄芪"和"藿香和苍术"分别通过基础代谢和炎症相关通路发挥预防疗效,为主动健康指导方针下的"未病先防"提供参考。

方剂组织药理学新假说

方剂的作用特点是多成分、多途径、多环节、多靶点,其所含药效物质是方剂研究的核心,探索中药方剂的药效物质基础是中药现代化的关键之一。中药血清药理学、方剂药动学和药效学研究取得了一定的成绩,但是以进行体外研究为主,也没有深入到病变靶组织层次。以急性胰腺炎动物模型为研究对象,采用高效液相色谱-质谱联用技术与方法检测模型动物口服或灌肠后胰腺组织内的有效中药成分谱,进而提出方剂组织药理学的理论假说,即以中医基本理论为指导,用药理学的方法研究中药(方剂)有效成分对病变靶组织的影响及其作用原理的科学;建立其实验方法学,寻找中药方剂靶组织作用的基础、原理和直接药效物质基础;可能产生新的相互有效方剂,并开辟中药方剂研究新领域。

疏肝活血方含药血清对大鼠骨髓源性内皮祖细胞血管新生相关基因表达的影响

目的:观察疏肝活血方含药血清对大鼠骨髓来源的内皮祖细胞(EPCs)血管新生相关基因VEGF、HGF和HIF-1α表达的影响。方法:从大鼠骨髓中获取单个核细胞,体外培养1周后,采用免疫荧光细胞化学和激光共聚焦显微镜鉴定,于培养体系中加入疏肝活血方及其拆方的含药血清,培养24h,Real-TimePCR方法检测EPCs中VEGF、HGF和HIF-1α的mRNA表达。结果:贴壁细胞培养诱导后具有内皮细胞的形态学和免疫学特征,Real-TimePCR检测结果发现,疏肝活血方含药血清能明显提高EPCs中VEGF、HGF、HIF-1α的mRNA表达量,与对照相比差异具有显著意义(P<0.01)。结论:疏肝活血方能促进EPCs中血管新生相关基因VEGF、HGF和HIF-1α的表达,是该方促血管新生的机制之一。

补肾调冲方含药血清对高雄激素培养大鼠卵巢颗粒细胞增殖与分泌的影响

目的运用血清药理学方法观察补肾调冲方对高雄激素培养的大鼠卵巢颗粒细胞增殖与分泌功能的影响。方法根据血清药理学方法,用6周龄的未成年SD大鼠制备促卵泡生成素(FSH)及大、中、小剂量补肾调冲方含药血清,分别加入过量丙酸睾丸酮处理的颗粒细胞培养体系中。培养48h后,测定3HTdR掺入DNA的量,并用流式细胞仪检测细胞周期分布及增殖指数(PI),放免法测定颗粒细胞培养液中雌二醇(E2)、孕酮(P)水平及细胞内环磷酸腺苷(cAMP)含量。结果高雄激素明显抑制颗粒细胞的增殖与分泌功能,并明显降低了细胞内cAMP的含量;而补肾调冲方含药血清可改善高雄激素对颗粒细胞功能的抑制状态,促进细胞增殖及E2、P的分泌,其作用呈剂量依赖性。结论补肾调冲方含药血清可有效拮抗高雄激素对颗粒细胞功能的抑制作用,推测补肾调冲方通过促卵泡发育、提高雌、孕激素水平,改善雌/雄激素比值来调节卵巢功能。

参芪固本方治疗脾虚荷Lewis肺癌小鼠的实验研究

【目的】探讨参芪固本方对脾虚荷Lewis肺癌模型小鼠的治疗作用及机制。【方法】将72只小鼠随机分为4组,每组18只,分别为荷瘤模型组(A组)、脾虚荷瘤模型组(B组)、脾虚荷瘤化疗组(C组)、脾虚荷瘤中药组(D组),其中B、C、D组均以大黄煎液(30 g/kg)灌胃7 d复制脾虚模型,然后各组小鼠右侧腋窝皮下接种传代小鼠Lewis肺癌细胞混悬液(5×106/mL)0.2 mL。接种后第2天A、B组每日以生理盐水灌胃,C组以顺铂(DDP)每日腹腔注射1次(剂量为1 mg/kg),同时每日以生理盐水灌胃,D组每日以参芪固本方药液灌胃,剂量为43.83 g/kg,均连续10 d。检测模型小鼠瘤体质量,去瘤体质量,免疫器官脾、胸腺质量,并采用放射免疫法检测血清肿瘤坏死因子-α(TNF-α)、白细胞介素-6(IL-6)的水平。【结果】B组瘤体质量最重,与其他组比较差异均有显著性意义(P<0.05或P<0.01)。去瘤体质量比较,D组略高于其他组,但差异无显著性意义(P>0.05)。B、C组较A组胸腺质量减轻(P<0.01)。C、D组均可升高血清TNF-α水平(与A、B组比较,P<0.05或P<0.01),D组可显著性降低IL-6水平(与A、B组比较,P<0.01)。【结论】参芪固本方的抑瘤作用可能与其能升高血清TNF-α水平,降低IL-6水平,提高免疫功能有关。