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.

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.

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。

基于网络药理学和分子对接技术的调压方治疗原发性高血压的作用机制

目的通过网络药理学方法和分子对接技术探究调压方治疗原发性高血压的作用机制。方法利用TCMSP、TCM中药数据库及文献获取中药成分;Super-PRED分析药物活性成分的靶点;通过GeneCards、NCBI、OMIM、PharmGKB疾病数据库检索获取原发性高血压的疾病靶点;选择药物—疾病交集靶点通过Cytoscape软件分析获取药物核心作用靶点和核心成分;利用DAVID数据库完成核心靶点的GO和KEGG富集分析;通过CB-Dock2在线分子对接调压方核心成分与核心靶点。结果筛选获取得到调压方药物活性成分284个;调压方活性成分靶点311个;原发性高血压疾病靶点1550个;药物—疾病交集靶点107个;核心作用靶点NFKB1、KDR、STAT3、TP53、PTGS2、TLR4等19个;GO富集分析显示,调压方通过多种生物学过程参与抗原发性高血压;KEGG富集分析显示,调压方通过癌症蛋白聚糖、脂质和动脉粥样硬化等途径调控原发性高血压;分子对接结果显示,调压方核心成分与核心靶点之间有较好的结合活性。结论调压方可通过“多成分—多靶点-多途径”方式作用于NFKB1、KDR、STAT3、CXCR4、TLR4、SRC等靶点参与脂质与动脉粥样硬化信号通路、MAPK信号通路、PI3K-AKT信号通路等途径发挥治疗原发性高血压作用。

中药益气血方对超促排卵小鼠卵巢生长分化因子表达的影响

【目的】研究中药益气血方对超促排卵小鼠卵巢生长分化因子-9(growth differentiation factor-9,GDF-9)与GDF-9B表达的影响。【方法】选用昆明种小鼠接受促性腺激素释放激素类似物(GnRHa)与孕马血清(PMSG)超促排卵,中药组予以益气血方(经后增殖颗粒)(剂量为1.5 mg/kg)灌胃,对照组予以生理盐水灌胃。注射绒毛膜促性腺激素(hCG)后取卵巢组织,采用免疫组织化学方法检测GDF-9与GDF-9B的表达与半定量分析,采用实时荧光定量PCR方法检测GDF-9、GDF-9B mRNA的含量。【结果】卵母细胞与颗粒细胞中检测到GDF-9的表达,GDF-9B未表达于卵母细胞,而在颗粒细胞中高表达。半定量分析结果显示中药组卵巢组织中GDF-9与GDF-9B蛋白表达均显著高于对照组(P<0.05),实时荧光定量PCR结果显示中药组GDF-9、GDF-9B mRNA表达水平均显著高于对照组(P<0.05)。【结论】中药益气血方可促进超促排卵小鼠卵巢GDF-9与GDF-9B的表达,可能通过此机制促进卵母细胞发育潜能,从而促进胚胎种植。

益气血法对超排卵小鼠卵巢颗粒细胞Bcl-2、Bax、Caspase3表达的影响

目的观察经后增殖方对超排卵小鼠卵巢凋亡相关基因及其蛋白表达的调节作用,探讨中药益气血法对超排卵小鼠卵巢颗粒细胞凋亡及卵泡质量的影响。方法建立超排卵小鼠模型,30只小鼠随机分为空白组、模型组、中药治疗组。采用实时荧光定量PCR(real-time quantitative PCR,qPCR)和免疫印迹法(Western blot)分别检测Bcl-2、Bax、Caspase3基因和蛋白的表达水平。结果中药治疗组Bax、Cas-pase3 mRNA的表达水平与模型组比较(0.895 vs 1.779,0.674 vs 2.066,均P<0.05),差异均有统计学意义;中药治疗组Bcl-2、Bax、Caspase3 mRNA的表达水平与空白组比较(1.11 vs 1.26,0.89 vs 0.79,0.67 vs 0.96),差异均无统计学意义(P>0.05);中药治疗组Bcl-2、Bax、Caspase3蛋白的表达水平与模型组比较(0.62 vs0.41,0.79 vs 1.06,0.49 vs 0.89),差异均有统计学意义(P<0.05),与空白组比较(0.62 vs 0.64,0.79 vs 0.73,0.49 vs 0.50)差异均无统计学意义(P>0.05)。结论经后增殖方能促进Bcl-2蛋白的表达,抑制Bax、Cas-pase3基因及蛋白的表达,抑制超排卵小鼠卵巢颗粒细胞凋亡至接近自然排卵小鼠的水平,提高超排卵小鼠卵泡质量。

益气血补肝肾方对胚胎着床障碍小鼠子宫内膜整合素β3和胞饮突表达的影响

【目的】观察中药益气血补肝肾方对胚胎着床障碍小鼠子宫内膜整合素β3表达的影响,探讨其改善胚胎着床的分子基础。【方法】将90只雌鼠随机分为正常组、模型组和治疗组3组,每组30只,其中,治疗组小鼠予以中药益气血补肝肾方治疗(包括经后增殖方8 mg/kg和促黄体方8 mg/kg),在妊娠第4天(Pd4)上午8∶00采用米非司酮诱导模型组和治疗组小鼠胚胎着床障碍,12 h后处死小鼠(每组20只),剖腹取小鼠子宫,小心刮取子宫内膜,采用Real-time PCR和Western Blot的方法分别在m RNA和蛋白水平上检测整合素β3在各组小鼠子宫内膜中的表达情况。其余小鼠(每组10只)于Pd4剖腹取小鼠子宫,固定于体积分数2.5%戊二醛溶液中,行扫描电镜观察。【结果】Real-time RT-PCR和Western Blot结果显示:模型组整合素β3 m RNA与蛋白表达显著低于正常组(P<0.05);治疗组可显著升高整合素β3 m RNA与蛋白表达水平,与模型组比较,差异均有统计学意义(P<0.05)。胞饮突在模型组中的表达少于治疗组和正常组,而胞饮突在治疗组和正常组的表达相似。【结论】中药益气血补肝肾方可上调子宫内膜中整合素β3的表达,对胞饮突在Pd4小鼠子宫内膜中的表达有正调节作用。

益气血补肝肾中药对卵巢过度刺激综合征患者血管内皮生长因子的影响

目的观察益气血补肝肾中药对卵巢过度刺激综合征(OHSS)高危患者血管内皮生长因子(VEGF)的影响。方法选取2016年6-11月在广州军区广州总医院生殖医学中心行试管婴儿助孕的有OHSS高危因素的患者为研究对象,随机分为中药组148例和对照组150例,另选取同期145例无OHSS高危因素的患者作为正常组。所有患者均予长方案治疗,中药组在此基础上加用中药。结果中药组比对照组有更高的优质胚胎率及更低的中重度OHSS发生率,差异有统计学意义(P<0.05)。对照组和中药组在各时间点血清VEGF水平均比正常组高(P<0.05),而中药组移植日血清VEGF水平显著低于对照组(P<0.05)。结论血清VEGF水平升高与OHSS的发生密切相关,益气血补肝肾中药可降低OHSS高危患者移植后的血清VEGF水平,对中重度OHSS的发生起到预防作用。