基于网络药理学及分子对接的五子衍宗丸治疗肝纤维化作用机制探讨

目的 基于网络药理学和分子对接技术探讨补肾复方五子衍宗丸治疗肝纤维化的作用机制。方法 使用中药系统药理学数据库与分析平台(TCMSP,http://lsp.nwu.edu.cn/tcmspsearch.php)数据库预测五子衍宗丸有效成分的靶点并通过Uniprot数据库进行靶点名称标准化;通过OMIM、Gene Cards、DisGeNET数据库收集肝纤维化靶点,通过韦恩图筛选活性成分靶点和疾病靶点基因的交集,获取五子衍宗丸治疗肝纤维化的潜在靶点;运用Cytoscape 3.9.0构建有效成分-靶点-肝纤维化网络,预测核心靶点,构建基于STRING平台的蛋白质-蛋白质相互作用(PPI)网络;利用微生信平台进行基因本体(GO)功能和京都基因与基因组百科全书(KEGG)通路富集分析;使用AutoDockTools软件,结合PPI网络及GO功能、KEGG通路分析,将核心靶点与有效成分进行对接验证。结果 获得五子衍宗丸活性成分84个,筛选出有效成分作用靶点179个,肝纤维化靶点798个,五子衍宗丸与肝纤维化交集靶点81个,经拓扑属性分析筛选得到五子衍宗丸26个活性成分、37个治疗肝纤维化关键靶点;根据“活性成分-靶点-通路”网络图,预测槲皮素、山柰酚、苦参碱、黄豆黄素为五子衍宗丸作用于肝纤维化的重要活性成分;RAC-α丝氨酸/苏氨酸蛋白激酶(AKT1)、肿瘤坏死因子(TNF)、白细胞介素-6(IL6)、细胞肿瘤抗原p53(TP53)、血管内皮生长因子A(VEGFA)等为核心靶点。GO功能富集分析的生物学过程(BP)主要与RNA聚合酶II启动子转录的正调控、基因表达的正调控等关联;细胞组分(CC)主要与细胞外空间、细胞核等关系密切;分子功能(MF)主要与大分子复合物结合、蛋白激酶活性、RNA聚合酶II核心启动子近端序列特异性DNA结合等相关。五子衍宗丸治疗肝纤维化途径主要包括化学致癌-活性氧信号通路、JAK-STAT信号通路、PI3K-Akt信号通路、MAPK信号通路等。核心成分槲皮素、山柰酚、苦参碱、黄豆黄素与核心作用靶点AKT1、TNF、IL6、VEGFA、CASP3等对接结合稳定,分子对接初步证明关键成分自发地与多个核心蛋白结合,可对多个关键靶点进行调控。结论 通过网络药理学及分子对接初步揭示补肾复方五子衍宗丸影响凋亡过程的正负调控、氧化应激、炎症因子、血管生成等反应过程,具有多成分、多靶点、多通路等作用特点,可为临床肝纤维化治疗中重视补肾法提供参考依据。     

基于网络药理学和分子对接技术探讨枳实治疗肝内胆汁淤积的作用机制

目的:运用网络药理学和分子对接技术探讨枳实治疗肝内胆汁淤积的有效活性成分及其潜在作用机制,并对枳实黄酮类粗提物治疗胆汁淤积的药效进行验证。方法:从TCMSP数据库和中国知网文献中检索出枳实的有效活性成分并收集其靶点;通过GeneCards和OMIM数据库收集肝内胆汁淤积相关的疾病靶点,将与药物靶点重复的靶点作为潜在靶点,使用Cytoscape构建“药物-有效成分-靶点-疾病”网络,利用String数据库构建蛋白互作网络(PPI);借助Metascape数据库进行GO富集与KEGG通路富集分析,最后利用AutoDock vina进行分子对接进行初步验证。观察枳实黄酮粗提物对胆汁淤积大鼠谷丙转氨酶(ALT)、谷草转氨酶(AST)、总胆汁酸(TBA)、总胆红素(TBIL)、直接胆红素(DBIL)和碱性磷酸酶(ALP)指标的影响。结果:筛选得到35个活性成分,相关潜在靶点161个,疾病靶点1 248个,枳实治疗肝内胆汁淤积潜在靶点87个,主要作用于AKT1、TP53、TNF、CASP3、VEGFA、IL6、HIF1A等19个核心靶点,这些共同作用靶点富集于4 405个生物过程和246条信号通路...     

基于网络药理学和分子对接探讨参苓白术散治疗代谢相关脂肪性肝病作用机制

目的 应用网络药理学方法和分子对接探讨参苓白术散治疗代谢相关脂肪性肝病的潜在作用机制。方法 根据TCMSP数据库搜集中药活性成分及其作用靶点,GeneCards、Drugbank数据库搜集疾病靶点,借助Cytoscape 3.9.1软件预测关键活性成分及核心靶点。交集靶点通过STRING数据库进行蛋白互作分析,Metascape数据库进行GO功能和KEGG通路富集分析,最后借助AutoDock Vina 1.1.2软件进行分子对接验证。结果 筛得关键活性成分有槲皮素、豆甾醇、山柰酚、木犀草素、β-谷甾醇、7-甲氧基-2甲基异黄酮、柚皮素等,核心靶点有AKT1、TNF、IL6、TP53、VEGFA、IL1B、CASP3、ESR1、MAPK3、PTGS2,潜在作用靶点主要富集在P53、TNF、IL17、Th17细胞分化等信号通路。分子对接结果显示结合力均良好。结论 参苓白术散治疗代谢相关脂肪性肝病具有多成分、多靶点、多通路的特点。     

蒙药安神补心六味丸抗抑郁作用网络药理学机制研究北大核心CSCD

目的应用网络药理学、分子对接技术、体外实验技术,预测以及验证蒙药安神补心六味丸的抗抑郁药理作用机制。方法TCMSP数据库及文献挖掘获取安神补心六味丸的活性成分,SwissTargetPrediction预测成分相关靶点信息,Cytoscape构建药材-成分-靶点网络。通过GeneCards、Drugbank、OMIM等数据库检索抑郁症靶点。STRING数据库获取蛋白相互作用关系网络信息。DAVID数据库进行GO生物过程富集分析和KEGG信号通路富集分析。Autodock vina软件进行分子对接验证。BV2小胶质细胞建立体外损伤模型,CCK-8法检测细胞活力,qPCR法检测相关核心靶点mRNA表达量。结果筛选出安神补心六味丸活性成分34个,涉及140个潜在作用靶点,筛选核心靶点59个,共涉及99条信号通路;分子对接结果显示桦木酸、豆甾醇、β-谷固醇、槲皮素等10种活性成分与AKT1、APP、ALB、MAPK3、VEGFA、MAPK1靶点均拥有良好的结合能力。体外实验结果表明,经给药后,BV2细胞活力较模型组明显上升。安神补心六味丸可调节各核心靶点mRNA的表达。结论安神补心六味丸可能主要通过羟色胺突触等信号通路以及相关核心靶点来发挥抗抑郁作用。     

基于网络药理学和分子对接预测红花绿绒蒿活性成分抗肝纤维化的作用机制

目的 运用网络药理学和分子对接技术,研究红花绿绒蒿抗肝纤维化的作用机制。方法 通过TCMSP、SwissTarget和PubChem数据库获取红花绿绒蒿的主要有效成分及作用靶点,从GeneCards和OMIM数据库获取肝纤维化相关疾病靶点,然后利用Venny2.1获取两者交集的共同靶点,将其导入STRING数据库构建共同靶点的蛋白互相作用网络,应用Cytoscape3.6.1软件构建“成分-靶点-疾病”通路网络。最后采用AutoDocktools(v1.5.6)将核心成分与关键靶点进行分子对接验证。结果 获取117个有效成分的作用靶点与6 423个疾病靶点,两者取交集后共获得251个交集靶点;GO与KEGG富集分析显示红花绿绒蒿有效成分可能参与AGE-RAGE、化学致癌-受体激活、PI3K-AKT等信号通路来逆转肝纤维化;分子对接显示,红花绿绒蒿核心成分与关键靶点SRC、HSP9OAA1、MAPK1和MAPK3具有较好的结合能。结论 红花绿绒蒿可能作用于SRC、HSP9OAA1、MAPK1和MAPK3等关键靶点,通过AGE-RAGE、脂质与动脉粥样硬化、化学致癌-受体激活和PI3K-A...     

基于网络药理学与分子对接技术探讨黄芪治疗鼻炎的作用机制

目的：运用网络药理学与分子对接技术探讨黄芪治疗鼻炎的分子作用机制。方法：通过TCMSP、Swiss Target Prediction、Similarity ensemble approach、Pharm Mapper数据库挖掘黄芪的活性成分及作用靶点,通过Gene Cards数据库筛选鼻炎相关靶点,将两者靶点取交集,得到黄芪治疗鼻炎的预测靶点;运用Cytoscape软件构建可视化“活性成分-靶点”网络;应用STRING数据库和Cytoscape软件构建靶点蛋白互作网络（PPI）;使用R软件对靶点进行GO功能富集分析和KEGG通路富集分析;运用Cytoscape软件构建可视化“通路-靶点”网络;通过vina软件将筛选的主要活性成分与核心靶点进行结果验证。结果：筛选得到黄芪相关活性成分16个,涉及靶点147个,其中主要活性成分包括槲皮素、山奈酚、芒柄花黄素、异鼠李素等,关键靶点包括TP53、AKT1、TNF、IL-6、VEGFA等;GO功能富集分析共得到2385条生物过程（BP）,75条细胞组分（CC）,140条分子功能（MF）;KEGG通路富集分析共得到166条信号通路,涉及动脉粥样硬...     

基于网络药理学和分子对接技术探讨温阳抗寒汤治疗支气管哮喘的作用机制

目的 采用网络药理学和分子对接技术探讨温阳抗寒汤治疗支气管哮喘的潜在作用机制。方法 基于中药系统药理学数据库与分析平台TCMSP获取温阳抗寒汤有效活性成分及其靶点,通过Gene Cards、OMIM、Pharm GKB、TTD、Drug Bank数据库获取哮喘相关靶点,将药物靶点与疾病靶点映射得到温阳抗寒汤治疗支气管哮喘的预测靶点。使用Cytoscape软件绘制药物活性成分-靶点网络图,使用String数据库构建蛋白互作网络,使用R语言进行GO富集分析和KEGG通路富集分析,预测温阳抗寒汤治疗支气管哮喘的作用机制,使用Auto Dock Tools、Autodock vina、Pymol软件进行分子对接预测和部分对接结果可视化。结果 筛选得到温阳抗寒汤作用于支气管哮喘的有效活性成分115个,检索得到支气管哮喘相关的靶点4 802个,温阳抗寒汤与支气管哮喘的交集靶点86个,关键有效成分包括槲皮素、山奈酚、木犀草素、汉黄芩素、黄芩苷、柚皮素等,涉及的核心靶点包括HIF1A、TP53、AKT1、ESR1等。通过GO分析（P <0.05）得到GO条目1 866个,其中包括生物过程（BP）...     

基于网络药理学探讨板蓝根抗呼吸道感染的相关机制

目的 基于网络药理学方法探讨板蓝根治疗呼吸道感染可能有关的相关机制研究。方法 通过TCMSP数据库筛选出板蓝根潜在活性成分,通过Gene Cards、OMIM和Drugbank数据库来预测呼吸道感染的潜在靶点;应用David和String数据库对潜在核心靶点进行基因功能富集分析和京都基因与基因组百科全书富集分析,并对富集结果进行可视化;运用String数据库和Cytoscape软件构建蛋白间相互作用网络和“通路-靶点-成分”关系网络。利用软件对核心靶点和有效成分进行分子对接。结果 板蓝根治疗呼吸道感染的核心靶点Degree值前列的有：胰岛素（INS）、血管内皮生长因子A(VEGFA)、蛋白激酶B-α(AKT1)、肿瘤蛋白p53(TP53)、白介素-6(IL-6)、过氧化物合酶2(PTGS2)、丝裂原活化蛋白激酶1(MAPK1)、丝裂原活化蛋白激酶14(MAPK14)。KEGG通路富集主要的通路有：TNF信号通路、P53信号通路、缺氧诱导因子-1信号通路、PI3K-Akt信号通路、癌症通路等。分子对接结果发现,靛蓝、靛玉红等双吲哚类生物碱跟核心靶点有较好的亲和力。结论 本研究初步揭示了板...     

基于网络药理学和反向分子对接的西红花抗肿瘤作用机制研究

目的 运用网络药理学和反向分子对接技术探讨西红花抗肿瘤的作用机制。方法 通过检索文献和从中药系统药理学数据库（TCMSP）获取西红花的主要化学成分,利用PharmMapper服务器预测西红花活性成分的潜在靶点,通过UniProt数据库将西红花潜在靶点转换为对应的靶标基因,与GeneCards、OMIM和TTD数据库获得的肿瘤相关靶点进行映射分析,得到西红花抗肿瘤的潜在作用靶点。采用Cytoscape软件构建西红花“活性成分-靶点-疾病”相互作用网络,通过String数据库进行蛋白-蛋白相互作用分析,并利用Metascape平台进行GO功能和KEGG通路富集分析。使用AutoDock、Pymol软件对活性成分与关键靶点进行分子对接验证。结果 共筛选出西红花中槲皮素、山柰酚、异鼠李素、苦番红花素和西红花苷Ⅰ等9个活性成分,可能作用于AKT1、CCND1、MMP9、EGFR、TP53等37个关键靶点,涉及P53信号通路及TNF信号通路等。分子对接显示可通过氢键、疏水作用等产生稳定结合。结论 初步探讨了西红花抗肿瘤的主要活性成分、关键靶点及通路,提示可通过诱导细胞凋亡等方式发挥抗肿瘤作用,为后...     

藏药三果汤治疗糖尿病作用机制的网络药理学与分子对接技术分析

目的 基于网络药理学及分子对接对三果汤治疗糖尿病靶点及通路进行分析，并通过分子对接进行验证，为基础研究及临床用药提供科学依据。方法 通过中药系统药理学数据库与分子平台（TCMSP）、有机小分子生物活性数据库（PubChem）及文献查阅筛选三果汤中诃子、毛诃子、余甘子的活性成分及作用靶点，使用Genecard数据库、DRUGBANK数据库筛选糖尿病相关靶点，构建“药物-成分-靶点-疾病”网络，取药物疾病靶点交集，使用STRING构建蛋白PPI网络，运用Matescape数据库对交集靶点基因进行GO、KEGG富集分析。将筛选出的有效成分与靶点进行分子对接验证。结果 通过网络药理学收集三果汤及糖尿病靶点信息，采用GO富集分析和KEGG通路分析发现，三果汤和糖尿病共同作用靶点共269个，靶点作用最为突出的为AKT1、HAS2、MAP2、CDH1、PRKCG、PLAT，三果汤作用于糖尿病的通路主要在癌症通路、脂质和动脉粥样硬化、MAPK信号通路。分子对接结果显示，鞣花酸和没食子酸对AKT1、HAS2、MAP2存在亲和力，鞣花酸对HAS2亲和力最好。结论 通过本研究中网络药理学分析及分子对接分析，三果汤中鞣花酸、没食子酸对AKT1、HAS2、MAP2通路靶点具有亲和力，为后续实验提供研究思路。     

Efficacy of gut lavage,hemodialysis, and hemoperfusion in the therapy of paraquat or diquat intoxication

Clinical and in vitro investigations were carried out to test the efficacy of gut lavage, hemodialysis, and hemoperfusion in the treatment of poisoning with paraquat or diquat. In a patient suffering from diquat intoxication 130 times more diquat was removed by gut lavage 30 h after ingestion than was removed by complete aspiration of the gastric contents.Determination of in vitro clearances for paraquat and diquat by hemodialysis showed that, at serum concentrations of 1–2 ppm, such as are frequently encountered in poisoning in man, toxicologically relevant quantities of herbicide cannot be removed from the body. At a concentration of 20 ppm, on the other hand, hemodialysis proved to be effective, the clearance being 70 ml/min at a blood flow rate of 100 ml/min. The efficacy of hemoperfusion with coated activated charcoal was on the whole better. Especially at concentrations around 1–2 ppm, the clearance values for hemoperfusion were some 5–7 times higher than those for hemodialysis.In a patient suffering from paraquat poisoning, both hemodialysis as well as hemoperfusion were carried out. The in vitro results could be confirmed: At serum concentrations of paraquat less than 1 ppm no clearance could be obtained by hemodialysis while by hemoperfusion with activated charcoal quite high clearance values were measured and the serum level dropped down to zero.

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Zusammenfassung Klinische Untersuchungen und Laboratoriumsversuche wurden durchgeführt, um die Wirksamkeit von Darmspülung, Hämodialyse und Hämoperfusion bei Paraquat- und Deiquat-Vergiftungen zu prüfen.Bei einem Patienten wurde 30 Std nach Deiquat-Aufnahme durch Darmspülung 130mal mehr Deiquat entfernt als durch vollständige Aspiration des Mageninhaltes. In vitro-Versuche ergaben, daß bei Blutserumkonzentrationen von 1–2 ppm, die bei Vergiftungen oft gemessen werden, durch Hämodialyse keine toxikologisch relevanten Paraquat- oder Deiquat-Mengen entfernt werden können. Dagegen erwies sich die Hämodialyse bei 20 ppm und einer Blutumlaufgeschwindigkeit von 100 ml/min mit einer Clearance von 70 ml/min als wirksam. Die Hämoperfusion mit beschicheter Aktivkohle war in diesen Versuchen aber eindeutig überlegen, denn insbesondere bei Konzentrationen um 1–2 ppm waren die Clearance-Werte 5–7mal höher als bei der Hämodialyse.Die in vitro-Ergebnisse wurden bei einem Patienten mit einer Paraquat-Vergiftung bestätigt: Bei Konzentrationen unter 1 ppm war die Hämodialyse wirkungslos, während durch Hämoperfusion relativ hohe Clearance-Werte erreicht wurden, so daß der Serumspiegel rasch unter die Nachweisgrenze abfiel.

     

In vitro studies on sterically stabilized liposomes (SL) as enzyme carriers in organophosphorus (OP) antagonism

This study describes a new approach for organophosphorous (OP) antidotal treatment by encapsulating an OP hydrolyzing enzyme, OPA anhydrolase (OPAA), within sterically stabilized liposomes. The recombinant OPAA enzyme was derived from Alteromonas strain JD6. It has broad substrate specificity to a wide range of OP compounds: DFP and the nerve agents, soman and sarin. Liposomes encapsulating OPAA (SL)* were made by mechanical dispersion method. Hydrolysis of DFP by (SL)* was measured by following an increase of fluoride ion concentration using a fluoride ion selective electrode. OPAA entrapped in the carrier liposomes rapidly hydrolyze DFP, with the rate of DFP hydrolysis directly proportional to the amount of (SL)* added to the solution. Liposomal carriers containing no enzyme did not hydrolyze DFP. The reaction was linear and the rate of hydrolysis was first order in the substrate. This enzyme carrier system serves as a biodegradable protective environment for the recombinant OP-metabolizing enzyme, OPAA, resulting in prolongation of enzymatic concentration in the body. These studies suggest that the protection of OP intoxication can be strikingly enhanced by adding OPAA encapsulated within (SL)* to pralidoxime and atropine.     

Aquatic Insects and Trace Metals: Bioavailability,Bioaccumulation, and Toxicity

Abstract

The uptake of metals from food and water sources by insects is thought to be additive. For a given metal, the proportions taken up from water and food will depend both on the bioavailable concentration of the metal associated with each source and the mechanism and rate by which the metal enters the insect. Attempts to correlate insect trace metal concentrations with the trophic level of insects should be made with a knowledge of the feeding relationships of the individual taxa concerned. Pathways for the uptake of essential metals, such as copper and zinc, exist at the cellular level, and other nonessential metals, such as cadmium, also appear to enter via these routes. Within cells, trace metals can be bound to proteins or stored in granules. The internal distribution of metals among body tissues is very heterogeneous, and distribution patterns tend to be both metal and taxon specific. Trace metals associated with insects can be both bound on the surface of their chitinous exoskeleton and incorporated into body tissues. The quantities of trace meals accumulated by an individual reflect the net balance between the rate of metal influx from both dissolved and particulate sources and the rate of metal efflux from the organism. The toxicity of metals has been demonstrated at all levels of biological organization: cell, tissue, individual, population, and community. Much of the literature pertaining to the toxic effects of metals on aquatic insects is based on laboratory observations and, as such, it is difficult to extrapolate the data to insects in nature. The few experimental studies in nature suggest that trace metal contaminants can affect both the distribution and the abundance of aquatic insects. Insects have a largely unexploited potential as biomonitors of metal contamination in nature. A better understanding of the physico-chemical and biological mechanisms mediating trace metal bioavailability and exchange will facilitate the development of general predictive models relating trace metal concentrations in insects to those in their environment. Such models will facilitate the use of insects as contaminant biomonitors.     

Alzheimer’s disease – current trends in basic and clinical research. Highlights from the 16th ECNP

Advances in the molecular biological knowledge of neuronal nicotinic acetylcholine receptors (nAChRs) have led to a growing interest by the pharmaceutical industry in the development of novel compounds that selectively modulate nAChR function. The ability of (-)-nicotine, an activator of nAChRs, to enhance attentional aspects of cognition in animals and humans, to exert neuroprotective and anxiolytic-like effects, and presumably to mediate the negative correlation between smoking and Alzheimer's (and Parkinson's) Disease, has focused interest on the potential therapeutic utility of modulators of nAChR function for treatment of some of the deficits associated with these progressive, neurodegenerative conditions. Numerous compounds are known which activate nAChRs and which might serve as lead compounds toward the development of such agents. The pharmacologic diversity of neuronal nAChR subtypes suggests the possibility of developing selective compounds which would have more favourable side-effect profiles than existing agents. This broader class of agents, collectively called cholinergic channel modulators (ChCMs), is anticipated to encompass compounds which would have more favourable side-effect profiles than existing agents, which generally exhibit low selectivity. This selectivity may be achieved by preferentially activating some subtypes of nAChRs (i.e., Cholinergic Channel Activators, ChCAs) or inhibiting the function of other subtypes (Cholinergic Channel Inhibitors, ChCIs). An overview of the biology of nAChRs and the rationale for the use of ChCMs for the treatment of dementia related to neurodegenerative diseases are presented, followed by a discussion of lead compounds and compounds under consideration for clinical evaluation.     

Steroidal sapogenin contents in some domestic plants

In order to find out the values of the steroid resources for the future use. the compositions and contents of steroidal sapogenins from 13 domestic plants have been investigated. As a result,Dioscorea nipponica, D. quinqueloba andSmilax china were found to have large amount of diosgenin. And pennogenin inTrillium kamtschaticum andParis verticillata, yuccagenin inAllium fistulosum, hecogenin inAgave americana and neochlorogenin inSolanum nigum were appeared to be major steroidal sapogenins.