Mechanism prediction of Astragalus mongholicus Bunge and Angelica sinensis Diels in treating interstitial lung disease based on network pharmacology and molecular docking

Objective:To investigate the mechanism by which Astragalus mongholicus Bunge(AM),and Angelica sinensis Diels(AS)act in interstitial lung disease(ILD)based on computational prediction.Methods:We screened the ingredients of AM and AS in PubMed,the Web of Science,China National Knowledge Infrastructure(CNKI)Databases,etc.Then obtained the potential effective components.By sharing the same molecular with ILD,we got the possible target genes for ILD treatment and constructed components–targets–disease network with Cytoscape software.The CTD(Comparative Toxicogenomics Database)database was used for GO and KEGG enrichment analysis of these target genes.Results:59 active ingredients that can be druggable were chosen from AM,67 active ingredients were chosen from AS.77 overlapping target genes for AM and ILD and 36 overlapping target genes for AS and ILD were acquired.The hub targets of AM were PTGS2,PTGS1,CDK2,MAOA,ESR1,TOP2A,GSK3B,ESR2,PPARG,NOS2,The hub targets of AS were PTGS2,GABRA1,PTGS1,CHRM1,SLC6A2,ADRA1B,ADRAIA,ADRB2,CHRM3,GABRA2,CHRM2.Quercetin,kaempferol,daidzein,pavilion,7-Hydroxycoumarin,and 5-Hydroxycoumarin were the main active ingredients which have more effective targets.Prediction of the protein-protein interaction network showed PTGS2,GSK3B,PPARG,etc.,were the important predicted targets.The enriched KEGG pathways,including the Immune System,Metabolism of lipids and lipoproteins,Cytokine Signaling in the Immune system,Generic Transcription Pathway,The interleukin pathway,Metabolism of proteins,PI3K-Akt signaling pathway,Metabolic pathways,Innate Immune System,Neuroactive ligand-receptor interaction,Metabolism,GPCR downstream signaling,Amine ligand-binding receptors,Class A/1,Calcium signaling pathway.Molecular docking showed that quercetin,kaempferol,daidzein,pavilion,7-Hydroxycoumarin,5-Hydroxycoumarin had good binding activities with PTGS2 and GSK3B,which mainly mediated PI3K/Akt and other important signaling pathways in the pathogenesis of ILD.Conclusion:The components in AS and AM share some common targets,such as PTGS2.AM and AS may ameliorate ILD through the PI3K-Akt signaling pathway which is mediated by GSK3B.PTGS2,PPARG may also be vital target genes in the treatment of ILD with AM and AS.

Karyotype Analysis of Angelica sinensis (Oliy.) Diels

The seeds of tested Angelica sinensis （Oliy.）Diels were purchased from the planting base of Hebsi Anguo Traditional Chinese Medicine Market. With the root tips of germinated Angelica sinensis seeds treated with GA3 as materials, karyotype analysis was performed by photomicrographic technology and conventional chromosome preparation method. The results showed that the chromosome number of A. sinensis was 2 n=2.2, which accords with previous studies; and no satellites were observed, and the chromosome karyotype was 2A, with the asymmetrical karyotype coefficient （AS.K%） of 54：85%. The total length of the chromosomes was 113.4 pro, and total volume of chromosomes was 83.21 μm3.

Mechanism prediction of Astragalus mongholicus Bunge and Angelica sinensis Diels in treating interstitial lung disease based on network pharmacology and molecular docking

Objective:To investigate the mechanism by which Astragalus mongholicus Bunge(AM),and Angelica sinensis Diels(AS)act in interstitial lung disease(ILD)based on computational prediction.Methods:We screened the ingredients of AM and AS in PubMed,the Web of Science,China National Knowledge Infrastructure(CNKI)Databases,etc.Then obtained the potential effective components.By sharing the same molecular with ILD,we got the possible target genes for ILD treatment and constructed components–targets–disease network with Cytoscape software.The CTD(Comparative Toxicogenomics Database)database was used for GO and KEGG enrichment analysis of these target genes.Results:59 active ingredients that can be druggable were chosen from AM,67 active ingredients were chosen from AS.77 overlapping target genes for AM and ILD and 36 overlapping target genes for AS and ILD were acquired.The hub targets of AM were PTGS2,PTGS1,CDK2,MAOA,ESR1,TOP2A,GSK3B,ESR2,PPARG,NOS2,The hub targets of AS were PTGS2,GABRA1,PTGS1,CHRM1,SLC6A2,ADRA1B,ADRAIA,ADRB2,CHRM3,GABRA2,CHRM2.Quercetin,kaempferol,daidzein,pavilion,7-Hydroxycoumarin,and 5-Hydroxycoumarin were the main active ingredients which have more effective targets.Prediction of the protein-protein interaction network showed PTGS2,GSK3B,PPARG,etc.,were the important predicted targets.The enriched KEGG pathways,including the Immune System,Metabolism of lipids and lipoproteins,Cytokine Signaling in the Immune system,Generic Transcription Pathway,The interleukin pathway,Metabolism of proteins,PI3K-Akt signaling pathway,Metabolic pathways,Innate Immune System,Neuroactive ligand-receptor interaction,Metabolism,GPCR downstream signaling,Amine ligand-binding receptors,Class A/1,Calcium signaling pathway.Molecular docking showed that quercetin,kaempferol,daidzein,pavilion,7-Hydroxycoumarin,5-Hydroxycoumarin had good binding activities with PTGS2 and GSK3B,which mainly mediated PI3K/Akt and other important signaling pathways in the pathogenesis of ILD.Conclusion:The components in AS and AM share some common targets,such as PTGS2.AM and AS may ameliorate ILD through the PI3K-Akt signaling pathway which is mediated by GSK3B.PTGS2,PPARG may also be vital target genes in the treatment of ILD with AM and AS.

Differentially Expressed Genes in Heads and Tails of Angelica Sinensis diels： Focusing on Ferulic Acid Metabolism

Objective： To explore the scientific connotation of the discrepant pharmaceutical activities between the head and tail of Angelica sinensis diels（AS）, an important herb extensively utilized in Chinese medicine, by the approach of transcriptome sequencing. Methods： Ten samples of AS were randomly collected in Min County, Gansu Province of China. Transcriptome sequencing of AS was accomplished in a commercial ILLumina Hi Seq-2000 platform. The transcriptome of each head and tail of AS were fixed in a gene chip, and detected under the procedure of Illumina Hi Seq-2000. Differentially expressed unigenes between the heads and tails of AS were selected by Shanghai Biotechnology Corporation（SBC） online analysis system, based on Gene Ontology, Kyoto Encyclopedia of Genes and Genomes and relevant bioinformatic database. Results： Totally 63,585 unigenes were obtained from AS by high-throughput sequencing platform. Among which 3359 unigenes were identified as differentially expressed unigenes between the heads and tails of AS by SBC analysis system scanning. Of which 15 differentially expressed unigenes participate in the metabolic regulation of phenylpropanoid biosynthesis（PB） pathway and ferulic acid metabolites, in response to the distinguished pharmaceutical actions of the heads and tails of AS. Conclusion： Different content of ferulic acid in the heads and tails of AS is related to the differentially expressed genes, particularly involved in the PB pathway.

正交试验优化当归趁鲜切片的加工工艺

目的通过药物成分含量检测与正交试验相结合的方式,确定当归趁鲜切片的工艺流程及工艺参数。方法先考察鲜当归杂质处理方法、切片的厚度及其干燥的方式对当归趁鲜切制外观性状、内在结构和药物成分(阿魏酸、挥发油)含量的影响。然后采用L_(9)(3^(4))正交试验,对当归饮片中阿魏酸含量进行分析,选出当归趁鲜切制的最适宜工艺条件。结果通过正交试验优化得到最适宜当归趁鲜饮片加工工艺为:首先将鲜当归除去芦头、须根和泥沙,并用水洗净其表面泥土和杂质,稍晾至没有水滴下滴时,放入温棚日晒干燥,直到当归归须和归尾(直径在5 mm以内)已干燥,归身变软,再将全当归切成4 mm薄片,最后放置在温棚日晒干燥,即得当归饮片,其饮片中阿魏酸含量为0.2211%,挥发油含量为0.9%;当归趁鲜切制最适宜工艺所得的当归饮片质量优于传统方法所得的质量(阿魏酸含量为0.1930%,挥发油含量为0.8%)。结论所得的加工工艺合理、可行,为后期当归趁鲜切片工艺研究奠定基础。

栽培方式对航天诱变当归SP_(2)繁种特性的影响

当归3年完成正常生活史,繁种周期长,成种株率低。为提高当归新品种选育效率,将搭载“长征7号”运载火箭在太空运行22 h(命名为航归22 h)和搭载“神舟十一号”宇宙飞船在太空运行33 d(命名为航归33 d)的当归种子作为SP0种子,以未搭载种子为对照(CK),经育苗栽培获得SP_(2)种苗。采用留苗和移栽2种栽培方式,测定进入第3年繁种年的SP_(2)群体的种株率、农艺性状和种果重。结果表明,CK群体未能进入繁种期而被淘汰,太空诱变获得的2个群体均进入正常繁种年。航归22 h的返青株成活率较航归33 d提高14.65个百分点,成种株率提高19.7个百分点。与移栽航归33 d、留苗航归22 h和33 d群体相比较,移栽航归22 h的繁殖特性有明显改善,单株果重分别增加1527.5%、428.2%和377.7%,根重分别增加235.3%、256.8%和265.2%,根长分别增加44.5%、36.7%和6.0%,根粗分别增加83.3%、83.7%和83.9%。航归SP_(2)代群体性状综评指数大小依次为移栽航归22 h(0.8133)>留苗航归33 d(0.4083)>留苗航归22 h(0.2378)>移栽航归33 d(0.0334)。综上,太空搭载种子对当归具有诱变效应,运载火箭搭载时间短,群体诱变效应更佳,选育效率更高;采用移栽和留苗互补栽培可保障繁种的安全性。本研究结果为当归新品系扩繁提供了科学技术依据。

应用Proofman-LMTIA技术双重检测当归和东当归

通过梯型熔解温度核酸等温扩增技术(Ladder-shape melting temperature nucleic acid isothermal amplification,LMTIA)与校对酶介导探针(Proofreading enzyme-mediated probe cleavage,Proofman)联合的方法对当归和东当归进行双重检测。以ddH2O为阴性模板,当归和东当归基因组DNA为阳性模板,测定当归和东当归引物的最适温度、灵敏度及稳定性,并进行了真实样品检测。结果表明,应用Proofman-LMTIA双重检测当归和东当归,引物测定的最适温度为62℃,当归引物灵敏度低至10 pg/μL,东当归引物灵敏度低至1 pg/μL,具有良好的稳定性。7份市售当归样品经检测均为当归。该研究可为当归和东当归的鉴别提供一种新的检测方法。

氨基酸螯合铁锰耦合对当归生长与品质的影响及其生理响应分析

目的考察氨基酸螯合铁锰耦合对当归生长与品质的影响,并进行生理响应分析。方法在田间试验中,设置叶面分别喷施0.02%氨基酸螯合Fe(T1)、氨基酸螯合Mn(T2)、氨基酸螯合Fe+Mn(T3)和无离子水(CK),分析了不同处理对药材生长发育状况、产量、一等归率及光合参数、抗氧化酶活性、渗透调节物质含量和药效物质含量的影响。结果与CK比较,其他3个处理均能显著促进药材植株生长,提高其产量、一等归率及阿魏酸、洋川芎内酯I、阿魏酸松柏酯、洋川芎内酯A、Z-藁本内酯含量。药材根中Fe、Mn含量呈显著负相关。Fe、Mn含量和光合参数可解释产量和一等归率变化的比例分别为21.06%、1.69%和37.07%。以5种有机药效成分积累为指标,各处理对药材质量的影响程度依次为T3>T2>T1>CK。结论氨基酸螯合铁锰耦合对当归产量、一等归率、光合作用和抗胁迫能力均有促进作用,对药材品质有增进作用,建议其施用浓度为0.2%,全生长期喷布3次。

当归药材道地性的RAPD分析

目的 从 DNA分子水平上探讨当归药材道地性的生物学基础。方法 对来自不同产地的 18个当归样品 ,提取其基因组 DNA,并进行 RAPD分析。对筛选出的 10个引物的 RAPD结果进行聚类分析 ,得出 18个当归样品的聚合树状图。结果 显示地理分布距离越小 ,当归的遗传差异越小 ;反之 ,越大。但生态环境特别是小生境对当归遗传差异的作用亦不可忽视。

当归的研究进展

对近年在当归植物学、药理学、栽培学等方面的研究工作作一概述。就当归生地育苗。

当归补血汤中不同组分对正常及血虚小鼠免疫功能的影响

目的 比较当归补血汤中的不同组分对正常及血虚小鼠模型的免疫调节作用。方法 采用 MTT法测 T、B淋巴细胞的增殖率 ;分光光度法检测脾细胞中抗体的生成 ;用乙酰苯肼和环磷酰胺制备小鼠血虚模型。结果 黄芪中的 4个不同组分对正常小鼠均具有增强 T、B淋巴细胞增殖率及促进抗体生成作用 ,而当归中的 3个不同组分则作用各异 ,但是对血虚模型小鼠均表现出明显的免疫调节作用。体外实验发现 :在 2 .5～ 2 5 0 μg/m L 浓度范围内 ,黄芪黄酮、黄芪皂苷以及黄芪和当归的挥发油部分均表现为良好的双向免疫调节作用 ,而黄芪和当归的多糖组分则为明显的浓度依赖性作用方式。

不同产地当归药材及其土壤无机元素的关联分析与探讨

目的:通过对不同产地当归药材及其土壤无机元素类成分的关联分析,以评价该类成分与药材地道性和功效间的关系。方法:采用微波消解法-电感耦合等离子体发射法,测定了我国4个主产区40余份当归药材及其土壤中14种无机元素的含量。结果:当归药材与土壤中Ca、Na、Ni显著相关;当归可能对Mg元素具有富集作用;Zn、Cu、Mn、Mg等元素可能是影响当归地道性最为显著的无机元素。实验结果从无机元素的角度支持岷归为道地药材。结论:当归药材所含无机元素类成分可能与其地道性呈现一定的相关性,为从认识当归的地道性成因及其药材功效物质基础提供参考。

炮制对当归药材有效成分的影响

目的建立当归药材及其炮制品的高效液相色谱测定法,比较当归炮制前后有效成分的变化。方法利用HPLC-DAD方法,梯度洗脱,测定了生当归及其炮制品中藁本内酯、阿魏酸的含量。色谱条件为:色谱柱Zorbax SB-C18柱(4.6mm×250mm,5μm);柱温25℃,以乙腈-0.01%磷酸水溶液为流动相,进行梯度洗脱,分析时间为60min。流速为0.8mL·min-1,检测波长220nm。结果当归经不同方法炮制后,其所含成分发生了变化。实验证明,该方法简便、可靠、重现性好。结论本方法可用于当归药材及其炮制品中藁本内酯及阿魏酸的含量测定,并为其全面质量控制提供参考;为探讨当归炮制意义,提供了一定的实验依据。

当归多糖AP-0对小鼠移植性肿瘤的抑制作用

目的 观察当归总多糖AP 0对于 3种小鼠移植性肿瘤的抑制作用。方法 小鼠肉瘤S180模型建立 :二级昆明种小鼠 ,每只鼠鼷部接种 5× 10 6瘤细胞。接种次日起 ,每日 1次定时ipAP 0 30mg·kg-1,10 0mg·kg-1和 30 0mg·kg-1,阳性对照组ip环磷酰胺 (Cy) 30mg·kg-1,共 10d ,第 11天活杀小鼠 ,称瘤质量、体质量、胸腺和脾质量 ,统计结果。小鼠Ehrlich腹水癌 (EAC)模型建立 :二级昆明种小鼠 ,每鼠ip接种 2 .5× 10 6瘤细胞。接种后次日起 ,每日 1次定时ipAP 0 30mg·kg-1,10 0mg·kg-1和 30 0mg·kg-1,阳性对照组ip氟尿嘧啶 ( 5 FU) 2 0mg·kg-1,共 10d ,观察各组 6 0d平均存活天数 ,计算生命延长率。小鼠白血病L12 10模型建立 :二级DBA/2小鼠 ,每鼠ip接种 1× 10 5瘤细胞。接种后次日起 ,每日 1次定时ipAP 0 30mg·kg-1,10 0mg·kg-1和 30 0mg·kg-1,阳性对照组ip环磷酰胺 (Cy) 30mg·kg-1,共 10d ;第 30天处死 ,计算生命延长率。结果 小鼠肉瘤 180实验结果 :瘤质量指标 :NS组和各治疗组瘤质量均大于Cy组 (P <0 .0 1) ;胸腺质量指标 :10 0mg·kg-1组胸腺质量小于NS组 (P <0 .0 5 ) ,30mg·kg-1和 30 0mg·kg-1组胸腺质量明显小于NS组 (P <0 .0 1) ,但 30mg·kg-1组胸腺质量大于Cy组 (P <0 .0 5 ) ,10 0mg·

当归不同药用部位水煎液指纹图谱及成分分析

目的建立当归Angelica sinensis(Oliv.)Diels不同药用部位水煎液HPLC指纹图谱,并分析其化学成分。方法全当归、当归头、当归身、当归尾水煎液的分析采用Ultimate XB-C18色谱柱(4.6 mm×250 mm,5μm);以0.1%甲酸-乙腈为流动相,梯度洗脱;柱温25℃;体积流量1 m L/min;检测波长280 nm。采用HPLC-MS联用技术分析其所含化学成分。结果以阿魏酸为参比峰,标定了24个特征峰,聚类分析表明全归和归身聚为一类,归头和归尾为一类。定性鉴定出8种化学成分,分别为色氨酸、绿原酸、阿魏酸、洋川芎内酯I、洋川芎内酯H、丁基苯酞、E-藁本内酯、Z-藁本内酯。结论该方法准确稳定,重复性好,可用于当归的质量控制。

当归HPLC指纹图谱研究(Ⅰ)

目的 建立甘肃当归药材HPLC指纹图谱。方法 以联苯为参照物 ,采用C18(2 5 0mm× 4 6mm ,5 μm)色谱柱 ,以甲醇 水 (水中含体积比为 0 5 %醋酸 )为流动相 ,梯度洗脱 ,体积流量 0 6mL/min ,检测波长为 2 75nm ,参比波长为 4 0 0nm。结果 精密度、重现性、稳定性试验中共有峰相对峰面积、相对保留时间的RSD均小于 5 0 % ;不同产地当归相似度大于 98 5 %。结论 该方法简便、实用、可靠 。

当归中有效成分阿魏酸的生物合成及调控

作为我国传统中药材当归的有效成分之一,含阿魏酸量的高低是决定当归品质的重要指标。现代分子生物学技术的发展为通过基因改造的方法提高当归品质提供了可能,这需要对阿魏酸的生物合成途径有详细的了解。综述阿魏酸在植物体内的生物合成机制及生物调控研究现状,为提高当归品质提供新的研究思路。

阿魏酸的稳定性及其在川芎和当归药材中的存在形式

目的 确认阿魏酸的异构体转换反应及川芎、当归药材中阿魏酸的存在形式。方法 采用高效液相色谱法、液相色谱 -质谱联用等手段研究阿魏酸的异构体转换。结果 反式阿魏酸在常规保存条件下 ,可部分转变成为顺式 ,在川芎和当归药材中 ,反式和顺式阿魏酸是共存的。结论 反式阿魏酸有较好的热稳定性 ,但存在缓慢的异构体转换反应。

以人血清白蛋白为固定相的分子生物色谱分析几种中药活性成分的研究

提出用人血清白蛋白分子生物色谱分析中药活性成分的方法 ,分别比较了当归等 4种单味中药的分离结果 ,考察了提取当归的 3种溶剂及提取时间对活性物质提取量的影响 ,同时建立了当归中阿魏酸与藁本内酯的定量分析方法 .

当归化学成分的分离与鉴定

目的研究当归Angelica sinensis(Oliv.)Diels的化学成分,为其药理作用的研究奠定物质基础。方法采用硅胶柱色谱及制备薄层色谱等方法进行分离纯化,根据理化性质和波谱数据(1H-NMR、13C-NMR等)鉴定化合物的结构。结果分离得到13个化合物,分别鉴定为:正丁基苯酞(3-butylphthalide,1)、4羟基3丁基苯酞(4-hydroxy-3-butylphthalide,2)、2-methoxy-2-(4'-hydroxyphenyl)ethanol(3)、反式阿魏酸(ferulic acid,4)、厚朴酚(magnolol,5)、黄芩苷(baicalin,6)、2″O(2甲基丁酰基)异当药黄素[2″O(2-methylbutyryl)-isoswertisin,7]、harman(8)、1-ace-tyl-β-carboline(9)、酒渣碱(flazine,10)、菠菜甾醇(α-spinasterol,11)、异欧前胡素(isoimperatorin,12)、发卡二醇[3(R),8(S)-falcarindiol,13]。结论化合物3、7～9、11为首次从该属植物中分离得到,化合物5为首次从该植物中分离得到。