Content Determination of Astragaloside Ⅳ in Astragalus from Three Different Regions by HPLC

[Objectives] The content of astragaloside in Astragalus membranaceus(Fisch.) Bge.var.mongholicus(Bge.) Hisao from three different regions was determined.[Methods] Referring to the method recorded in the Chinese Pharmacopoeia(2015 edition),the content of astragaloside IV in A.membranaceus was determined by HPLC.[Results] There were great differences in the astragaloside IV content of A.membranaceus among different regions.The content of astragaloside IV in A.membranaceus cultivated in Inner Mongolia was highest(0.155%),followed by that(0.143%) in A.membranaceus cultivated in Gansu,and the content of astragaloside IV in A.membranaceus cultivated in Shanxi was lowest(0.080%).The contents of astragaloside IV in A.membranaceus from different regions were all in line with the standard(not less than 0.040%) of Chinese Pharmacopoeia(2015 edition).[Conclusions]The content of astragaloside IV in A.membranaceus cultivated in three different regions met the medicinal standards.

Astragalus membranaceus(Fisch.)Bge.administered by dissolving microneedles achieves systemic therapeutic effects at low doses Author links open overlay panel

Objective:To determine the main components of Astragalus membranaceus(Fisch.)Bge(A.membranaceus,Huang Qi),Astragaloside IV(AIV)and Astragalus polysaccharides(AP),to characterize their properties,evaluate their in vivo efficacy,and to analyze drug diffusion using dissolving microneedle(DMN)technology in vivo.Methods: Respectively,AIV-and AP-loaded DMNs comprising chitosan(CTS)and polyvinyl alcohol(PVA)were prepared via dual-mold forming.Their morphology,mechanical properties,in vivo solubility,and skin irritation characteristics were tested.In vivo efficacy was assessed in cyclophosphamide-induced immunosuppressed mice,in vivo diffusion of AIV and AP by DMNs and conventional methods was compared,and the rheological properties of AIV-CTS-PVA and AP-CTS-PVA mixtures were measured.Results: Subcutaneous dissolution and absorption of AIV-CTS-PVA and AP-CTS-PVA microneedles(MNs)at low doses(50%–17%of intraperitoneal AIV injection and 12%–4%of intravenous AP injection)reduced the spleen index and acid phosphatase activity in immunosuppressed mouse models,increased the thymus index,and achieved equivalent or better systemic therapeutic effects.Compared with injections,AIV and AP achieved controllable solid-liquid conversion through delivery with CTS-PVA MNs,resulting in highly localized aggregation within 48 h,reducing the initial explosive effect of the drug,and achieving stable and slow drug release.Conclusion: The present study enhances our understanding of the efficacy and remote effects of drug-loaded DMNs from a traditional Chinese medicine(TCM)perspective,thereby promoting the development of precise and efficient delivery of TCM and further expanding the drug-loading range and application scenarios for DMNs.

Studies on Biologically Active Principles of Huangqi,Root of Astragalus membranaceous,Isolation and Detection of Constituents Scavenging Superoxide Anion

本文使用黄嘌呤-黄嘌呤氧化酶—鲁米诺化学发光体系和化学发光检测法研究了黄芪中各成分的清除超氧阴离子自由基的能力,以药物抑制发光强度50%的浓度(IC_(50))为指标。经研究证明,黄芪总皂甙部分(N)具有较强的活性,IC_(50)为185μg/ml,再经进一步导向分离并鉴定证明,黄芪甙Ⅲ,Ⅳ和Ⅵ的 IC)_(50)分别为80、50和11μg/ml,从而证明黄芪的抗心力衰竭的有效成分可能为黄芪总皂甙部分。

Molecular Mechanism of Induction on Apoptosis of Human Esophageal Cancer HCE-4 Cells by Active Components from Astragalus membranaceus

[Objectives] To investigate the pharmacologic effects of active components from A. membranaceus on human esophageal cancer HCE-4 cells and its apoptosis mechanism. [Methods] The viabilities of HCE-4 cells were measured by MTT assay. The induction of active components from A. membranaceus on apoptosis of HCE-4 cells was detected by Annexin V-FITC/PI double staining. The apoptotic-related protein expression levels were determined by Western blotting. [Results] Formononetin and astragaloside IV suppressed the proliferation of HCE-4 cells in a dose-dependent manner. The Annexin V-FITC/PI double staining results showed that formononetin and astragaloside IV could induce HCE-4 cells apoptosis in a time-dependent manner. The Western blotting results showed that formononetin and astragaloside IV could significantly down-regulate p-AKT,pro-caspase-3,and increase cle-caspase-3 protein expression in HCE-4 cells. [Conclusions]Active components from A. membranaceus such as formononetin and astragaloside IV significantly inhibited the proliferation of human esophageal cancer HCE-4 cells by inducing mitochondrial dependent apoptosis via AKT signaling pathway.

Functional characterization of a cycloartenol synthase and four glycosyltransferases in the biosynthesis of cycloastragenol-type astragalosides from Astragalus membranaceus

Astragalosides are the main active constituents of traditional Chinese medicine Huang-Qi,of which cycloastragenol-type glycosides are the most typical and major bioactive compounds.This kind of compounds exhibit various biological functions including cardiovascular protective,neuroprotective,etc.Owing to the limitations of natural sources and the difficulties encountered in chemical synthesis,re-engineering of biosynthetic machinery will offer an alternative and promising approach to producing astragalosides.However,the biosynthetic pathway for astragalosides remains elusive due to their complex structures and numerous reaction types and steps.Herein,guided by transcriptome and phylogenetic analyses,a cycloartenol synthase and four glycosyltransferases catalyzing the committed steps in the biosynthesis of such bioactive astragalosides were functionally characterized from Astragalus membranaceus.AmCAS1,the first reported cycloartenol synthase from Astragalus genus,is capable of catalyzing the formation of cycloartenol;AmUGT15,AmUGT14,AmUGT13,and AmUGT7 are four glycosyltransferases biochemically characterized to catalyze 3-O-xylosylation,3-O-glucosylation,25-O-glucosylation/O-xylosylation and 2’-O-glucosylation of cycloastragenol glycosides,respectively.These findings not only clarified the crucial enzymes for the biosynthesis and the molecular basis for the structural diversity of astragalosides in Astragalus plants,also paved the way for further completely deciphering the biosynthetic pathway and constructing an artificial pathway for their efficient production.

黄芪活性成分生理功能及在食品中的应用研究进展

黄芪作为最为常用的中药材,含有多种活性成分,其中最主要的有黄芪甲苷、黄芪黄酮和黄芪多糖。这些成分赋予黄芪提高机体免疫力、抗疲劳、消炎、保护心脑血管、保护内脏组织、抗癌抗肿瘤等多种药理作用,文章从黄芪活性成分的生理功能及在食品中的应用研究进展等方面进行综述,以期为黄芪及其活性成分在食品工业中的进一步开发应用提供理论参考。

基于CiteSpace和VOSviewer知识图谱分析黄芪抗炎功效的研究现状

目的利用CiteSpace和VOSviewer知识图谱对黄芪抗炎作用的研究现状进行可视化分析,为黄芪在抗炎方面研究提供参考。方法以“黄芪”和“抗炎”为主题检索词,分别从中国知网数据库(CNKI)和Web of Science数据库检索中英文文献,利用CiteSpace、VOSviewer及Excel软件对中英文文献的发文量、作者、发文机构、被引频次、关键词等进行可视化分析。结果共纳入有效文献386篇,研究黄芪抗炎功效的文献年发文量总体呈增长趋势。中英文文献发文量居首位的机构分别是天津中医药大学和中国科学院;发文量最多的作者分别是张雪梅和Dean Guo(果德安)。关键词共现、聚类显示研究热点为黄芪甲苷、黄芪多糖抗炎的药理作用以及作用机制等。结论研究黄芪对炎症药理作用的文章逐年增加,且热点主要集中在黄芪甲苷和黄芪多糖。

黄芪活性成分治疗呼吸道感染药理作用的研究进展

黄芪活性成分具有良好抗菌活性,抑制病原菌生长,还能增强疫苗抗病毒活性,以防治呼吸道感染。黄芪活性成分通过下调Toll样受体(Toll-like receptors,TLR)4/丝裂原活化蛋白激酶(mitogen-activated protein kinase,MAPK)/核因子κB(nuclear factor kappa-B,NF-κB)、TLR4/7/髓样分化初级反应蛋白88(myeloid differentiation primary response protein 88,MyD88)/NF-κB信号通路,抑制NOD样受体蛋白3(NOD-like receptor protein 3,NLRP3)炎症小体活化,刺激M2巨噬细胞极化,抑制转化生长因子β1(transforming growth factor-β1,TGF-β1)/Smad信号通路,增强自噬,减轻炎症反应,以减轻呼吸道感染引起的炎性损伤。黄芪活性成分通过调节氧化应激产物的分泌,调控TLR3、血红素氧合酶1(heme oxygenase-1,HO-1)/核因子E2相关因子-2(nuclear factor E2 related factor-2,Nrf2)信号通路,减轻氧化应激反应,以减轻肺部感染引起的氧化应激损伤。黄芪活性成分调节T淋巴细胞、辅助性T淋巴细胞17(T-helper 17,Th17)/调节性T淋巴细胞(regulatory T lymphocyte,Treg)平衡,增强疫苗免疫反应,调节机体免疫功能,增强对病原菌的抵抗力。黄芪活性成分通过调控凋亡相关蛋白的表达,调控激活沉默信息调节因子1(silent information regulator1,SIRT1)/腺苷酸活化蛋白激酶(adenylate activates protein kinase,AMPK)信号通路,激活磷脂酰肌醇-3激酶(phosphatidylinositol-3 kinase,PI3K)/蛋白激酶B(protein kinase B,Akt)信号通路,减轻细胞凋亡,对肺组织发挥保护作用。

不同产地黄芪饮片中多糖及甲苷含量比较

黄芪是常用补气药之一,黄芪多糖和黄芪甲苷是其发挥药效的物质基础。以黄芪多糖和黄芪甲苷为评价指标,比较甘肃、内蒙古、山西三个不同产地的黄芪饮片的质量差异。结果表明:甘肃黄芪、内蒙古黄芪、山西黄芪的多糖含量分别为22.87%、20.02%、22.29%;甘肃黄芪的黄芪甲苷含量最高,为0.049%;其次是山西黄芪,黄芪甲苷含量为0.026%,内蒙古黄芪中黄芪甲苷含量最低,仅为0.0027%。仅有甘肃黄芪的甲苷含量测定结果符合药典标准。分析认为药材生长年限是导致内蒙古黄芪与甘肃黄芪、山西黄芪甲苷含量之间差异较大的主要原因。该研究为黄芪种植、资源开发及合理使用提供参考依据。

基于主成分分析法研究干燥方法对黄芪药材质量的影响

目的:研究五种产地加工干燥方法对黄芪药材的外观性状、内在成分的影响规律,以期选出最佳的黄芪药材产地初加工干燥方法。方法:对新鲜黄芪药材进行晒干、阴干、热风干燥(50、60、70℃)的处理,比较干燥速率随干燥时间的动态变化规律,研究其对A药材外观性状、浸出物、复水比;采用HPLC-ELSD测定黄芪甲苷含量;HPLC-UV法测定毛蕊异黄酮葡萄糖苷含量;UV-Vis测定黄芪多糖含量,最后采用主成分分析法进行综合评价。结果:经对实验数据分析,黄芪物效含量为60℃烘干>晒干>阴干>50℃烘干=70℃烘干,其中60℃烘干和晒干的贡献得率达93.936%,具有绝对优势。阴干,晒干,50℃、60℃、70℃热风干燥五种干燥方法使黄芪达到安全含水(10%)所需时间分别为132 h、60 h、13.5 h、9.5 h、4.5 h。结论:从黄芪外观性状与内在成分含量等因素考虑,其产地加工方法以60℃热风干燥为宜,但综合经济因素考虑,晒干亦较为适宜。

黄芪苷 Ⅳ对正常和心功能受抑制大鼠左心室心肌力学的影响

目的 研究黄芪苷 对正常和心功能受抑制大鼠左心室心肌力学的效应。方法 采用左心室导管法 ,测定左心室压力 (L VP)及其微分 (dp/dt) ,对 iv黄芪苷 (1m g/kg)对左心室心肌力学的效应。iv普萘洛尔 (0 .75m g/kg)抑制大鼠心功能 ,观察黄芪苷 的对抗作用。结果 黄芪苷 使心率 (HR)略有减慢 ;使左室压力上升最大速度 (dp/dtmax)明显提高 ,具有正性肌力作用 ;使左室压力下降的时间常数 (T)显著缩短 ,改善心脏舒张功能 ;左心室射血的张力 -时间指数 (TTI)无明显改变 ,表明该药在增强心肌收缩力的同时并未增加心肌耗氧量。对普萘洛尔抑制心功能的大鼠 ,用黄芪苷 后 dp/dtmax无显著降低 ,即该药可对抗普萘洛尔抑制心肌收缩力的作用 ;T值显著缩短 ,表明心脏舒张功能也有改善 ;心肌耗氧量不增加。结论 黄芪苷 可改善心脏收缩及舒张功能 。

黄芪和当归的主要活性成分配伍对骨髓抑制小鼠造血功能的影响

目的探讨黄芪和当归的主要活性成分配伍对骨髓抑制小鼠模型的促造血作用。方法对芪归1∶1配伍提取物测定5种主要活性成分阿魏酸、芒柄花素、黄芪甲苷、毛蕊异黄酮、毛蕊异黄酮苷含量,以确定配伍剂量。小鼠随机分为空白对照组、模型组和各给药组。空白对照组给予溶剂灌胃7 d;模型组同上灌胃,于d 3腹腔注射环磷酰胺,连续3 d;各药物组灌胃给药,同上造模。检测外周血象、血清造血生长因子(HGF)含量、造血祖细胞集落数。结果芪归配伍提取物和活性成分配伍均可明显增加外周血白细胞(WBC)、红细胞(RBC)、血小板数(Pt)、血红蛋白(HGB)含量和骨髓造血组织面积,明显增加血清粒-巨噬细胞集落刺激因子(GM-CSF)、促血小板生成素(TPO)、促红细胞生成素(EPO)含量和骨髓粒-单核细胞集落形成单位(CFU-GM)、巨核细胞集落形成单位(CFU-MK)、红系集落形成单位(CFU-E)和爆式红系集落形成单位(BFU-E)数,两者作用相近。各成分中,阿魏酸可促进WBC和骨髓造血组织面积的恢复;5种成分均可促进RBC和HGB的恢复;阿魏酸和黄芪甲苷可促进Pt的恢复。5个成分均可升高TPO含量;阿魏酸和毛蕊异黄酮可升高GM-CSF含量;阿魏酸、毛蕊异黄酮和毛蕊异黄酮苷可升高EPO含量。除黄芪甲苷外,其它4种成分均可使CFU-GM增加;5种活性成分均可增加CFU-MK、CFU-E;仅阿魏酸可增加BFU-E。结论黄芪当归配伍发挥补血作用的主要药效物质是以上5种活性成分,这些活性成分配伍对促进造血可发挥增效作用。

安国引种黄芪的质量初步评介

测定安国县１０个村引种黄芪药材中黄芪甲苷含量，平均值为０．１９９％，比传统道地黄芪高２倍，比《中国药典》标准（不得少于０．０４％）高近４倍，但外观质量难以相比，１０个村黄芪药材中砷、汞、铅、镉、镍、钴等 ６种元素的平均值分别为 ０． ２４４、０． ００９、１．７３１、０． ０３６、０． ２１４、０． ３０２ μｇ／ｇ，与土壤中６种元素的比值即富集系数≤０．５。

黄芪皂苷和黄芪多糖对大鼠肾脏系膜细胞增殖的影响

目的观察黄芪皂苷和黄芪多糖对体外培养的肾脏系膜细胞增殖的影响。方法分离和培养大鼠肾脏系膜细胞,用MTT法研究不同浓度的皂苷和多糖对细胞增殖的影响,并计算抑制率。结果黄芪多糖在低浓度时表现出一定的促进细胞增殖作用,在高浓度时有一定的抑制作用;黄芪皂苷在所考察的范围内均表现出一定的抑制作用。结论黄芪皂苷和黄芪多糖在一定浓度时对肾脏系膜细胞增殖具有一定的抑制作用,为其在治疗肾小球肾炎中的作用提供了理论依据。

HPLC-ELSD法测定黄芪中黄芪甲苷的含量

目的 采用HPLC．ELSD法测定黄芪中黄芪甲苷的含量。方法 HPLC-ELSD法，汉邦C18柱（4．6mm×150mm），流动相：乙腈-水（40：60），流速：1．0ml；ELSD参数：漂移管温度：105℃，载气流速为2．8L·min^-1。结果黄芪甲苷在4．0～18．0μg范围内有良好的线性关系，r=0．9999（n=6），平均回收率为98．0％。结论 该方法分离度高，重现性好，简便，准确，可广泛用于黄芪药材的质量控制。研究发现，实验所用样品前处理过程中是否用氨试液洗涤，显著影响黄蓖甲苷的含量测定。

黄芪甲苷在兔体内的药动学和在大鼠的排泄(英文)

目的 :研究黄芪甲苷在家兔体内的药动学和在大鼠的排泄。方法 :健康家兔和大鼠一次静脉注射 (静注 )给予黄芪甲苷 4mg·kg- 1,高效液相色谱 蒸发光散射检测器法检测兔血浆和大鼠尿及粪黄芪甲苷浓度 ,用 3P97药动学软件对兔血浆浓度 时间数据进行动力学分析和计算药动学参数 ,并估算大鼠体内的排泄情况。结果 :黄芪甲苷静注给药后 ,T12 α 为 0 .10h ,T12 β 为 1.4h ,Vc 为 0 .15L·kg- 1,VD 为 0 .6L·kg- 1,Cl为 0 .32L·h- 1·kg- 1,AUC为 15mg·L- 1·h。大鼠静注给药后 ,原形从尿和粪排出量分别为给药量的 16 %和 3.2 %。结论 :家兔体内黄芪甲苷的动力学过程符合二室模型 ,大鼠仅有少量原形药物从尿和粪排泄。

黄芪甲苷、总多糖、注射液对3种人恶性肿瘤细胞增殖的影响

目的:比较黄芪不同制剂、成分对3种人肿瘤细胞株SMMC7721、PC3、HL60增殖的影响。方法:将不同浓度黄芪甲苷、总多糖、注射液,直接加入肿瘤细胞培养液中,MTT法测定各成分对三种肿瘤细胞的增殖影响。结果:黄芪甲苷、总多糖、注射液上述三种人肿瘤细胞增殖无明显抑制作用,且黄芪甲苷在20mg/ml浓度时对SMMC7721、PC3的增殖有一定促进作用。结论:体外实验结果表明黄芪甲苷、总多糖、注射液对三种人肿瘤细胞株增殖无明显抑制作用。

黄芪多糖及黄芪甲苷对巨噬细胞吞噬结核杆菌作用的研究

目的:探讨黄芪多糖或黄芪甲苷对巨噬细胞吞噬结核杆菌作用的影响。方法:将不同浓度黄芪多糖或黄芪甲苷小鼠腹腔巨噬细胞及结核杆菌共同孵育,检测巨噬细胞对结核杆菌的吞噬能力,并检测培养液中IFN-γ和IL-1β的含量。结果:当黄芪多糖或黄芪甲苷的给药量分别为0.2、0.6、1.5、4μg/μl时,巨噬细胞吞噬结核杆菌的TB-DNA拷贝数以及上清液中IFN-γ和IL-1β的含量均明显高于对照组。结论:黄芪多糖或黄芪甲苷均提高巨噬细胞对结核杆菌的吞噬作用,并以黄芪甲苷效果更佳,但它们均具有双相性。

发酵对黄芪中黄芪甲苷含量的影响

建立HPLC-UV法检测黄芪甲苷含量,比较固态发酵对发酵黄芪中黄芪甲苷含量的影响。采用色谱条件:Inertsustain C18色谱柱(150mm×4.6mm,5μm),流动相为乙腈-水(32∶68),检测波长203nm,柱温25℃,流速1mL/min,进样量20μL。结果表明,该方法在0.06mg/mL^0.48mg/mL范围内线性关系良好(R^2=0.999 3)。经检测,发酵黄芪中黄芪甲苷的含量为6.336mg/g,非发酵黄芪中黄芪甲苷含量为4.593 mg/g,发酵黄芪较对照黄芪含量增加了37.95%,黄芪经发酵后可使黄芪甲苷含量明显升高。

LC-MS-MS法同时测定黄芪注射液和黄芪口服液中5种成分

目的建立LC-MS-MS法同时测定黄芪注射液和黄芪口服液中黄芪甲苷、毛蕊异黄酮葡萄糖苷、芒柄花苷、毛蕊异黄酮、芒柄花黄素5种有效成分的含有量。方法黄芪注射液和黄芪口服液甲醇稀释液在Agilent ZORBAX SB C18（2.1 mm×150 mm,5μm）色谱柱上分离,流动相为甲醇-水（含0.1%甲酸,70∶30,V/V）;体积流量为300μL/min。MS采用电喷雾离子源,多反应监测方式进行正离子扫描。结果 5种成分在测定浓度范围内均具有良好的线性关系,在黄芪注射液和黄芪口服液中的平均回收率分别为96.8%-102.3%和95.9%-101.5%。结论本法测得结果准确、可靠、灵敏度高,可用于黄芪注射液和黄芪口服液的质量控制。