Structural characterization and mechanisms of macrophage immunomodulatory activity of a novel polysaccharide with a galactose backbone from the processed Polygonati Rhizoma

A purified polysaccharide with a galactose backbone(SPR-1,Mw 3,622 Da)was isolated from processed Polygonati Rhizoma with black beans(PRWB)and characterized its chemical properties.The backbone of SPR-1 consisted of[(4)-b-D-Galp-(1]9/4,6)-b-D-Galp-(1/4)-a-D-GalpA-(1/4)-a-D-GalpA-(1/4)-aD-Glcp-(1/4,6)-a-D-Glcp-(1/4)-a/b-D-Glcp,with a branch chain of R1:b-D-Galp-(1/3)-b-D-Galp-(1/connected to the/4,6)-b-D-Galp-(1/via O-6,and a branch chain of R2:a-D-Glcp-(1/6)-a-D-Glcp-(1/connected to the/4,6)-a-D-Glcp-(1/via O-6.Immunomodulatory assays showed that the SPR-1 significantly activated macrophages,and increased secretion of NO and cytokines(i.e.,IL-1b and TNF-a),as well as promoted the phagocytic activities of cells.Furthermore,isothermal titration calorimetry(ITC)analysis and molecular docking results indicated high-affinity binding between SPR-1 and MD2 with the equilibrium dissociation constant(KD)of 18.8 mM.It was suggested that SPR-1 activated the immune response through Toll-like receptor 4(TLR4)signaling and downstream responses.Our research demonstrated that the SPR-1 has a promising candidate from PRWB for the TLR4 agonist to induce immune response,and also provided an easily accessible way that can be used for PR deep processing。

Fingerprint Study of Polygonati Rhizoma with Steaming and Exposing to the Sun Alternatively for Different Times

[Objectives]To explore the influence of different times of steaming and exposing to the sun on the fingerprint of Polygonati Rhizoma by studying the HPLC fingerprint of Polygonati Rhizoma processed products with different times of steaming and exposing to the sun,and to provide a basis for the determination of the best processing technology of Polygonati Rhizoma.[Methods]SETSAIL II AQ-C 18(5μm×250 mm×4.6 mm)was used as the column,the column temperature was 30℃,pure water(A)and acetonitrile(B)were eluted gradually,0-10 min,B(5%-10%),10-30 min,B(10%-35%),30-40 min,B(35%-60%),40-45 min,B(60%-100%),flow rate 1 mL/min,absorption wavelength 200 nm.[Results]The relative retained peak area RSDs of the common peaks in the precision,reproducibility and stability tests were all less than 5%.There were 17 common peaks in the fingerprint of nine batches of samples,and the retention time of Peak 2 was basically the same as that of the reference peak of 5-HMF.Peak 4 mainly existed in the chromatogram of Sample 3 to Sample 5,peaks 5 and 11 mainly existed after Sample 3,peaks 9,14 and 16 mainly existed after Sample 6,and peaks 12 and 17 mainly existed after Sample 4.[Conclusions]A total of 17 common peaks were obtained,and the Peak 2 was the designated peak,and the chemical components of each processed product were different.

The Inhibitory Effect of Extracts From Fructus lycii and Rhizoma polygonati on in vitro DNA Breakage by Alternariol

Alternariol caused DNA single-strand breakage. Conversion of the closed circular double-stranded supercoiled DNA (pBR 322) to the nicked circular form and linear form was used to investigate the effect of extracts of some Chinese medical herbs on DNA nicking induced by alternariol. Some substances in the extracts of Rhizoma polygonati (RP) and Fructus lycii (FL) were shown to protect DNA from the attack by alternariol.Some substance in the RP may bind to plasmid DNA, and this binding reduces the electrophoretic mobility of DNA. These results indicate that substances from FL and RP may be used as DNA protectors. It is possible that they play an important role in preventing cancer.

Botany,Traditional Uses,and Pharmacology of Polygonati Rhizoma

Huang Jing (黄精 Polygonati Rhizoma, PR) was first documented as a herbal medicine in Ming Yi Bie Lu (《名医别录》 Miscellaneous Records of Famous Physicians) in China. However, there was no comprehensive review on the botany, traditional uses, and pharmacological effects of PR till now. In this study, the botany, traditional uses including Taoist medicine, and pharmacological effects of PR were reviewed and summarized to provide insights on drug development of PR. In Taoist medicine, PR maintains agerasia and helps prolong human life-span, and is used for fasting (Bigu). In the Zhong Guo Yao Dian (《中国药典》Chinese Pharmacopeia) version 2020, PR exerts replenishing qi and nourishing yin, invigorating the spleen, moistening the lung, and strengthening the kidney. Pharmacological studies show that PR has effects of anti-oxidation, anti-diabetes, anti-osteoporosis, anti-cancer, anti-hyperlipidemia, cardiomyocyte protection, immunomodulatory, and thus can be used for treatment of infertility, anti-microorganisms, and improving sleep and memory. In conclusion, PR may play a potential role for chronic disease management and health preservation and this very role deserves a more in-depth research.

Extraction Optimization of Polysaccharides from Rhizoma Polygonati by Response Surface Methodology and Content Determination

[Objectives]This study aimed to investigate the water extraction-alcohol precipitation technology and content determination method of polysaccharides in Rhizoma Polygonati.[Methods]Taking the content of polysaccharides in the extract as the indicator,the extraction process was optimized using single-factor tests and Box-Behnken Design.The content of polysaccharides extracted was determined by the anthrone-sulfuric acid method.Among the three factors in the test design,the solid/liquid ratio and extraction time had a significant effect(P<0.05,P<0.01),while the extraction temperature had no effect on the content of polysaccharides extracted(P>0.05).[Results]The optimal extraction process for polysaccharides in Rhizoma Polygonati was as follows:liquid/solid ratio 20∶1(mL/g),extraction time 2 h,and extraction temperature 85℃.Under the optimal extraction process,the content of polysaccharides in the extract reached 12.50%.[Conclusions]The extraction process optimized by response surface methodology for polysaccharides in Rhizoma Polygonati is reliable and has practical value,and can be used to guide production practices.

Rhizoma polygonati from Mount Tai:nutritional value and usefulness as a traditional Chinese medicine,source of herbzyme,and potential remediating agent for COVID-19 and chronic and hidden hunger

Recently,traditional Chinese medicine-based treatment has succeeded in fighting coronavirus disease 2019(COVID-19),and Rhizoma polygonati(Huangjing)has been one of the recommended components.Its processed products play antidiabetic,antiviral,antitumor,antioxidation,antifatigue,antiaging,and immune enhancement roles.The climate in Mount Tai is mild,and the dense forest is suitable for the growth of Rhizome polygonati,which has gradually evolved into a unique specie.Considering the important medicinal value and pleasant taste of Mount Tai-Rhizoma polygonati,various healthy and functional food products,controlled by quality markers with anti-COVID-19 potential,as well as emergency foods can be developed.The study aimed to review current evidence on the nutritional value of Rhizoma polygonati from Mount Tai and its usefulness as a traditional Chinese medicine,source of herbzyme,and potential remediating agent for COVID-19 and food shortage.Most recent findings regarding herbal nanomedicine have revealed that nanoscale chemical compounds are potentially efficient in drug delivery or nanozyme catalysis upon bioprocessing.Nanoflower structure is found in processed Rhizoma polygonati by self-assembly and has wide application in enzymatic events,particularly nanoscale herbzyme.The novel findings regarding Mount Tai-Rhizoma polygonati could enhance its novel applications in chronic and hidden hunger,clinical nanomedicine,and as an anti-COVID-19 agent.

Qualitative and Quantitative Analysis of Linoleic Acid in Polygonati Rhizoma

Objective To explore the major compound in Polygonati Rhizoma(Huang Jing,黄精)for quality control.Methods The major compound was isolated and analyzed by liquid chromatography-mass spectrometry(LC-MS),and subsequently further identified by nuclear magnetic resonance(NMR).Thin layer chromatography(TLC)was optimized based on the previous methods reported in the Chinese Pharmacopeia(2015 edition).Results The major compound was isolated from the natural material and identified as linoleic acid.A high performance liquid chromatography(HPLC)method with robust linearity(R2=0.9997),specificity,precision,stability,repeatability and recovery was developed for linoleic acid determination.TLC chromatogram was improved significantly after optimization for qualitative analysis.Conclusions The optimized TLC method is practical and can be adopted for quality control of Polygonati Rhizoma(Huang Jing,黄精).The levels of linoleic acid vary between species of Polygonati Rhizoma(Huang Jing,黄精),with Polygonatum cyrtonema Hua(Jiang Xing Huang Jing,姜型黄精)showing the highest contents.This study provides valuable information for quality control of Polygonati Rhizoma(Huang Jing,黄精).

Characterization of metabolite landscape distinguishes wild from cultivated Polygonati Rhizomes by UHPLC-Q-TOF-MS untargeted metabolomics

Rhizome,a modified subterranean plant stem,has been used in China for centuries as a Traditional Chinese Medicine(TCM).Polygonati Rhizome(PR)is a well-known species in the Rhizome family.Previous studies show that PR might be beneficial in the treatment of osteoporosis,cancer,diabetes,and many other diseases because of its properties such as antioxidant property.Thus cultivated PR strains were developed to fulfil the increasing demands.However,the metabolite profile differences between wild-type and cultivated PR remain largely unknown.Here,we performed unbiased and untargeted quantitative mass spectrometry-based metabolomics on 1 wild strain and 3 cultivated strains.A total of 1126 metabolites were identified on all 4 strains.Analyses of these data with unsupervised and supervised approaches identified common metabolites in all 4 strains as well as strain-specific metabolites.Next,we conducted a metabolomic-wide pairwise correlation analysis on metabolite abundance and discovered 577 significantly correlated metabolite-metabolite pairs by Pearson’s correlation test.Importantly,KEGG enrichment analysis indicated that phenylpropanoids biosynthesis was the most differentially expressed pathway between cultivated and wild-type PR.By contrast,common metabolites on all 4 strains might suggest shared therapeutic targets.In summary,the systematic,untargeted metabolomics profiling of 4 PR strains and comprehensive data analyses provide invaluable resources to the understanding of the metabolite landscape and shed light on potential therapeutic applications for the treatment of diseases.

黄精多糖提取工艺优化及其吸湿、保湿性能研究

目的:优化黄精多糖提取工艺,并研究其吸湿保湿性能、探索其护肤功能。方法:采用热回流法结合正交进行黄精多糖提取工艺优化;以黄精多糖为实验对象,采用重量法进行吸湿性(相对湿度分别为43%和81%)、保湿性试验,并对吸湿曲线进行模型拟合;将黄精多糖添加到市售面霜中进行皮肤水分测试。结果:实验表明黄精多糖最佳提取工艺条件为:料液比1∶25(g/mL),提取时间是150 min,提取次数为1次;黄精多糖在不同相对湿度下的吸湿均符合双指数模型;置于干硅胶环境下48 h,黄精多糖保湿率为14.00%。结论:黄精多糖具有良好的保湿性,在护肤品开发中具有一定的应用价值。

玉竹的化学成分、药理作用及其食品开发研究进展

玉竹不仅富含糖、蛋白质、氨基酸、维生素和矿物质等营养成分,还含较高的多糖、甾体皂苷、黄酮等生物活性成分,具有抗氧化、调节糖脂代谢、增强免疫、抗肿瘤、抗微生物、抗疲劳、抗衰老、护肝、调节胃肠道菌群、减肥等药理作用,已广泛用于普通食品和健康食品的开发利用。本文对其营养功能成分、生物活性及其在食品加工领域中的应用进展进行系统总结,以为其深入开发提供参考和支持。

基于质量源于设计(QbD)理念优化黄精粗多糖制备工艺及抗氧化活性检测

试验旨在优化复合酶协同超声水提醇沉法制备黄精粗多糖(crude polysaccharide of po⁃lygahatous,CPP)的工艺并检测其体外抗氧化活性。试验以质量源于设计(qulity by design,QbD)理念为指导,以黄精多糖得率和含量为指标,应用Box-Behnken响应面设计法(BBD)对CPP制备工艺的关键因素:酶解温度、加酶量、超声时间、醇沉浓度进行优化;以维生素C(Vitamin C,VC)为对照,对CPP进行1,1-二苯基-2-三硝基苯肼(1,1-Diphenyl-2-picrylhydrazyl,DPPH)自由基和羟基(·OH)自由基清除能力的检测,评价CPP体外抗氧化活性。结果表明:CPP最佳制备工艺为酶解温度55℃,复合酶(纤维素酶∶果胶酶=1∶1)用量0.28%,超声时间50 min,醇沉浓度78%;采用该制备工艺,CPP的得率为(28.12±1.43)%,多糖含量为(33.74±1.32)%。不同浓度CPP能够有效清除DPPH自由基和抑制·OH的产生。由此可见,优化的CPP制备工艺稳定可行,且所制得的CPP具有一定的体外抗氧化活性。

黄精转录组SSR分子标记开发及种质遗传多样性分析

为丰富可用于黄精属鉴定的SSR分子标记,开发可用于黄精属(Polygonatum sp.)种质资源鉴定与遗传多样性分析的简单重复序列(simple sequence repeat,SSR)分子标记,为药材黄精正品和伪品的鉴定及遗传多样性分析提供基础。通过对3种黄精属植物进行转录组测序,从60836967个Unigenes鉴定出19630个包含1~6个碱基重复类型的SSR位点,设计和筛选获得了9对SSR有效性引物,并对来源于5个省份共计31份种质资源进行鉴定与遗传多样性分析。9对SSR引物不仅能够将长梗黄精与其他3种药材黄精划分开,还能够较好地区分不同基源黄精(滇黄精、黄精、多花黄精),并且31份资源遗传相似系数范围为0.0769~0.75。此外,构建了31份黄精属种质资源的DNA指纹图谱数据库,开发了相对应的指纹图谱。本研究结果为黄精属种质资源的分类鉴定、遗传多样性分析和品种选育提供了可用的分子标记以及参考依据。

基于响应面法的九制黄精炮制工艺优化

【目的】优化九制黄精炮制工艺,为九制黄精的质量控制提供技术参考。【方法】采用紫外-可见分光光度法,以黄精多糖含量为指标,以蒸制温度、蒸制时间、焖润时间、干燥温度及蒸制次数为影响因素设计单因素试验;依据单因素试验结果,以蒸制时间、焖润时间和蒸制次数为3因素,通过Box-Benhnken设计3因素3水平响应面优化试验。【结果】在单因素试验中,蒸制温度从101℃到109℃时,黄精多糖含量为3.14%~3.31%,蒸制温度105℃时,黄精多糖含量测定值最高,为3.31%,但不同蒸制温度之间在统计学意义上差异不显著(p>0.05);干燥温度从50℃升高到90℃过程中,黄精多糖的含量呈逐步下降趋势,其中蒸制温度50℃时黄精多糖含量最高。响应面法优化试验得到最佳的炮制工艺参数为蒸制时间为2 h、焖润时间为5.3 h、蒸制次数为6次,在此条件下得到黄精多糖含量为5.14%。【结论】九制黄精炮制工艺参数的优化,在技术上为九制黄精现代化、规模化生产提供重要参考依据。

高能电子束辐照对黄精微生物及品质的影响

为探究高能电子束辐照对黄精的影响,以未辐照样品为对照,研究不同剂量(2、4、6、8、10 kGy)电子束辐照对黄精微生物含量、水分、总灰分、浸出物及色泽、挥发性物质的影响。结果表明,经2 kGy剂量辐照,黄精中微生物总数降低至《中国药典》要求限度;随着剂量增加,辐照杀菌效果更好。辐照对黄精水分和总灰分含量无显著影响(P>0.05)。辐照剂量≥4 kGy时,黄精浸出物含量、色度黄蓝值(b^(*)值)和饱和度显著增加(P<0.05),但不影响总色泽。辐照对黄精主要挥发性成分无明显影响,但在一定剂量下,会加速芳香成分、氮氧化合物、甲烷物质及醇类、醛酮类化合物挥发;主成分分析及线性判别分析表明,2 kGy剂量辐照对黄精挥发性影响最小。综合分析,2 kGy剂量高能电子束辐照能有效杀灭黄精中的微生物,且对其水分、总灰分、浸出物、色泽及挥发性成分无显著影响。

高能电子束辐照对黄精品质的影响

以药食同源中药材黄精为原料,采用辐照剂量为0、2、4、6、8 kGy的高能电子束对其进行处理,分析不同辐照剂量下其微生物含量、主要活性物质含量、高效液相色谱(high performance liquid chromatography,HPLC)指纹图谱、色泽、水分、灰分和浸出物等的变化情况,探究高能电子束辐照对黄精品质的作用。结果显示,经高能电子束辐照后,能有效减少黄精中需氧菌、霉菌和酵母菌的总数;低辐照剂量后,黄精活性成分的含量无显著性变化(P>0.05);黄精HPLC指纹图谱相似度均大于0.90,表明经不同辐照剂量的电子束辐照处理后黄精的指纹图谱整体仍较为一致且稳定;当辐照剂量为2、4 kGy时,其色泽与对照组相比无显著性变化(P>0.05);各辐照剂量下黄精的水分、灰分和醇溶性浸出物均符合2020年版《中国药典》的相关规定。综上,当电子束的辐照剂量为2 kGy时,既能使黄精微生物数量明显降低,又能够保障其品质。

黄精药食同源价值研究进展

药食同源思想源远流长,体现了中医药特色。黄精作为药食同源传统中药材,对延缓衰老、预防和治疗疾病有显著效果,在医药、保健食品等领域开发前景广阔。本文综述了药食同源文化来源及现状,分析了黄精的药膳起源、化学成分、药理作用以及加工应用方面的最新进展,结合湖南黄精的种植情况,为黄精资源进一步开发提供参考依据。

黄精化学成分、药理作用研究进展及质量标志物预测分析

黄精是常用的补益药,主要炮制品有酒黄精、蒸黄精、蜜黄精。笔者查阅相关文献,对黄精的化学成分、药理作用进行总结。发现黄精的化学成分多样,主要为皂苷类、多糖类、氨基酸类、黄酮类等,经过炮制后其多糖类、皂苷类、氨基酸类、黄酮类、蒽醌类及矿物质成分含量发生明显变化;具有抗衰老、抗氧化、抗糖尿病、抗骨质疏松及免疫调节作用;从传统药性、炮制前后化学成分变化、化学成分可测性方面分析,初步确定黄精多糖、甾体皂苷类、氨基酸类、酚类、黄酮类可作为其质量标志物(Q-Marker)的参考。

基于网络药理学和分子对接探讨熟地黄—黄精治疗阿尔茨海默病的作用机制

目的 整合网络药理学、分子对接技术探究熟地黄—黄精治疗阿尔茨海默病(Alzheimer’s disease, AD)的作用机制。方法 在中药系统药理数据库(Traditional Chinese Medicine System Pharmacology, TCMSP)数据库中收集熟地黄、黄精化学成分和潜在作用靶点,在在线人类孟德尔遗传(online mendelian inheritance in man, OMIM)、治疗靶点(therapeutic target database, TTD)、GeneCards、Drug Bank数据库收集AD疾病靶点基因,运用Cytoscape3.9.1构建“中药—成分—靶点”网络;将熟地黄、黄精与AD的交集靶点基因上传至STRING数据库中分析核心靶点基因;将交集靶点上传至DAVID数据库中进行GO和KEGG富集分析,并通过分子对接检测核心化学成分和核心靶点基因的结合能力。结果 在熟地黄、黄精中共筛选出13个化学成分,其中薯蓣皂苷、3′-甲氧基黄豆苷、豆甾醇、黄芩苷、β-谷甾醇为核心化学成分;AD相关靶点共筛选出2471个,熟地黄、黄精与AD的交集靶点共有68个;PPI分析显示,蛋白激酶B(protein kinase B,Akt1)、前列腺素G/H合成酶2(prostaglandin G/H synthase 2,PTGS2)、凋亡相关蛋白Caspase-3等为核心交集靶点基因;GO分析显示,生物过程主要涉及对含氧化合物、对化学物质的反应等,细胞组成主要涉及神经元投射、神经元部分、躯体树突间室等,分子功能主要涉及酶结合、G蛋白偶联胺受体活性、相同蛋白质结合等;KEGG分析结果显示,核心靶点基因涉及的通路主要为内分泌抵抗、麻疹、雌激素信号通路、凋亡、流体剪切应力与动脉粥样硬化等;分子对接结果显示,核心化学成分黄芩苷、β-谷甾醇、3′-甲氧基黄豆苷与核心靶点结合效能高。结论 熟地黄、黄精防治AD的成分主要为黄芩苷、β-谷甾醇、3′-甲氧基黄豆苷等,这些成分可通过调节10个核心靶点基因发挥防治AD的作用,其机制可能与凋亡、雌激素通路等5个途径有关。

基于UPLC-Q-TOF-MS的黄精炮制前后低聚糖成分分析及抗氧化活性研究

目的:探究黄精酒制前后低聚糖的成分种类、变化情况,以及生、酒黄精低聚糖抗氧化活性。方法:采用超高效液相色谱串联四极杆飞行时间高分辨质谱对1-苯基-3-甲基-5-吡唑啉酮(PMP)衍生化后的低聚糖进行成分分析,采用苯酚-硫酸法测定低聚糖含量,采用1,1-二苯基-2-三硝基苯肼(DPPH)自由基清除法和2,2-联氮-二(3-乙基-苯并噻唑-6-磺酸)二铵盐(ABTS+)自由基清除法测定其体外抗氧化活性,利用秀丽隐杆线虫测定体内抗氧化活性。结果:在生、酒黄精低聚糖中同时检测到蔗糖、蔗果三糖和蔗果四糖3种低聚糖,蔗糖、蔗果三糖、蔗果四糖在炮制后减少,棉子糖、蜜二糖、蔗果五糖、蔗果六糖在炮制后未检测出,葡萄糖、果糖和半乳糖3种单糖含量增加;生、酒黄精低聚糖质量分数分别为78.17%、70.80%;生、酒黄精低聚糖对DPPH自由基的半数效应浓度(EC50)分别为2.12、0.41mg·mL^(-1)、ABTS+自由基的EC50分别为2.90、0.95 mg·mL^(-1);生、酒黄精低聚糖对秀丽隐杆线虫热应激和氧化应激损伤具有良好的缓解作用。结论:黄精经炮制后低聚糖成分含量发生变化;黄精低聚糖具有一定的体外和体内抗氧化作用,酒制后活性增强。以上结果可为黄精炮制前后作用改变的研究及对黄精的开发利用提供参考。

黄精霉变过程中麦角甾醇变化规律及基于电子鼻技术的快速预测模型研究

目的:麦角甾醇是霉变相关的重要指标之一,探索建立一种麦角甾醇快速检测方法。方法:采用人工模拟培养获取不同霉变程度的黄精样品;采用传统性状鉴定法开展黄精外观性状的观察、鉴定与分类;使用超高效液相色谱法进行麦角甾醇含量测定;通过电子鼻技术实现黄精气味的数字化检测。结果:共制备得到35份不同霉变程度的黄精样品;感官评价将黄精样品分为未霉变、轻微霉变和严重霉变3个阶段;35份黄精样品中麦角甾醇质量分数为0~90.4356 mg·kg^(–1),随着黄精霉变程度增加而上升;电子鼻的18根传感器中,S1~S16均表现出随着黄精霉变程度加深其最大响应值的绝对值降低,S18则相反,其最大响应值随着黄精霉变程度增加而升高。相关性分析显示,麦角甾醇含量与S1、S7~S16响应值呈极显著负相关,与S2~S6、S17、S18响应值呈极显著正相关,并且建立了回归方程Y=67.577–78.453X(r=0.634),其中Y为麦角甾醇含量、X为传感器PA/2最大响应值。结论:黄精的气味与其内在的化学成分麦角甾醇密切相关,电子鼻技术联合回归分析能为初步预测黄精中麦角甾醇的含量提供一种可行、有效的方法。