Study on Scavenging and Disconnecting Effects of Nitrosation Reaction by Extracts of Tetradium Ruticarpum and Glycyrrhizae

In order to investigate and compare the effects of Tetradium ruticarpum and Glycyrrhizae extracts in terms of scavenging sodium nitrite and inhibiting N-ni- troso compounds formation, the 1_9（3^3） orthogonal test was conducted to extract the active components. In addition, the capacities of scavenging sodium nitrite and of in- hibiting the N-nitroso compounds formation were measured to confirm the optimum condition of extracting. When the scavenging rate was considered as the main index, the maximum capacities of scavenging sodium nitrite by the extracts of Tetradi- urn ruticarpum and Glycyrrhizae were 0.836 5 and 0.558 0, respectively. Further- more, when the disconnection rate was considered as the main index, the maximum capacities of inhibiting the N-nitroso compounds formation by the extracts of Tetradi- urn ruticarpum and G/ycyrrhizae were 0.922 8 and 0.625 6, respectively. As a re- suit, the capacity of inhibiting nitrosation reaction by Tetradium ruticarpum was shown to be much stronger than G/ycyrrhizae, no matter the scavenging rate or the disconnection rate as the main index to be considered.

A systematic summary of natural compounds in Radix Glycyrrhizae

Objective： To accumulate data from studies on the compounds identified from Gancao （Radix Glycyrrhizae, GC）, andthen systematically summarize and classify these compounds according to their structural characteristics. Methods： Fivedatabases （CNKI, VIP, Wanfang data, CBM, and Pubmed） were used to search for studies on the chemical structure ofcompounds from GC. The retrieval time of the respective databases was from their inception to March 2016. Accordingto the inclusion and exclusion criteria, the papers were carefully screened, and the data were extracted. Results： A totalof 653 compounds from GC were collated from 252 articles, including flavonols （n = 201） belonging to 10 subgroups,terpenes and saponins （n = 167）, coumarins （n = 30） belonging to 3 subgroups, aliphatics （n = 206）, aromatics （n = 35）,and others （n = 14）. The flavones, represented by liquiritin and liquiritigenin, were the most reported compounds isolatedfrom GC, followed by terpenes and saponins such as glycyrrhizic acid and glycyrrhetinic acid. Conclusion： The morethan 600 natural compounds in GC may be responsible for GC’s anti-inflammatory, phlegm-expelling, cough- andpain-relieving activities.

Identification of potential anti-pneumonia pharmacological components of Glycyrrhizae Radix et Rhizoma after the treatment with Gan An He Ji oral liquid

Glycyrrhizae Radix et Rhizoma,a traditional Chinese medicine also known as Gan Cao(GC),is frequently included in clinical prescriptions for the treatment of pneumonia.However,the pharmacological components of GC for pneumonia treatment are rarely explored.Gan An He Ji oral liquid(GAHJ)has a simple composition and contains GC liquid extracts and paregoric,and has been used clinically for many years.Therefore,GAHJ was selected as a compound preparation for the study of GC in the treatment of pneumonia.We conducted an in vivo study of patients with pneumonia undergoing GAHJ treatments for three days.Using the intelligent mass spectrometry data-processing technologies to analyze the metabolism of GC in vivo,we obtained 168 related components of GC in humans,consisting of 24 prototype components and 144 metabolites,with 135 compounds screened in plasma and 82 in urine.After analysis of the metabolic transformation relationship and relative exposure,six components(liquiritin,liquiritigenin,glycyrrhizin,glycyrrhetinic acid,daidzin,and formononetin)were selected as potential effective components.The experimental results based on two animal pneumonia models and the inflammatory cell model showed that the mixture of these six components was effective in the treatment of pneumonia and lung injury and could effectively downregulate the level of inducible nitric oxide synthase(iNOS).Interestingly,glycyrrhetinic acid exhibited the strongest inhibition on iNOS and the highest exposure in vivo.The following molecular dynamic simulations indicated a strong bond between glycyrrhetinic acid and iNOS.Thus,the current study provides a pharmaceutical basis for GC and reveals the possible corresponding mechanisms in pneumonia treatment.

Intellectual Property Protection and High Quality Development of Radix Glycyrrhizae for Treatment of COVID-19

Radix Glycyrrhizae is the dominant native medicinal material variety in the north and northwestern medicinal materials producing areas.It is a main Chinese medicine of effective TCM drugs and formulas for the treatment of new coronavirus disease(COVID-19).This paper introduces the medicinal value of Radix Glycyrrhizae,involving the labeling,cultural heritage,and creative intellectual property rights of the Chinese medicinal materials,analyzes the poverty-stricken areas that are ecologically suitable for Radix Glycyrrhizae cultivation,and the superior counties and production bases of the Chinese medicinal materials.Besides,mainly from the aspects of perfect intellectual property rights,the establishment of authentic medicinal material production bases,and the construction of quality control systems,etc.,it discusses the rural revitalization strategy and the development strategy of traditional Chinese medicine(TCM),agricultural intellectual property protection and high-quality development strategy of Radix Glycyrrhizae.

Simultaneous Determination of Saponins in Radix Glycyrrhizae, Notoginseng and Ginseng by High Performance Liquid Chromatography

To establish a method for determining five saponins(notoginsenoside R1, ginsenoside Rg1, ginsenoside Re, ginsenoside Rb1 and ammonium glycyrrhizinate) in Glycyrrhizae, Notoginseng and Ginseng, the high performance liquid chromatography with diode array detector(HPLC-DAD) method was applied to an Inertsil ODS-SP column(4.6 mm×250 mm, 5 μm) with a mobile phase consisting of acetonitrile-0.05% phosphoric acid in a gradient elution manner. The flow rate was 1.0 mL/min. The column temperature was 30 ℃ and the detection wavelengths were 203 nm and 237 nm, respectively. The linear ranges were 0.700,0—7.000,0 μg for R1(r=1.000,0), 0.751,1— 7.511,4 μg for Rg1(r=1.000,0), 0.677,2—6.771,6 μg for Re(r=1.000,0), 0.733,9—7.339,1 μg for Rb1(r= 1.000,0), and 0.540,0—5.399,8 μg for ammonium glycyrrhizinate(r=0.999,9), respectively. In addition, their average recoveries were 100.28%, 105.83%, 104.09%, 99.36% and 98.54%, respectively. The relative standard deviations(RSDs) of precision, reproducibility and recovery were all less than 1.5%. The results indicate that the method is simple, accurate and reproducible so that it can be used for the simultaneous determination of the five saponins in Chinese patent medicines containing the three kinds of herbs.

Screening for cyclooxygenase 2 inhibitors from natural compounds of Radix Glycyrrhizae using computer simulation

Objective： To explore the molecular basis of the effects of Gancao （Radix Glycyrrhizae, GC） on inflammation throughthe inhibition of cyclooxygenase 2 （COX-2）. Methods： The Discovery Studio 4.5 System was used to predict thephysicochemical properties of GC molecular compounds. The Ligand Profiler was used to screen for natural GCcomponents that could combine with the COX-2 pharmacophores. The AutoDock Vina 1.1.2 software was used for themolecular docking of the natural GC components with the COX-2 protein. Results： The aromatics were the closest to thenon-steroidal anti-inflammatory drugs in terms of the three properties, namely molecular weight, molecular surface area,and molecular solubility, followed by the flavonoids; whereas the terpenoids/saponins differed most from thenon-steroidal anti-inflammatory drugs in terms of the three properties; and the aliphatics were inconsistent. One hundredand eighteen small molecules were obtained through the pharmacophore screening using GC. The molecular bindingenergy （MBE） results demonstrated that the MBE value of the flavonoids/aromatics, obtained from their binding with theCOX-2 protein, was lower than that obtained from their binding with the substrate, metabolism of arachidonic acid,whereas the MBE value of the aliphatics/terpenoids, obtained from their binding with the COX-2 protein, was higherthan that obtained from their binding with the substrate, arachidonic acid. Finally, further filtration, based on thephysicochemical properties and the molecular binding energies of the small molecules, was carried out. Forty-twonatural GC components, including 35 flavonoid and 7 aromatic constituents, with low binding energies and potentialinhibitory effects on COX-2, were screened. Conclusion： Using the three-step program, pharmacophore screening,molecular docking, and physicochemical properties analysis, we screened out 35 flavonoid molecules and 7 aromaticmolecules, which may be potential COX-2 inhibitors, from GC. Two of the 35 flavonoid molecules （licochalcone A andglabridin） have been confirmed in the laboratory to have inhibitory effects on COX-2. Our findings provide a materialbasis for the development of non-steroidal GC drugs.

甘草抗炎活性物质基础及其作用机制研究进展

炎症是机体对于刺激所产生的一种防御反应,随着炎症的发展可能导致组织的变异和增生,进一步加剧疾病的进程。甘草作为“众药之王”“国之药老”,早在《神农本草经》中就记载其清热作用较强,能“主五脏六腑寒热邪气”。现代研究表明,甘草提取物对肾小球肾炎、结肠炎症状改善明显,甘草及其制剂在治疗肝炎、胃炎、支气管炎、心肌炎、强直性脊柱炎等炎症性疾病方面亦具有一定的独特优势和发展潜力。该文旨在通过对近期国内外甘草抗炎活性物质基础及其作用机制方面的相关文献进行收集和整理,探索甘草发挥抗炎作用的主要活性成分及靶点通路,以期为甘草抗炎作用的进一步研究及抗炎药物的研发提供思路和参考。

外源钙对甘草生长及生理特性的影响

【目的】为揭示宁夏中部干旱带甘草优质高产栽培过程中甘草对不同钙浓度的适应机制,探究了外源钙对甘草生长及生理生化特性的影响。【方法】以乌拉尔甘草为试验材料,设置0(CK)、5(G_(1))、10(G_(2))、15(G_(3))、20 mmol/L(G_(4))5个钙浓度,分析比较了不同浓度外源钙处理下甘草生长、光合特性及抗氧化酶活性的变化,同时采用主成分分析和隶属函数法综合评价出适宜甘草生长的外源钙施用浓度。【结果】随着钙浓度的增加,甘草主根长、主根直径、单根鲜质量呈先增加后下降的趋势,且不同处理之间差异不显著;甘草叶片叶绿素含量、净光合速率(Pn)、蒸腾速率(Tr)、气孔导度(Gs)、胞间CO_(2)浓度(Ci)、超氧化物歧化酶(SOD)活性、过氧化氢酶(CAT)活性、过氧化物酶(POD)活性整体呈现先升后降的趋势,钙浓度为10 mmol/L时,Tr、Pn、Gs、SOD活性、CAT活性均达到峰值,且Pn、Gs显著高于对照(P<0.05)。MDA含量随着钙浓度的增加呈先下降后上升的趋势,各浓度处理间差异显著(P<0.05),钙浓度为10mmol/L时的MDA含量最低。综合评价结果表明,钙浓度为10mmol/L时,隶属函数值的均值最大,为0.739。【结论】施用外源钙有利于改善甘草的生理生化特性,促进生长发育;10mmol/L钙浓度是甘草生长的最佳施用量,可为干旱地区甘草的栽培模式提供理论依据。

甘草总黄酮对艰难梭菌的体内外抗菌作用研究

考察甘草总黄酮对艰难梭菌的抑菌活性及其抗菌机制,为艰难梭菌感染(CDI)的防治和甘草的高效利用提供依据。用微量肉汤稀释法、计数法测定了甘草总黄酮对艰难梭菌的最小抑菌浓度(MIC)、最小杀菌浓度(MBC)、生长曲线和杀菌曲线。通过扫描电镜观察、碘化丙啶(PI)摄取试验、胞外蛋白和ATP含量的测定,检测了甘草总黄酮对细菌细胞膜完整性的影响。建立了小鼠CDI模型,以体重变化和存活率为指标,评价了甘草总黄酮的体内抗菌活性。结果表明,甘草总黄酮对艰难梭菌的MIC和MBC分别为12.5μg/mL和25μg/mL,具有快速杀菌的特征。甘草总黄酮可显著破坏细菌细胞膜,导致ATP和蛋白等胞内物质的泄露,使细菌形态发生显著改变,进而导致菌体死亡。甘草总黄酮可显著缓解CDI小鼠的体重下降,7 d存活率为100%,疗效与万古霉素相当。甘草总黄酮具有显著的体内外抗艰难梭菌活性,破坏细菌细胞膜的完整性是其可能的抗菌机制。

解郁清心颗粒的定性鉴别和含量限度的制定

摸索解郁清心颗粒的定性鉴别和指标成分含量限度制定方法。采用TLC法对制剂中炒白芍、甘草进行定性鉴别;采用HPLC法对制剂中芍药苷的含量进行测定并制定含量限度。TLC用于方中药味鉴别分离度良好,供试品与对照品对应处有相同颜色的斑点,阴性无干扰。采用HPLC法测定的指标成分芍药苷的含量限度建议为≥0.8 mg/g。本实验得到的定性鉴别方法和指标成分的含量限度制定方法均具较强的专属性、准确性和可靠性,可为该制剂质控标准提供参考依据。

Effect of Glycyrrhiza Polysacchride on Growth Performance and Immunity Function of Mice

Aniso-dose glycyrrhiza polysaccharide was injected into abdominal cavity of mice. The effect of glycyrrhiza polysaccharide on growth performance and immune ruction of mice was determined. The results showed that the growth performance and immune function of mice were improved and there were significant differences among the treatment groups and control group.

α-Glucosidase Inhibitors from Glycyrrhiza uralensis Fisch.

Aim To screen for α-glucosidase inhibitor from Glyeyrrhiza uralensis Fisch.. Methods Glycyrrhizic acid, glycyrrhetinic acid, flavonoids of glycyrrhiza, alkaloids of glycyrrhiza, and glycyrrhiza polysaccharides were isolated from the root of Glycyrrhiza uralensis Fisch. respectively. Three compounds were isolated from the flavonoids of glycyrrhiza as guided by the α-glucosidase inhibitory test in vitro. Moreover, the characteristics of inhibitory kinetics of glycyrol and glycyrrhetinic acid were investi- gated. Results The flavonoids of glycyrrhiza and glycyrrhetinic acid had the strongest α-glucosidase inhibitory activity. Glycyrol,β-sitosterol and liquifitin were isolated and identified. Glycyrol was a fast- binding, reversible, noncompetitive α-glucosidase inhibitor, showing IC50 at 0.26 μg·mL^-1 Glycyrrhetinic acid was a fast-binding, irreversible α-glucosidase inhibitor, showing IC50 at 102.4 μg·mL^-1. Conclusion Glycyrol is an effective α-glucosidase inhibitor.

甘草属(Glycyrrhiza)化学成分及药理作用研究进展

文章概述了甘草目前的研究现状,扼要综述了甘草的有效成分提取方法,甘草叶及根和地上部分的化学成分。对甘草的药理作用、常见剂型及临床应用3个方面归纳总结,甘草具有补益脾气、润肺止咳、缓和药性的功效,对消化系统、心血管系统等疾病具有良好的治疗作用。并介绍了其在医疗、食品及日用化工生产中的应用。

Evidence of Hydropassive Movement in Stomatal Oscillations of Glycyrrhiza inflata under Desert Conditions

Stomatal conductance was found to change from steady-state to a slate of oscillations during daytime when vapour pressure deficit (VPD) increased to a value of 1 kPa in Glycyrrhiza inflata Batalin grown under the conditions of arid desert in north-west China. The injected metabolic inhibitors (NaN3 or carbonyl cyanide-m-chlorophenyl-hydrazone (CCCP)) slightly reduced the stomatal conductance but did not significantly decrease the intensity of stomatal oscillations (amplitude/average). The oscillation intensity was found to he significantly correlated with VPD and root resistance, but not with the respiration rate. There might exist a minimum threshold of VPD (0.8 kPa) and root resistance (1/4 relative value) that induced stomatal oscillations. These results suggested that stomatal oscillations induced by atmospheric drought stress and root resistance were mainly a type of hydropassive movement.

Effect of Different Concentrations of Mo on Yield and Accumulation of Active Constituents of Glycyrrhiza uralensis Fisch

[Objective] The aim was to explore the effects of different concentrations of molybdenum（Mo）on the growth,yield and the accumulations of active constituents of Glycyrrhiza uralensis.[Method] The seedlings of one-year-old G.uralensis were subjected to four concentrations of Mo（0,0.52,5.20 and 10.40 mg/L Mo）culturing in vermiculite and 0.52 mg/L Mo was the normal concentration in the complete Hoagland nutrition solution.The solution was irrigated to the pots every week.The fresh and dry weights of shoots and roots of the plants were measured by electronic balance.The contents of glycyrrhizic acid and liquiritin of roots were determined by HPLC.[Result] This result showed that both fresh and dry weights of the shoots and roots increased with the increasing of Mo concentrations.During the later period of this experiment,they showed significant differences among different Mo treatments.On the 105 d,both fresh and dry weights of the shoots and roots under 0 mg/L Mo reached the lowest,and the 5.20 and 10.40 mg/L Mo reached the highest.However,the contents of glycyrrhizic acid and liquiritin increased first and then decreased with increasing of Mo concentrations.During the whole experiment,the contents of glycyrrhizic acid and liquiritin under 5.20 mg/L Mo were always higher than that under other Mo treatments.[Conclusion] In conclusion,both the concentrations of 5.20 and 10.40 mg/L Mo could promote the accumulation of the shoots and roots of G.uralensis,but the 5.20 mg/L Mo was more useful for the accumulation of glycyrrhizic acid and liquiritin in the roots.

乌拉尔甘草不同组织可培养内生菌分离筛选及产β-葡萄糖苷酶菌株初筛

【目的】研究新疆阿勒泰地区乌拉尔甘草不同组织可培养内生菌的资源状况以及选育高产β-葡萄糖苷酶菌株。【方法】以乌拉尔甘草为材料,运用组织匀浆法与16S rDNA和ITS-rDNA分子生物学方法相结合方式,对其主根、须根、茎、叶中的内生菌进行分离、纯化和鉴定;采用七叶苷培养基和β-葡萄糖苷酶酶活力测定筛选产β-葡萄糖苷酶菌株。【结果】从甘草中分离获得内生菌共48株,其中内生细菌33株,内生真菌15株。在主根和茎部位分离内生细菌数量最多,内生真菌在主根中定殖的数量最多。33株甘草内生细菌隶属于8个属,芽孢杆菌属菌株分离获得最多,且甘草主根部位的内生细菌分离种类最多,在甘草不同组织中多样性表现为主根>须根=茎=叶。15株甘草内生真菌隶属于7个属,枝孢属是甘草内生真菌的优势属,内生真菌在不同组织中多样性表现为主根>茎>叶>须根。共有12株具有产β-葡萄糖苷酶的能力,其中分离自主根部位的菌株最多,占比41.7%。【结论】甘草主根组织的可培养内生菌的多样性相较其他组织丰富,且主根部位产β-葡萄糖苷酶菌株较多,是选育产酶菌株的优势部位。

硒化甘草多糖、甘草多糖及其联合抗生素对无乳链球菌体外抗菌活性及机制分析

【目的】分析硒化乌拉尔甘草多糖(selenium glycyrrhiza polysaccharide,SeGUP)、乌拉尔甘草多糖(glycymhiza polyacchiade,GUP)对无乳链球菌体外抗菌活性及机制,研究其联合的抗菌效果。【方法】通过药敏纸片筛选出对无乳链球菌敏感抗生素,采用最小二倍稀释法和棋盘法测得最小抑菌浓度(minimal inhibit concentration,MIC)、最小杀菌浓度(minimum bactericidal concentration,MBC)和联合抑菌指数(FICI index,FICI);将无乳链球菌与120、60、30 mg/mL的SeGUP、GUP及抗生素分别等量混合后绘制细菌生长速率图;通过ELISA法检测细菌培养液中碱性磷酸酶、β-半乳糖苷酶活性、蛋白质含量及胞内DNA浓度,研究SeGUP、GUP及联合抗生素对细菌细胞膜、细胞壁及DNA的影响;采用结晶紫染色法检测SeGUP、GUP及联合抗生素对无乳链球菌生物被膜形成的影响。【结果】无乳链球菌对SeGUP 480 mg/mL、GUP 480 mg/mL均呈现中度敏感,抑菌圈直径为14.0、11.0 mm;无乳链球菌对盐酸卡那霉素、盐酸多西环素、头孢曲松钠、氟苯尼考极敏;SeGUP和GUP分别对肺炎链球菌的MIC、MBC为120、240 mg/mL和240、480 mg/mL;SeGUP与盐酸卡那霉素、盐酸多西环素、头孢曲松钠、氟苯尼考的FICI为1.5、0.75、1、0.5,SeGUP与盐酸卡那霉素之间关系为无关,与盐酸多西环素、头孢曲松钠之间为相加关系,与氟苯尼考间为协同关系,GUP与盐酸卡那霉素、盐酸多西环素、头孢曲松钠、氟苯尼考的FICI为1.5、1、1.5、0.75,GUP与盐酸卡那霉素、头孢曲松钠之间关系为无关,与盐酸多西环素、氟苯尼考间为相加关系;SeGUP、GUP明显抑制细菌的生长,其中SeGUP 120 mg/mL效果最强,SeGUP、GUP与盐酸多西环素、头孢曲松钠、氟苯尼考联合使用后增强了抑制效;SeGUP、GUP及联合抗生素使用后均能显著升高培养液中碱性磷酸酶(AKP)、β-半乳糖苷酶(β-gal)、蛋白质含量(P<0.05),显著降低胞内DNA含量(P<0.05),显著抑制无乳链球菌生物被膜的形成(P<0.05)。【结论】SeGUP能够通过损伤细菌细胞壁、细胞膜完整性,影响DNA含量对无乳链球菌产生明显的抑制作用,且其抗菌活性优于GUP。SeGUP与抗生素联合使用后能够增强抗生素的抗菌活性,并且其作用效果强于单独使用SeGUP、GUP和抗生素。

甘草属Glycyrrhiza L.植物资源化学及利用研究进展

甘草作为一种优良的天然植物资源,已被广泛应用于医药、食品、化工、畜牧等领域,国内外相关研究也方兴正艾,但近年来随着生态环境的破坏及无节制的采挖,资源状况日益严峻。本文从中药资源化学的角度出发,对甘草属植物的种类及资源分布进行总结,并针对不同品种甘草的活性成分、药理作用以及应用现状进行综述。旨在为进一步深入研究和开发该类药用植物资源提供参考,并为多层次、精细化的甘草资源综合利用提供依据。

多基原甘草质量标准探讨

目的分析多基原甘草质量现状及存在的问题,探索制约因素并提出解决方案。方法通过和企业座谈、实地调研、文献查询及甘草质量标准研究工作,分析当前甘草产业现状、药用标准问题及发展契机。结果以《中华人民共和国药典》(2020年版)含量测定标准,我国胀果甘草、光果甘草中甘草苷含量不合格率偏高,但3种基原的甘草存在其他具有药理作用的黄酮类化学成分,仅以甘草苷作为质量控制指标评价不够全面。结论建议进一步提高和完善《中华人民共和国药典》中胀果甘草和光果甘草质量标准,采用多种黄酮类成分作为甘草质量评价指标,进而推进我国甘草种植业发展。

SCoT与EST-SSR标记检测甘草属(Glycyrrhiza L.)植物遗传多样性的比较研究

为探讨SCoT、EST-SSR标记技术在甘草属植物遗传多样性研究的适用性,采用SCoT、EST-SSR分子标记对甘草属8自然居群的4种1变种共计14个个体进行基因组DNA的多态性检测。实验筛选出的18对SCoT通用引物共扩增出313条带,其中312条为多态性条带,平均多态率(PPB)为99.68%,平均有效等位基因数(Ne)和位点品均期望杂合度(He)等遗传参数与EST-SSR标记无显著差异。多态性检测效率高的标记为SCoT,E=17.333,Ai=21.145,高于EST-SSR,E=6.909,Ai=8.719。SCoT与EST-SSR标记聚类分析结果趋势相似,均将14份材料划分为3组,与基于形态学分类结果一致。2种标记相比,EST-SSR(0.820)产生的平均相似系数范围大于SCoT(0.785),EST-SSR标记个体间遗传差异检测能力略高于SCoT。结果表明SCoT和EST-SSR两种标记均可揭示甘草属种间与种内的遗传多样性水平以及亲缘关系,是进行甘草属植物自然群体的遗传结构、种间基因渐渗等研究的有效分子标记。其中,SCoT标记多态性高,信息量大,更适于甘草属植物种质资源的遗传多样性分析;个体间遗传差异检测上EST-SSR标记效果更佳,更利于疑难种鉴定及亲本分析。