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.

Measurement and comparison of glycyrrhizic acid contents in root of licorice(Glycyrrhiza uralensis Fisch.)from different cultivating areas

The samples of licorice (Glycyrrhiza uralensis Fisch.) from 14 different cultivated areas were determined by the method of high Performance Capillary Electrophoresis (HPCE) for the contents of glycyrrihizic acid (GA) in root. The results showed that the licorice plants come from various cultivated areas of China has different contents of GA. The GA content of licorice from Zhaodong in Heilongjiang Province is the highest, followed by those from E抰uoke, Chifeng, and Hangjin Banner in Inner Mongolia. Some suggestions for establishing the production base of licorice were put forward based on the study.

Primary Speciation Analysis on 6 Kinds of Microelements in Glycyrrhiza uralensis Fisch.

[Objective]The aim was to analyze the primary speciation of 6 microelements in Glycyrrhiza uralensis Fisch. and provide theoretical basis for explaining pharmacodynamic principle of liquorice and discussing quality control of liquorice planting. [Method]The 6 elements Cu,Zn,Ca,Fe,Mg and Mn in roots of G.uralensis were extracted based on traditional decoction method and were separated into water-soluble state and suspension state by micro porous filtering film. The elements in water-soluble state were detected by flame atomic adsorption spectrophotometry （FAAS）. [Result]The results showed that extractive rates of the elements were in the range of 1.71%-60.06%,and immerse-residue ratio in 0.018 3-1.682 0; the results also indicated that the immerse-residue ratio of Zn was biggest （1.68）,Zn played an important medical role and might be considered as the best characteristic element in G.uralensis; the recoveries of the elements were ranged from 95.72% to 103.15% and relative standard deviations （RSD） were less than 2.38%. [Conclusion]Because of its high accuracy,FAAS method is feasible for analyzing primary speciation of microelements in G.uralensis.

Effect of Glycyrrhiza uralensis Fisch polysaccharide on growth performance and immunologic function in mice in Ural City,Xinjiang

Objective:To discuss the effect of Glycyrrhiza uralensis(G.uralensis) Fisch polysaccharide on growth performance and immunologic function in mice in Ural City,Xinjiang and to provide important data supporting the application of Glycyrrhiza polysaccharide.Methods:A total of100 Kunming mice aged 3 weeks old were randomly divided into 5 groups with 20 mice in each group(10 were females and 10 were males).About 0.5 mL normal saline was given to the mice of control group every day and 0.5 mL G.uralensis Fisch polysaccharide was given to the mice of other groups at the concentration of 1,20,50 and 100 mg/mL respectively.The growth performance(average body weight,average daily feed intake and feed efficiency),immune organ indexes(spleen index and thymus index) and immunologic function(serum IL-2,CD4^+/CD8^+ and the activity of NK cells) of mice in each group were detected continuously.Results:The average body weight,feed efficiency,serum IL-2,CD4^+/CD8^+ and the activity of NK cells of mice were increased with the increase of administrated time after administrating G.uralensis Fisch polysaccharide and were reached up the largest level on Day 28.At the same time,each index was proportional to the given dose and was significantly higher than those of control group and reached up the largest level at the administrated dose of 100 mg/mL.After administrating G.uralensis Fisch polysaccharide,the spleen index and thymus index of mice were increased with the increase of administrated dose and the spleen index and thymus index of mice administrated with the dose of 100 mg/mL were maximum which was more than 1.51 times and 1.43 times of that in control group respectively and the comparative differences showed statistical significance(P<0.05).The average daily feed intake of mice in each group was increased with the passage of lime and at the same time,the comparison of average daily feed intake of mice in each group was not significantly different(P>0.05).Conclusions:G.uralensis Fisch polysaccharide can significantly improve the growth performance and immunologic function of mice and laid a research basis for the clinical application of G.uralensis Fisch polysaccharide.

乌拉尔甘草(Glycyrrhiza uralensis Fisch.)大面积人工种植试验初报

人工种植甘草是治沙工程中的内容之一 ,试验通过采取机械直播的方式种植甘草 ,研究了不同播种深度、灌溉水平、种植密度对出苗率、保苗率及生长量的影响。结果表明 ,在灌溉条件下 ,甘草播种量为 2 2 .5kg/hm2 ,密度 30万株 /hm2 ;在喷水条件下 ,喷水时数达 2 0h,灌溉总额约为 1 50 0 m3/hm2

Molecular Cloning and Expression Analysis of an Actin Gene from Chinese Licorice(Glycyrrhiza uralensis Fisch)

A PCR-based homologous cloning strategy was used to identify an actin gene from the roots of Chinese licorice（Glycyrrhiza uralensis Fisch）. Results of sequence analysis indicate that a 1137 bp cDNA with an open reading frame encoding 377 amino acids, actin ortholog, GuActin, was successfully cloned and characterized（GenBank accession No. EU190972）. Thus far, GuActin is the first actin of Chinese licorice that has been identified at a molecular level. Analysis by Northern blot shows that GuActin was expressed strongly in the roots, particularly in radicles than in stems and leaves. These results suggest that GuActin may be a member of the vegetative subfamily of the actin family.

Effect of Ar Ion Beam Implantation on Morphological and Physiological Characteristics of Liquorice(Glycyrrhiza uralensis Fisch)Under Short-Term Artificial Drought Conditions

Ar^＋ ion beam with low energy of 30 keV was implanted into liquorice （Glycyrrhiza uralensis Fisch） seeds at the doses of 0, 600, 900 and 1200 × （2.6 × 10^13） ions/cm^2, respectively. The seeds were sowed in pots and after one month the plants were subjected to different drought conditions for two months. Then the plants＇ morphological and physiological characteristics, antioxidation enzymes and levels of endogenous hormones were investigated. The results showed that ion implantation at a proper dose can greatly enhance the liquorice seedlings＇ resistance against drought stress.

Impact of Ion Implantation on Licorice(Glycyrrhiza uralensis Fisch) Growth and Antioxidant Activity Under Drought Stress

Low energy ion beams are known to have stimulation effects on plant generation and to improve plants＇ intrinsic quality. In the present study, the growth and physiological index of licorice implanted with 0, 8, 10; 12 and 14× （2.6× 10^15） ions/cm^2 were investigated under well-watered and drought-stress conditions. The results showed that a proper dose of ion implan- tation was particularly efficient in stimulating the licorice growth and improving the plant biomass significantly in both the well-watered and drought-stress conditions. The physiological results of licorice measured by leaf water potential, lipid oxidation, soluble protein and antioxidant system showed a significant correlation between ion implantation and water regime except for leaf water potential. Therefore, the study indicated that ion implantation can enhance licorice＇s drought tolerance by increasing the activity of superoxide dismutase （SOD）, eatalase （CAT） and DPPH （1,1-diphenyl-2-picrylhydrazyl） radical scavenging ability to lower oxidative damage to lipids in plants. Ion beam implantation, therefore, provides an alternative method to enhance licorice drought tolerance.

Effects of Different Arbuscular Mycorrhizal Fungi on Growth and Protective Enzyme Activity of Glycyrrhiza uralensis

[Objective] This study aimed to investigate the effects of 5 kinds of arbuscular mycorrhizal fungi( Acaulospora mellea,Glomus mosseae,Glomus versiforme,Glomus aggregatum,Glomus etunicatum) on the growth and protective enzyme activity of Glycyrrhiza uralensis Fisch. [Method] The growth indicators and protective enzymes activity of glycyrrhiza plants inoculated and uninoculated with fungi were compared. [Result] The plant height,basal diameter,main root length,aboveground fresh weight,underground fresh weight,aboveground dry weight and underground dry weight of the inoculated glycyrrhiza plants were increased significantly compared with those in the control( non-inoculation) group. In the inoculated glycyrrhiza plants,the growth index was significantly increased compared with that in the control group( P <0. 05); the activities of superoxide dismutase( SOD) and peroxidase( POD) increased first and then decreased; and the activity of catalase( CAT) showed a continuous rising trend. The effects of different inoculants on the growth of G. uralensis were significantly different.[Conclusion]G. etunicatum,G. mosseae and G. aggregatum had a significant effect on the growth of G. uralensis,and were superior to other fungi in resisting the adverse environment.

INHIBITORY EFFECT OF GLYCYRRHIZA URALENSIS FISH AND CHELIDONIUM MAJUS L ON GASTROCARCINOGENESIS . INDUCED BY N-METHYL-N' -NITRO-N-NITROSOGUANIDINE

In order to investigate the antagonistic effect of Glycyrrhlza Uralensis Fish (GUF) and Chelidonium maJus L (CML) on gastrccarcinogenesls Induced by MNNG in Wastar rats, we treated the rats with MNNG alone (group 1) and with MNNG plus GUF and CML (group 2 and 3) respectively. The Incidence of Infiltrating adenocarcinoma of the glandular stomach and duodenum in group 2 was significantly lower than that in group 1 (26.7% vs. 67.8%). The differentiation and aggresslvenees of carcinomas occured in group 2 were much better and mild than those in group 1. Present study also demonstrated that the Inhibitory effect of CML on proliferation of human stomach carcinoma cell line MGC-803 was very remarkable; in addition, GUF and CML were able to antagonise the mutagenlc activation of MNNG. These results suggest that GUF and CML may be empoyed In prevention of gastric carcinoma.

双波长HPLC法同时测定消炎灵胶囊中四种成分的含量

目的:建立双波长HPLC法同时测定消炎灵胶囊中苦玄参苷Ⅰ_(A)、苦玄参苷Ⅰ_(B)、甘草苷、甘草酸含量,对市售消炎灵胶囊中苦玄参和甘草质量进行研究与评价。方法:使用HPLC,选择Waters Xselect C_(18)色谱柱,以乙腈为流动相A,0.1%磷酸溶液为流动相B进行梯度洗脱,流速1.0 mL/min,柱温30℃,检测波长237 nm(甘草苷、甘草酸)、264 nm(苦玄参苷Ⅰ_(A)、苦玄参苷Ⅰ_(B))。结果:65批消炎灵胶囊4种成分线性关系良好(r=0.9998~1.0000),平均加样回收率为97.6%~98.6%(RSD值均小于1%)。部分企业消炎灵胶囊不同批次间4种成分含量存在较大差异,说明各批次间均一性较差。提示各生产企业加强对药材质量控制。结论:该方法简单快捷,可同时测定4个成分含量,并具有良好精密度、重复性和稳定性,为消炎灵胶囊中苦玄参苷Ⅰ_(A)、苦玄参苷Ⅰ_(B)、甘草苷、甘草酸质量评价提供参考。

HPLC-DAD同时分析甘草中7种有效成分

目的采用HPLC—DAD中梯度洗脱和检测波长时间序列采样的方法，建立甘草（Glycyrrhiza uralensis Fisch．）中4种黄酮成分（甘草苷、异甘草苷、甘草素、异甘草素）和3种甘草皂苷成分（甘草酸、甘草皂苷G2、乌拉尔甘草皂苷B）的同时分析方法。方法采用C18（4．6mm×250mm，5um）色谱柱。流动相：乙腈-（0．5％醋酸铵＋1％冰醋酸），梯度洗脱0～12min从6：94到22：78，12～25min从22：78到25：75，25—45min从25：75到40：60，45～50min从40：60到6：94，保持5min；检测波长：时间序列采样。结果7种成分的线性关系良好，平均加样回收率在98％～103％之间。结论该方法准确、灵敏度高，可用于甘草药材质量的评价。

甘草野生种群遗传多样性的AFLP分析

【目的】甘草具有重要的药用、工业和生态价值,目前处于濒危状态,进行遗传多样性研究可以为甘草资源的保护和利用奠定基础。【方法】利用AFLP分子标记对来自中国甘草主产区的16个野生种群共320个单株进行遗传多样性研究。【结果】(1)利用15对AFLP引物共扩增出759条谱带,其中多态性谱带527条,多态性条带百分率为69.43%;(2)Nei’基因多样性指数为0.13～0.19,种群总体多样性指数为0.25;Shannon多态性信息指数的变异范围在0.19～0.28,总体为0.39;宁夏地区甘草种群遗传多样性水平最高,甘肃酒泉种群的遗传多样性水平最低。(3)AMOVA分析表明甘草种群间的遗传变异占总变异的18.64%,种群内变异占67.16%。利用UPGMA聚类可将供试16个群体划分为3类,聚类结果表现出明显的地域性。【结论】该研究明确了中国野生甘草遗传多样性处于中等偏下水平,种群内广泛的变异能够为野生资源保护和良种选育提供理论依据。

外源甘草酸对NaCl胁迫条件下甘草幼苗生长、根部甘草酸含量以及几种与盐胁迫相关生理指标的影响

50和100mmol·L-1NaCl有利于甘草幼苗根部甘草酸积累。在200mmol·L-1NaCl胁迫下喷施甘草酸(5mmol·L-1),可以明显促进根的生长,提高叶片中叶绿素含量和叶绿素荧光参数Fv/Fm,降低叶片电导率、可溶性糖和脯氨酸含量,增加根部甘草酸含量。

HPLC法分析12产地甘草中4种主要黄酮类化合物的含量

目的:分析不同产地甘草中甘草苷、异甘草苷、甘草素、异甘草素的含量,为不同产地甘草的质量评价提供参考。方法:从全国7省区12产地采集甘草种子,栽培于北京中医药大学药草园,1年后每产地随机挑取15株甘草作为实验材料,利用ITS序列鉴定其基原,采用HPLC法测定其甘草苷、异甘草苷、甘草素和异甘草素的含量,分析各成分含量的差异性及相关性。结果:ITS鉴定结果表明甘草样品均为乌拉尔甘草(Glycyrrhiza uralensis Fisch.)。HPLC分析结果显示:甘草苷的标准曲线为Y=0.000 5X+1×10^(-5)(R^2=0.999 9),在0.113 7μg^0.854 1μg范围内线性良好;异甘草苷的标准曲线为Y=0.000 3X-3×10^(-5)(R^2=0.999 9),在0.025 3μg^0.204 8μg范围内线性良好;甘草素的标准曲线为Y=0.000 3X+1×10^(-4)(R^2=0.999 8),在0.009 3μg^0.092 7μg范围内线性良好;异甘草素的标准曲线为Y=0.000 2X-7×10^(-5)(R^2=0.999 9),在0.004 6μg^0.030 7μg范围内线性良好。不同产地甘草中4种黄酮类成分含量差异显著,其中山西应县甘草样品的甘草苷、异甘草苷、甘草素及异甘草素的含量均为最高,而黑龙江肇州县、新疆尼勒克县、额敏县、阿拉尔县甘草样品的甘草苷、异甘草苷、甘草素及异甘草素的含量普遍较低。且所有样品中4种黄酮类化合物的含量之间均存在显著相关关系。结论:不同产地甘草质量差异性显著,本实验结果可以为不同产地甘草的质量评价提供参考。

甘草HPLC指纹图谱研究

目的:建立乌拉尔甘草(Glycyrrhiza uralensis Fisch)的HPLC指纹图谱。方法:采用反相高效液相色谱法,选用APOLLOC_8柱(250 mm×4.6 mm,5μm),甲醇-0.5%冰醋酸为流动相梯度洗脱,采集时间为80 min,流速为1.0 ml·min^(-1)。结果:建立甘草药材的HPLC指纹图谱,标定甘草药材14个共有峰,方法学考察结果符合指纹图谱技术要求。结论:该方法稳定,准确,可靠,为今后甘草药材质量控制提供科学依据。

内蒙古产甘草药材HPLC指纹图谱研究

目的研究乌拉尔甘草(Glycyrrhiza uralensis Fisch.)药材的指纹图谱测定方法,建立特定产地甘草药材的HPLC标准指纹图谱。方法采用反相高效液相色谱法,选用PhenomenexC18柱(4.6mm×250mm,5μm),乙腈-醋酸盐缓冲溶液为流动相梯度洗脱,运用梯度波长采集法,采集时间为60min,流速为1mL/min。结果建立甘草药材的HPLC指纹图谱,标定了甘草药材10个共有指纹峰,方法学考察结果符合指纹图谱技术要求。结论该方法稳定、准确、可靠,为甘草药材质控标准的制定提供了科学依据。

^(60)Co-γ辐射对NaCl胁迫环境下乌拉尔甘草种子萌发的影响

用不同剂量(0~300 Gy)的60Co-γ射线辐照乌拉尔甘草种子,研究辐照处理对乌拉尔甘草种子在不同浓度NaCl(0~400 mmol/L)胁迫环境下萌发率和累积发芽率的影响,以期筛选出有助于提高乌拉尔甘草种子耐盐性能的辐射剂量。结果表明:随着NaCl浓度的增加,种子萌发率与NaCl浓度之间呈负相关。乌拉尔甘草种子萌发的"滞后"效应有加剧的趋势,主要表现为发芽率的降低、起始萌发时间的推迟以及达到最大发芽率所需时间的延长。但经100 Gy和300Gy的60Co-γ射线辐照处理的乌拉尔甘草种子,不仅提高了其在NaCl胁迫下的萌发率,而且缩短了种子的初始萌发时间,对NaCl胁迫表现出了较好的耐受性。

HPLC-ELSD法测定甘草中单糖的含量

建立一种高效快速的高效液相色谱-蒸发光散射检测法(HPLC-ELSD)测定中药甘草中单糖———果糖及蔗糖含量的方法。采用CHROMATOREXNH2柱为固定相;以乙腈∶水为85∶15(V/V)为流动相;流速1.0mL/min;漂移管温度为84℃;氮气流速为2.5L/min。在上述色谱条件下,果糖和蔗糖的线性范围分别为1.422～3.318μg及12.028～60.140μg;且该二种单糖的检测限分别(信/噪=3)为4.74ng和3.31ng。该方法用于传统中药甘草中单糖的分离及含量测定,结果令人满意。

黄药子配伍甘草对LO2细胞膜通透性影响的研究

目的:研究甘草配伍能减轻黄药子对人肝细胞(LO2)膜通透性的影响。方法:激光扫描共聚焦显微镜(LSCM)对经二乙酸荧光素(FDA)染色后的LO2细胞进行实时动态荧光强度检测。通过比较荧光强度的降低率来考察黄药子及其甘草配伍后含药血清在短时间内对细胞膜通透性的影响。结果:LSCM记录细胞荧光强度变化,在10 min扫描停止时,黄药子各组荧光强度降低率与空白组有统计学差异(P<0.05),而甘草配伍组对荧光强度的影响较黄药子组有统计学差异(P<0.05),与空白组无差异(P>0.05)。结论:黄药子含药血清短时间内即影响肝细胞膜通透性,甘草配伍后可明显减弱损伤作用,黄药子引起肝损伤的机制可能是其对肝细胞膜的直接损伤作用,甘草配伍减毒作用与此途径有关。