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.

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.

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.

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

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.

Ethanol Extract of Glycyrrhiza uralensis Fisch:Antidiarrheal Activity in Mice and Contraction Effect in Isolated Rabbit Jejunum

Objective:To evaluate the antidiarrheal effect of ethanol extract of Glycyrrhiza uralensis Fisch root(GFR)in vivo and jejunal contraction in vitro.Methods:In vivo,50 mice were divided into negative control,positive control(verapamil),low-,medium-and high-dose GFR(250,500,1,000 mg/kg)groups by a random number table,10 mice in each group.The antidiarrheal activity was evaluated in castor oil-induced diarrhea mice model by evacuation index(El).In vitro,the effects of GFR(0.01,0.03,0.1,0.3,1,3,and 10 g/L)on the spontaneous contraction of isolated smooth muscle of rabbit jejunum and contraction of pretreated by Acetylcholine(ACh,10μmol/L)and KCl(60 mmol/L)were observed for 200 s.In addition,CaCl_(2)was accumulated to further study its mechanism after pretreating jejunal smooth muscle with GFR(1 and 3 g/L)or verapamil(0.03 and 0.1μmol/L)in a Ca^(2+)-free-high-K+solution containing ethylene diamine tetraacetic acid(EDTA).Results:GFR(500 and 1,000 mg/kg)significantly reduced El in castor oil-induced diarrhea model mice(P<0.01).Meanwhile,GFR(0.01,0.03,0.1,0.3,1,3,and 10 g/L)inhibited the spontaneous contraction of rabbit jejunum(P<0.05 or P<0.01).Contraction of jejunums samples pretreated by ACh and KCI with 50% effective concentration(EC50)values was 1.05(0.71-1.24),0.34(0.29-0.41)and 0.15(0.11-0.20)g/L,respectively.In addition,GFR moved the concentration-effect curve of CaCl_(2)down to the right,showing a similar effect to verapamil.Conclusions:GFR can effectively reduce diarrhea and inhibit intestinal contraction,and these antidiarrheal effects may be based on blocking L-type Ca^(2+) channels and muscarinic receptors.

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。

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。

大枣生姜甘草复合饮料制备及抗氧化研究

以大枣、甘草、生姜为原料,研制一种复合饮料.利用单因素分析和正交试验,以感官评分和可溶性固形物含量为评价指标,研制枣姜甘草复合饮料的最优配方.研究表明,枣姜甘草复合饮料的最佳配方为:大枣汁54%、生姜汁14%、甘草汁5%,柠檬酸0.025 g.所得复合饮料感官评分为82.67,总糖含量为1.744 g/100g,总酸度为0.533 g/kg,可溶性固形物含量为11.50%, DPPH自由基清除率为77.08%,羟自由基清除率为24.20%,制备得到的产品具有良好的抗氧化能力.

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。

基于化学计量学建模分析对甘草质量的综合评价

为探讨主成分分析法和相关性分析综合评价不同种植地点和时间甘草药材质量的可行性,分别对甘肃省张掖市不同地区种植的共14批甘草药材中芹糖甘草苷、甘草苷、甘草酸、浸出物、多糖、总黄酮、粗纤维及水分含量进行测定,并对测定的结果进行相关性分析及主成分分析。相关性分析表明,各指标存在潜在的相关性,其中甘草浸出物与总黄酮、甘草酸之间呈极显著性相关,甘草苷与甘草酸之间呈显著性相关,总黄酮与芹糖甘草苷、甘草苷之间呈显著性相关,芹糖甘草苷与甘草苷之间呈显著性相关,其他指标之间的相关性显著性弱或不显著。综合主成分评价表明,14号样品品质最优,肃南县种植的甘草质量普遍较高。综上表明相关性分析及主成分分析可以用于不同地点甘草的质量分析及综合评价,可以为甘草的多指标评价提供借鉴,也可以为进一步建立全面的甘草质量评价体系奠定基础。

Synthesis of amino-silane modified magnetic silica adsorbents and application for adsorption of flavonoids from Glycyrrhiza uralensis Fisch

Magnetic separation technology was applied in the separation of flavonoids from the licorice root in this work. Licorice flavonoids (LF) displayed a remarkable array of biological and pharmacological activities. The magnetic adsorbents with functional -NH2 groups were synthesized by immobilization of amino-silane on the surface of the magnetic silica supports, which were prepared by co-precipitation method. The adsorption and desorption characteristics of the magnetic adsorbents for the separation of LF have been evaluated. The purity of an enriched extract with this method was 16.7% while the crude extract only had about 6.8% purity. Therefore, it can be concluded that these kinds of magnetic adsorbents have selectivity to the flavonoids to some extent. The affinity selectivity of the adsorbents is based on the formation of hydrogen bonding between the -NH2 on the magnetic adsorbents and -OH,-CO on the flavonoids.

甘肃省干旱山区甘草产业现状及发展对策

通过调研甘肃省榆中县北部干旱山区甘草产业生产现状,在分析甘草生产中种苗成本高、本土化育苗滞后、机械化水平低、劳动力匮乏、甘草加工工艺落后等问题基础上,提出了甘肃干旱山区甘草产业发展应加强新品种引进、开展本土化育苗和栽培技术研究、提高机械化程度、发挥龙头企业引领作用、提升深加工能力、培育产品品牌效应等建议,为甘肃省干旱山区甘草产业健康发展提供理论指导。

基于UPLC-LTQ-Orbitrap MS探究生、炙甘草黄酮类成分及其抗氧化活性差异

目的:探究甘草蜜炙前后黄酮类成分变化与其抗氧化活性的关系。方法:提取蜜炙前后甘草总黄酮,采用紫外分光光度法、超高效液相色谱串联线性离子阱高分辨质谱法分别进行含量测定及成分分析,采用总抗氧化能力测定法、超氧阴离子自由基清除法、羟自由基清除法及1,1-二苯基-2-三硝基苯肼(DPPH)自由基清除法4种体外方法测定其抗氧化活性。结果:甘草蜜炙后总黄酮抗氧化活性明显提高,总黄酮含量较生品降低;质谱结果显示,生、炙甘草共有成分中甘草查耳酮A等10个黄酮类成分含量上升,反曲刺酮素B等5个成分含量下降,其中甘草查耳酮A含量提升最显著。结论:蜜炙甘草总黄酮抗氧化活性的提高可能主要在于生、炙甘草黄酮成分含量的差异,其中甘草查耳酮A含量的变化对总黄酮抗氧化活性的提升起重要作用,为甘草“生熟异用”研究提供了依据。

不同光质对甘草苗期生长及活性成分含量的影响

目的:探究不同光质对甘草苗期生长及活性成分积累的影响特征。方法:采用白光(对照)、红光、蓝光、红蓝光(红光∶蓝光=2∶1)和蓝红光(蓝光∶红光=2∶1)5种光质处理甘草,分别在光质处理36、106 d测定甘草的生长指标,建立甘草根部12个活性成分含量的测定方法,结合聚类分析探析不同光质对甘草活性成分积累的影响特征。结果:红光促进甘草株高增长的作用较其他光质大,红光下甘草株高比对照高40.9%~45.1%;红蓝光促进甘草根部生长的作用明显,不同光质下甘草根长、根粗和根干质量增长量变化趋势一致,均为红蓝光>红光>白光>蓝红光>蓝光;不同光质处理106 d甘草的根冠比均大于处理36 d,红蓝光处理106 d甘草的根冠比最大,是对照的2.8倍。聚类分析结果表明,甘草的活性成分随不同光质处理时间的延长而增大,不同光质下甘草酸、芹糖异甘草苷、异甘草苷等9个活性成分含量随不同光质处理时间的增加呈升高趋势。红蓝光促进甘草中甘草酸、芹糖异甘草苷、异甘草苷、异甘草素、新异甘草苷、甘草素和甘草苷积累的作用较其他光质大,分别比对照高88.24%、17.39%、284.31%、156.95%、246.35%、89.06%和207.40%;蓝红光促进甘草中甘草苷、甘草素、甘草酸和异甘草素积累的作用较大,分别比对照高12.42%、3.00%、7.07%和5.41%;增加蓝光比例有利于甘草中甘草查耳酮A的积累,红光处理36 d促进山柰酚和刺甘草查耳酮的积累,处理106 d抑制以上2个成分的积累。结论:红光对甘草地上部生长有较强促进作用,红蓝光可以显著促进甘草根系的生长和生物量积累;红光和蓝光混合光质较红光或蓝光单色光质对甘草中多种活性成分积累的促进作用更大,以红蓝光处理更佳,红蓝光与甘草中异甘草素、甘草素和甘草苷的积累呈正相关。为光质在甘草次生代谢成分的定向培育应用提供参考。

γ-氨基丁酸对甘草种子萌发及呼吸代谢的影响

以甘草种子为材料,测定不同浓度的γ-氨基丁酸(GABA)处理及GABA抑制剂处理后甘草种子的动态质量、吸胀速率、发芽率、发芽指数、呼吸速率、活力指数以及呼吸代谢物有机酸和氨基酸的含量,探究GABA对甘草种子萌发及呼吸代谢的影响。结果表明,外源GABA处理在一定程度上能够提高甘草种子的萌发指数、活力指数、质量和呼吸速率,其中,2 mmol/L GABA处理的种子活力指数和萌发指数较高,分别是对照组的1.45倍和1.38倍(p<0.05),处理48 h时质量为对照的1.05倍;一定浓度外源GABA处理可以提高种子内部GABA、琥珀酸等呼吸相关代谢物的含量;一定浓度GABA抑制剂处理能够降低甘草种子的呼吸速率和增加种子质量,48 h时的呼吸速率为对照组的92.86%,抑制剂同时可以降低延胡索酸和琥珀酸的含量。研究表明,一定浓度的GABA处理有利于甘草种子萌发,能够提高种子内源GABA以及其他呼吸代谢物的含量,促进三羧酸循环。