Phylogenetic relationship of six Glycyrrhiza species based on rbcL sequences and chemical constituents

The nucleotide sequences of ribulose-1,5-bisphosphate carboxylase/oxygenase large subunit gene (rbcL) of Glycyrrhiza glabra, G. uralensis, G. inflata, G. echinata, G. macedonica and G. pallidiflora have been determined to construct their phylogenetic tree. Based on these sequences, the six Glycyrrhiza species were divided into two groups: three, G. glabra, G. uralensis, and G. inflata, which produce glycyrrhizin as a major saponin, and the others, G. echinata, G. macedonica and G. pallidiflora, which produce macedonoside C as a major saponin. Among the three glycyrrhizin-producing species, only two nucleotide substitutions were observed between the rbcL sequences of G. glabra and G. uralensis, and the sequence of G. uralensis was identical to that of G. inflata, indicating that G. uralensis and G. inflata are closely related. Among the three macedonoside C-producing species, only one nucleotide substitution was observed between those of G. echinata and G. macedonica, indicating that these two species are also closely related.     

光果甘草三萜皂苷类化学成分研究

采用聚酰胺—大孔树脂柱色谱、ODS中压柱色谱以及半制备液相色谱分离技术,对光果甘草(Glycyrrhiza glabra L.)水提取物中三萜皂苷类成分进行分离纯化,共获得10个三萜皂苷类化合物,依据理化性质及NMR、MS波谱数据鉴定化合物结构,分别鉴定为3β-O-[β-D-glucuronpyranosyl-(1→2)-β-D-glucuronpyranosyl]-30β-O-β-Dglucuronpyranosyl-oleanane-11-oxo-12(13)-ene (1)、3β-O-[β-D-glucuronpyranosyl-(1→2)-β-D-glucuronpyranosyl]-30β-O-α-L-rhamnopyranosyl-oleanane-11-oxo-12(13)-en-22β,30-diol (2)、uralsaponin C (3)、licorice-saponin A3 (4)、licoricesaponin P2 (5)、22β-acetoxyl-glycyrrhizin (6)、macedonoside A (7)、29-hydroxyl-glycyr...     

Bioactive constituents of oleanane-type triterpene saponins from the roots of Glycyrrhiza glabra

Three new oleanane-type triterpene saponins, namely licorice-saponin M3 (1), licorice-saponin N4 (2), and licorice-saponin O4 (3), an artificial product (4), as well as five known triterpene glucuronides (5–9), were isolated from the roots of Glycyrrhiza glabra L. Their structures were established using 1D and 2D NMR spectroscopy, mass spectrometry, and by comparison with spectroscopic data reported in the literature. The inhibitory effects of the selected compounds on neuraminidase were evaluated, and the preliminary structure–activity relationship was also predicted.     

甘草中三种皂甙类成分的高效液相色谱法分离和含量测定

本文报道应用高效液相色谱法对甘草中三种皂甙类成分甘草酸(glycyrrhizic acid,S-Ⅰ),乌拉甘草皂甙甲(uralsaponin A,S-Ⅲ)和乌拉甘草甙乙(uralsaponin B,S-Ⅱ)进行分离和含量测定。采用ODS柱,乙腈—3%乙酸(47:53)为流动相,248 nm为检测波长,三种皂甙分离良好,可在20 min内完成测定。     

甘草及其活性成分的表面活性比较研究

目的 研究甘草及其活性成分的表面活性,为更合理地研究甘草中成分的增溶能力提供参考.方法 在不同温度下,采用Wilhelmy吊片法测定溶液的表面张力和溶液的临界胶束浓度(CMC).结果 分析了溶液的表面活性效能、效率和表面过剩浓度,甘草水煎液表面活性较好.结论 甘草中除皂苷组分具有表面活性外,其他组分对表面活性有贡献.     

不同产地甘草有效成分含量分析

目的对不同产地甘草中甘草酸、甘草苷以及甘草总皂苷进行含量测定。方法采用高效液相色谱法测定甘草酸、甘草苷的含量。色谱柱为Thermo sentifi c ODS-2 Hypersil,流动相为乙腈-0.05%磷酸;以甘草酸单铵盐为对照品,香草醛-高氯酸为显色剂,比色法测定甘草总皂苷含量。结果测得不同产地不同种植方式甘草中甘草酸和甘草苷含量符合2010年版药典标准。结论不同产地以及不同种植方式的甘草中,甘草酸、甘草苷及甘草总皂苷的含量差异较大,可为甘草规范化种植提供依据。     

Metabolic profiling of roots of liquorice (Glycyrrhiza glabra) from different geographical areas by ESI/MS/MS and determination of major metabolites by LC-ESI/MS and LC-ESI/MS/MS

Liquid chromatography electrospray mass spectrometry (LC-ESI/MS) has been applied to the full characterization of saponins and phenolics in hydroalcoholic extracts of roots of liquorice (Glycyrrhiza glabra). Relative quantitative analyses of the samples with respect to the phenolic constituents and to a group of saponins related to glycyrrhizic acid were performed using LC-ESI/MS. For the saponin constituents, full scan LC-MS/MS fragmentation of the protonated (positive ion mode) or deprotonated (negative ion mode) molecular species generated diagnostic fragment ions that provided information concerning the triterpene skeleton and the number and nature of the substituents. On the basis of the specific fragmentation of glycyrrhizic acid, an LC-MS/MS method was developed in order to quantify the analyte in the liquorice root samples. Chinese G. glabra roots contained the highest levels of glycyrrhizic acid, followed by those from Italy (Calabria).     

Comparative study of Glycyrrhiza glabra L. and Glycyrrhiza echinata L. of different origin. Medicinal plant polysaccharides III

Comparative study was carried out among the polysaccharides of Chinese, Lithuanian and Hungarian origin Glycyrrhiza glabra L. as well as the Hungarian origin Glycyrrhiza echinata L. Monosaccharide composition, measured as alditol acetates, of Chinese and Hungarian G. glabra was very similar, but the Lithuanian G. glabra and the G. echinata were quite different. The monosaccharides of polysaccharides isolated from root and stem of G. echinata show significant differences (Table II.). All investigated samples contain glucuronic acid, the G. echinata contains also galacturonic acid (Table II.). Some fractions with higher uronic acid content and the acidic polysaccharides isolated from these fractions were studied and their monosaccharide composition was determined (Table III. and IV.).     

Pharmaceutical botanical studies on some Glycyrrhiza species

Some Glycyrrhiza species grown in several domestic research gardens of medicinal plants were collected by the Osaka University of Pharmaceutical Sciences and were cultivated to compare their morphological properties. HPLC profile analysis was performed and index compounds of MeOH extracts of aerial parts and EtOAc extracts of subterranean parts were determined. Glycyrrhizin contents and growth rates of the underground parts of some types of Glycyrrhiza uralensis and Glycyrrhiza glabra were compared and four excellent types were selected as candidates for cultivation. One of them was due to Kanzo-Yashiki (Enzan, Yamanashi prefecture), where G. uralensis was cultivated in the Edo period. Alkaloidal constituents of G. uralensis and G. glabra were also investigated and anabasine (an insecticide) and a new tricyclic alkaloid were obtained.     

Species identification of licorice using nrDNA and cpDNA genetic markers

For the accurate identification of medicinal licorice species, nucleotide sequences of four types of DNA regions were researched for 205 specimens, including three species used as licorice: Glycyrrhiza uralensis, Glycyrrhiza glabra, and Glycyrrhiza inflata. The four DNA regions were the internal transcribed spacer (ITS) on nuclear ribosomal DNA, the rbcL gene, the matK gene, and the trnH-psbA intergenic region on chloroplast DNA (cpDNA). Ten genotypes were consequently recognized as combinations of the sequence data obtained from the four DNA regions. Species-specific genotypes were defined from the frequency of the appearance of species in each genotype and from the phylogenetic relationships of the 10 genotypes. This revealed the possibility of identifying licorice species based on the 10 genotypes. Next, comparison of species identifications by each DNA region suggested that efficient identification of licorice species is possible using the genetic information obtained from the ITS and trnH-psbA intergenic region. Additionally, concerning the phylogenetic relationships of the Glycyrrhiza species used as licorice, it is suggested from the genetic information of the four types of DNA regions that G. glabra is more closely related to G. inflata than to G. uralensis. In the G. uralensis examined, four genotypes were recognized as intra specific variations. The appearance frequency of each genotype in G. uralensis differed according to the area in China. G. uralensis may have expanded its distribution areas from western to eastern China because many licorices with the phylogenetic ancestral genotype were observed in western areas, while many with the derivative genotype were observed in eastern areas.     

云南甘草总皂甙清除氧自由基和抗脂质过氧化作用

云南甘草总皂甙(SGY)对Fenton反应生成的·OH具有较强的直接清除作用,其IC_(50)为29mg·L~(-1).且作用明显强于·OH特异性清除剂甘露醇(其IC_(50)为2.27×10~(-2)mol·L~(-1).即4315mg·L~(-1))。SGY对黄嘌呤-黄嘌呤氧化酶系统产生的O_2~+也有较强的清除作用·其IC_(50)为89mg·L~(-1)SGY对自由基发生系统(FRGS)诱导离体大鼠心肌匀浆组织产生的脂质过氧化(LPO)作用有明显的抑制效应.对FRGS诱导离体大鼠心肌线粒体膜LPO也有一定的抑制作用。结果提示SGY的抗脂质过氧化作用与其清除氧自由基作用有关。     

甘草对大鼠离体输精管的影响

去甲肾上腺素、乙酰胆碱、组胺均可引起大鼠离体输精管的收缩，此作用可被甘草提取液（667μg／ml）所拮抗，提示甘草的解痉作用可能与α，M和H受体有关。     

Protection of mitochondrial functions against oxidative stresses by isoflavans from Glycyrrhiza glabra

Isoflavan derivatives, glabridin (1), hispaglabridin A (2), hispaglabridin B (3), 4'-Omethylglabridin (4) and 3'-hydroxy-4'-O-methylglabridin (5), isolated from Glycyrrhiza glabra, were investigated for their ability to protect liver mitochondria against oxidative stresses. Mitochondrial lipid peroxidation linked to respiratory electron transport and that induced non-enzymatically were inhibited by these isoflavans. Hispaglabridin A (2) strongly inhibited both peroxidations and 3'-hydroxy-4'-O-methylglabridin (5) was the most effective at preventing NADH-dependent peroxidation. 3'-Hydroxy-4'-O-methylglabridin (5) protected mitochondrial respiratory enzyme activities against NADPH-dependent peroxidation injury. Dihydroxyfumarate-induced mitochondrial peroxidation was also prevented by this isoflavan. Isoflavans from G. glabra were shown to be effective in protecting mitochondrial function against oxidative stresses.     

新疆胀果甘草化学成分的分离与鉴定

目的研究中药胀果甘草(Glycyrrhiza inflataBatal.)生产甘草酸后的工业尾料的化学成分,为进一步合理利用这一传统中药资源提供依据。方法采用反复硅胶柱色谱、聚酰胺柱色谱、Sepha-dex LH-20凝胶柱色谱等方法进行分离纯化,并通过理化常数测定与光谱分析鉴定其化学结构。结果从胀果甘草药渣的体积分数95%乙醇提取物中分离鉴定了9个化合物,分别为甘草查耳酮A(licochalcone A,1)、2′,4,4′-三羟基查耳酮(2′,4,4′-trihydroxychalcone,2)、(-)α,2′,4,4′-四羟基二氢查耳酮((-)α,2′,4,4′-tetrahydroxydihydrochalcone,3)、甘草查耳酮C(licochalcone C,4)、甘草查耳酮D(licochalcone D,5)、刺甘草查耳酮(echinatin,6)、kanzonol E(7)、咖啡酸二十二酯(docosylcaffeate,8)、甘草次酸(glycyrrhetinic acid,9)。结论其中化合物2~5、7~9均为首次从甘草药渣中分离得到,化合物2、3、7、8均为首次从胀果甘草中分离得到。     

Constituent properties of licorices derived from Glycyrrhiza uralensis, G. glabra, or G. inflata identified by genetic information

Constituent properties of licorices derived from Glycyrrhiza uralensis, G. glabra, and G. inflata are revealed by comparing 117 of licorice identified using four genetic markers; internal tracscribed spacer (ITS) on nuclear ribosomal DNA, rbcL gene, matK gene, and trnH-trnK1 intergenic region on chloroplast DNA. Regarding six main constituents of licorice; glycyrrhizin, liquiritin, liquiritin apioside, isoliquiritin, isoliquiritin apioside, and liquiritigenin, the constituent property of G. glabra resembles to that of G. inflata. On the other hand, the constituent property of G. uralensis is not similar to that of G. glabra or G. inflata and is characterized by a wide content variation of the six constituents compared to those of G. glabra and/or G. inflata. The mean contents of liquiritin, isoliquiritin, or liquilitigenin in G. uralensis are significantly higher than those of G. glabra or G. inflata. Therefore, the licorice species should be selected depending on these constituent properties for the traditional Chinese medicines or the Japanese Kampo medicines. Additionally, glycycoumarin, glabridin, and licochalcone A were reconfirmed as the species-specific typical constituents of G. uralensis, G. glabra, and G. inflata respectively. Therefore, it is resulted that the determination of the three species-specific constituents may be useful for the species identification of licorice. However, since 6% of licorice examined and hybrids were exceptions to the rule, their genetic information is necessary for the accurate species identification of licorice.     

药用甘草离体快速繁殖体系的建立

筛选合适的甘草快速繁殖技术体系,为甘草的大规模生产生物活性物质、基因工程和人工制种奠定基础。以乌拉尔甘草、黄甘草和光果甘草为供试材料,以其子叶茎段和侧芽茎段作外植体,对影响甘草快速繁殖形成和发生的因素进行研究。结果显示乌拉尔甘草诱导的丛生苗数较多,是3个供试材料中最好的;甘草适宜的外植体是子叶茎段。利用甘草子叶茎段和侧芽茎段作外植体进行快速繁殖,具有生长周期短,再生频率高,适于规模化工厂生产。     

Phenolic constituents of licorice. IV. Correlation of phenolic constituents and licorice specimens from various sources, and inhibitory effects of licorice extracts on xanthine oxidase and monoamine oxidase

The roots and/or rhizomes of Glychyrrhiza uralensis, G. glabra and G. inflata, and commercial licorice specimens from various regions or countries were analyzed by high-performance liquid chromatography (HPLC), and classified into three types based on their phenolic constituents. i) Type A: The roots and rhizomes of G. uralensis, commercial licorice specimens from northwestern region of China (Seihoku-kanzo) and from northeastern region of China (Tohoku-kanzo) in Japanese markets, and also several licorice specimens from Chinese markets. They contain licopyranocoumarin (6), glycycoumarin (7) and/or licocoumarone (8), which were not found in G. glabra and G. inflata. ii) Type B: The root and rhizome of G. glabra, and the licorice specimens imported from the Soviet Union and Afghanistan. They contain glabridin (9) and glabrene (10), which were not found in the samples of the other two Glycyrrhiza species. A root sample of Glycyrrhiza species from Turkey also contains 9 and 10. iii) Type C: The root sample of G. inflata. They contain licochalcones A (11) and B (12), which were not found in the samples of the other two Glycyrrhiza species. Commercial licorice specimens obtained in Japan, which were imported from Sinkiang of China (Shinkyo-kanzo), and some licorice specimens obtained from Chinese markets, have also been found to contain 11 and 12. The phenolics 6-12, characteristic constituents of types A, B or C, were not found in a specimen of cortex-free licorice from a Japanese market (kawasari-kanzo). Extracts of some licorice specimens of types A and B, and all of the licorice specimens of type C inhibited 40-56% of the xanthine oxidase activity at the concentration of 30 micrograms/ml. Extracts of some licorice specimens of types A and B also showed inhibitory effects on monoamine oxidase (44-64% inhibition, at the concentration of 30 micrograms/ml), which were slightly weaker than that of harmane hydrochloride.     

芍药甘草汤中自组装纳米粒形成对甘草主要成分体外释放和肠吸收的影响

目的 研究芍药甘草汤中自组装纳米粒(SGD-SAN)形成对甘草主要成分体外释放和肠吸收的影响.方法 制备甘草单煎液(GSD)、芍药单煎液(SSD)、芍药-甘草单煎混合液(MSSGD)和SGD(即芍药、甘草合煎液),并对其中的SAN进行表征.采用高效液相色谱法测定甘草中7种主要成分(芹糖甘草苷、甘草苷、芹糖异甘草苷、异甘...     

Two minor chalcone acetylglycosides from the roots extract of <Emphasis Type="Italic">Glycyrrhiza uralensis</Emphasis>

An extensive phytochemical investigation on the roots of Glycyrrhiza uralensis led to the isolation of two new minor chalcone acetylglycosides, i.e., 6″-O-acetylisoliquiritin (1) and 6″-O-acetylneoisoliquiritin (2), including 16 kinds of known constituents (3–18) of flavonoids, chalcones and triterpene saponins. The chemical structures of 1 and 2 were established by spectroscopic analyses of them, particularly by the aid of two-dimensional NMR experiments, COSY, DEPT, HSQC and HMBC. Some isolated components except 1 and 2 exhibited significant inhibitory effects on the proliferation of cultured tumor cell lines, such as A549, SK-OV-3, A-498, and HCD15, in vitro.