中药大蓟化学成分的研究

从大蓟的50％乙醇提取物中分离得到2个木脂素：（-）2-（3’-甲氧基4’-羟基-苯基）-3,4-二羟基4-（3＂-4＂-羟基-苄基）-3-四氢呋哺甲醇（1）和络石苷（2）,以及另外6个化合物：蒙花苷（3）、柳穿鱼叶苷（4）、粗毛豚草素（5）、芹菜素（6）、咖啡酸（7）和对-香豆酸（8）。本文首次在蓟属植物中发现木脂素类成分,化合物7也为首次从本植物中分离得到,通过体外玻片法对化合物1—8进行凝血活性测定,发现化合物3、4具有一定的促凝血作用。     

Linarin and its aglycone acacetin abrogate actin ring formation and focal contact to bone matrix of bone-resorbing osteoclasts through inhibition of αvβ3 integrin and core-linked CD44

BackgroundSince enhanced bone resorption due to osteoclast differentiation and activation cause skeletal diseases, there is a growing need in therapeutics for combating bone-resorbing osteoclasts. Botanical antioxidants are being increasingly investigated for their health-promoting effects on bone. Edible Cirsium setidens contains various polyphenols of linarin, pectolinarin, and apigenin with antioxidant and hepatoprotective effects.PurposeThis study aimed to determine whether linarin present in Cirsium setidens water extracts (CSE) and its aglycone acacetin inhibited osteoclastogenesis of RANKL-exposed RAW 264.7 murine macrophages for 5 days.MethodsThis study assessed the osteoprotective effects of CSE, linarin and acacetin on RANKL-induced differentiation and activation of osteoclasts by using MTT assay, TRAP staining, Western blot analysis, bone resorption assay actin ring staining, adhesion assay and immunocytochemical assay. This study explored the underlying mechanisms of their osteoprotection, and identified major components present in CSE by HPLC analysis.ResultsLinarin and pectolinarin were identified as major components of CSE. Nontoxic linarin and acacetin as well as CSE, but not pectolinarin attenuated the RANKL-induced macrophage differentiation into multinucleated osteoclasts, and curtailed osteoclastic bone resorption through reducing lacunar acidification and bone matrix degradation in the osteoclast-bone interface. Linarin and acacetin in CSE reduced the transmigration and focal contact of osteoclasts to bone matrix-mimicking RGD peptide. Such reduction was accomplished by inhibiting the induction of integrins, integrin-associated proteins of paxillin and gelsolin, cdc42 and CD44 involved in the formation of actin rings. The inhibition of integrin-mediated actin ring formation by linarin and acacetin entailed the disruption of TRAF6-c-Src-PI3K signaling of bone-resorbing osteoclasts. The functional inhibition of c-Src was involved in the loss of F-actin-enriched podosome core protein cortactin-mediated actin assembly due to linarin and acacetin.ConclusionThese observations demonstrate that CSE, linarin and acacetin were effective in retarding osteoclast function of focal adhesion to bone matrix and active bone resorption via inhibition of diffuse cloud-associated αvβ3 integrin and core-linked CD44.     

A morphological, karyological and chemical study of the Apteranthes (Caralluma) europaea complex

The Apteranthes europaea complex (Apocynaceae–Asclepiadoideae–Ceropegieae–Stapeliinae) distributed from Morocco/southern Spain along the North African coast of the Mediterranean Sea up to Sinai, Negev and southern Jordan is studied with emphasis on stem and flower morphology by SEM, chromosome sizes and flavonoid compounds. Apteranthes europaea was found to be rich in rather uncommon flavone glycosides. Of these, luteolin 4'-neohesperidoside represents the major flavonoid of all samples; luteolin-3'- O -(6'- O -sinapoylglucoside)-4'- O -neohesperidoside and luteolin-3'- O -(6'- O -feruloylglucoside)-4'- O -neohesperidoside are reported here for the first time. Flavonol glycosides also occur, but in much smaller quantities. The different flower morphs, variation in stem and corolla epidermal structures, slight variations in the length of the 2 n  = 22 chromosomes and in quantitative flavonoid composition are taxonomically best reflected by subdividing the complex into a western var. europaea (Europe and Africa) and an eastern var. judaica (Sinai, Arabia).  © 2005 The Linnean Society of London, Botanical Journal of the Linnean Society , 2005, 149 , 419–432.     

蓟属两种植物的染色体研究

本文对蓟属（Ｃｉｒｓｉｕｍ）的两个形态相似的近缘种大刺儿菜和小刺儿菜进行了染色体研究,其中后者为首次报道。观察结果表明：两个种的染色体数目均为２ｎ＝２ｘ＝３４：它们的核型是：大刺儿菜．２ｎ＝２ｘ＝３４＝２０ｍ＋１２ｓｍ＋２ｓｔ：小刺儿菜．２ｎ＝２ｘ＝３４＝２２ｍ＋１０ｋｍ（２ＳＡＴ）＋２ｓｔ。通过核型比较,认为它们是两个独立的种．而且后者比前者进化。     

蓟属二种植物过氧化物同工酶的酶谱式样及其分类学意义

采用聚丙烯酰胺凝胶电泳的方法对蓟属的两个近缘种：大刺儿菜和小刺儿菜的过氧化物酶同工酶分别在不同的发育阶段和不同的生长下进行了比较研究。结果表明,二者的酶谱存在 明显的差异,从而支持将它们作为两个的种对待。此外,本文还对ＰＯＸ酶谱的分类学意义进行了探讨。     

Two new flavone glycosides from Cirsium lineare

An examination of four species of Cirsium disclosed the presence of two new flavonoids in C. lineare. The structure of one was 5,4′-dihydroxy-6,7,3′-trimethoxyflavone (cirsilineol) 4′-monoglucoside and the other 5,3′,4′-trihydroxy-6,7-dimethoxyflavone (cirsiliol) 4′-monoglucoside. Luteolin 7-glucoside was found in C. suffultum, and pectolinarin and linarin in C. kamtschaticum and C. pectinellum.     

刺儿菜复合体的形态学研究

本文对不同产地,不同居群的全国２００份刺儿菜复合体标本的若干性状选用Ｅ．Ａｎｄｅｒｓｏｎ的形象化散点图和柱状图方法进行研究分析,认为将大刺儿菜与小刺儿菜分为独立的种是合理的。     

Leaf traits of congeneric host plants explain differences in performance of a specialist herbivore

1. Within the host range of herbivorous insects, performance hierarchies are often correlated with relatedness to a primary host plant, as plant traits are phylogenetically conserved. Therefore, it was hypothesised that differences in herbivore performance on closely related plant species are due to resistance traits that vary in magnitude, rather than in the nature of the traits. 2. This hypothesis was tested by manipulating putative resistance traits of three congeneric thistle species (Cirsium arvense, Cirsium palustre, and Cirsium vulgare) and assessing the performance of the oligophagous, leaf‐feeding beetle, Cassida rubiginosa. Measurements were done of survival, weight gain, and development time of the beetle on its primary host, C. arvense, and two alternative hosts under low and high nutrient availability, and on shaved and unshaved leaves. 3. Survival of C. rubiginosa was strongly dependent on plant species with final mean survival rates of 47%, 16%, and 8% on C. arvense, C. palustre, and C. vulgare, respectively. Survival was primarily explained by leaf trichome densities, and to a lesser extent by specific leaf area. Leaf flavonoid concentrations did not explain differences in beetle survival, and there were no differences in beetle weight gain or development time of individuals that survived to adulthood. 4. No beetles survived on unshaved (hairy) C. vulgare plants, but manipulating leaf trichome densities of the thistle species by shaving the leaves moderated the plant‐specific resistance, and equalised the survival rates. Survival of C. rubiginosa on alternative congeneric hosts was explained by a common physical resistance trait that varied in magnitude.     

Phenolic Glycosides with antiproteasomal activity from Centaurea urvillei DC. subsp. urvillei

A new flavanone glycoside, naringenin-7-O-β-d-glucuronopyranoside, and a new flavonol glycoside, 6-hydroxykaempferol-7-O-β-d-glucuronopyranoside were isolated together with 12 known compounds, 5 flavone glycoside; hispidulin-7-O-β-d-glucuronopyranoside, apigenin-7-O-β-d-methylglucuronopyranoside, hispidulin-7-O-β-d-methylglucuronopyranoside, hispidulin-7-O-β-d-glucopyranoside, apigenin-7-O-β-d-glucopyranoside, a flavonol; kaempferol, two flavone; apigenin, and luteolin, a flavanone glycoside; eriodictyol-7-O-β-d-glucuronopyranoside, and three phenol glycoside; arbutin, salidroside, and 3,5-dihydroxyphenethyl alcohol-3-O-β-d-glucopyranoside from Centaurea urvillei subsp. urvillei. The structure elucidation of the new compounds was achieved by a combination of one- (¹H and ¹³C) and two-dimensional NMR techniques (G-COSY, G-HMQC, and G-HMBC) and LC-ESI-MS. The isolated compounds were tested for their antiproteasomal activity. The results indicated that kaempferol, a well known and widely distributed flavonoid in the plant kingdom, was the most active antiproteasomal agent, followed by apigenin, eriodictyol-7-O-β-d-glucuronopyranoside, 3,5-dihydroxyphenethyl alcohol-3-O-β-d-glucopyranoside, and salidroside, respectively.     

Chemotaxonomic consideration of flavonoids from the leaves of Chrysanthemum arcticum subsp. arcticum and yezoense,and related species

We examined the foliar flavonoids of Chrysanthemum arcticum subsp. arcticum and yezoense, and related Chrysanthemum species. Five flavonoid glycosides (luteolin 7-O-glucoside and 7-O-glucuronides of luteolin, apigenin, eriodictyol and naringenin) were isolated from these taxa. Luteolin 7-O-xylosylglucoside, luteolin, apigenin and quercetin 3-methyl ether were found in subsp. yezoense as very minor compounds that were not recognised by high-performance liquid chromatography/photodiode array (HPLC/PDA). The related species C. yezoense contained acacetin 7-O-rutinoside and some methoxylated flavone aglycones as major compounds. Thus, C. arcticum was distinguished from C. yezoense according to their flavonoid profiles.     

蕨菜黄酮苷元的组成及含量测定研究

以80%乙醇浸提经乙醚脱脂预处理的干蕨菜粉,减压回收乙醇得浓缩液.浓缩液以8%硫酸沸水浴水解3 h后,加入20%NaOH中和至弱酸性,过滤,滤渣以三氯甲烷索氏提取获总蕨菜黄酮苷元.TLC法检测总黄酮苷元的组成,HPLC法测定其主要苷元的含量.结果表明,通过酸水解获得总蕨菜黄酮苷元,其主要成分为山奈酚,含量约占蕨菜干重的1.9%.     

Purification,structural elucidation,antioxidant capacity and neuroprotective potential of the main polyphenolic compounds contained in Achyrocline satureioides (Lam) D.C. (Compositae)

Achyrocline satureioides (Lam) D.C (Compositae) is a native medicinal plant of South America traditionally utilized for its anti-inflammatory, sedative and anti-atherosclerotic properties among others. Neuroprotective effects have been reported in vivo and could be associated to its elevated content of flavonoid aglycones. In the present study we performed the isolation and structure elucidation of the major individual flavonoids of A. satureioides along with the in vitro characterization of their individual antioxidant and neuroprotective properties in order to see their putative relevance for treating neurodegeneration.Exact mass, HPLC-MS/MS and ¹H NMR identified dicaffeoyl quinic acid isomers, quercetin, luteolin, isoquercitrin, and 3-O-methylquercetin as the mayor polyphenols. Flavonoids intrinsic redox properties were evaluated in the presence of the endogenous antioxidants GSH and Ascorbate. Density Functional Theory (DFT) molecular modeling and electron density studies showed a theoretical basis for their different redox properties. Finally, in vitro neuroprotective effect of each isolated flavonoid was evaluated against hydrogen peroxide-induced toxicity in a primary neuronal culture paradigm. Our results showed that quercetin was more efficacious than luteolin and isoquercitrin, while 3-O-methylquercetin was unable to afford neuroprotection significantly. This was in accordance with the susceptibility of each flavonoid to be oxidized and to react with GSH. Overall our results shed light on chemical and molecular mechanisms underlying bioactive actions of A. satureioides main flavonoids that could contribute to its neuroprotective effects and support the positive association between the consumption of A. satureioides as a natural dietary source of polyphenols, and beneficial health effect.     

一株高抗氧化活性内生真菌的分子鉴定及产酚酸类物质研究

【目的】以疏花水柏枝(Myricaria laxiflora)淹水前根部分离到的内生真菌SG17为研究对象,为明确其抗氧化能力及可能的物质基础,对其进行生物学鉴定、抗氧化能力分析及产酚酸类物质研究。【方法】对该菌nr DNA的内转录间隔区进行测序;应用总抗氧化试剂盒,以维生素C为阳性对照,测定其抗氧化能力;采用高效液相色谱(HPLC)和薄层色谱(TLC)探讨其发酵液中的酚酸类化合物。【结果】SG17具有很高的抗氧化活性,其发酵液粗提物的抗氧化活性达到同浓度抗坏血酸的31.86%,分子和形态鉴定表明SG17是一株烟曲霉(Aspergillus fumigatus)。其发酵产物中酚酸类物质含量丰富,HPLC能检测到含有没食子酸,且通过HPLC和TLC分析还发现一种光学性质与绿原酸极为相似的类似物。【结论】内生真菌烟曲霉SG17发酵液的抗氧化能力远高于国际报道水平,是一种具有潜在开发价值的抗氧化剂资源。     

Free radical scavengers from Cymbopogon citratus (DC.) stapf plants cultivated in bioreactors by the temporary immersion (TIS) principle

The biomass production of Cymbopogon citratus shoots cultivated in bioreactors according to the temporary immersion (TIS) principle was assessed under different growth conditions. The effect of gassing with CO2-enriched air, reduced immersion frequency, vessel size and culture time on total phenolic and flavonoid content and free radical scavenging effect of the methanolic extracts was measured. From the TIS-culture of C. citratus, seven compounds were isolated and identified as caffeic acid (1), chlorogenic acid (2), neochlorogenic acid (3), p-hydroxybenzoic acid (4), p-hydroxybenzoic acid 3-O-beta-D-glucoside (5), glutamic acid (6) and luteolin 6-C-fucopyranoside (7). The occurrence of compounds 1-7 and their variability in C. citratus grown under different TIS conditions was determined by HPLC. The free radical scavenging effect of the methanolic extract and compounds was measured by the discoloration of the free radical 1,1-diphenyl-2-picrylhydrazyl (DPPH). The main metabolites in 6- and 8-week-old cultures, both in 5 and 10 1 vessels, were chlorogenic acid (2) (100-113 mg%) and neochlorogenic acid (3) (80-119 mg%), while in the cultures with CO2-enriched air and reduced immersion frequency the main compound detected in the extracts was glutamic acid (6) (400 and 670 mg% for the green and white biomass and 619 and 630 mg% for the green and white biomass, respectively). The most active compounds, as free radical scavengers, in the DPPH discoloration assay were caffeic acid (1), chlorogenic acid (2), neochlorogenic acid (3) and the flavonoid luteolin 6-C-fucopyranoside (7).     

Development of polymorphic markers for Cirsium arvense,Canada thistle,and their amplification in closely related taxa

Suppression of invasive Canada thistle, Cirsium arvense, with biological control agents has stalled because introduced agents were not host‐specific. To aid in the development of more effective management strategies, molecular markers are needed to examine the genetic structure of Canada thistle populations. Microsatellite (simple sequence repeat) markers were developed and intersimple sequence repeat (ISSR) markers were tested for North American populations. An average of nine polymorphic alleles per microsatellite locus and 11 per ISSR locus were detected. These will be used to examine the genetic structure of C. arvense in the northern Great Plains and their transferability to endemic Cirsium spp.     

A Comparison of Auxotrophic and Wild Strains of Sclerotinia sclerotiorum Used as a Mycoherbicide Against Californian Thistle ( Cirsium arvense )

Two auxotrophic mutant strains of Sclerotinia sclerotiorum were tested in the greenhouse for pathogenicity on Cirsium arvense (Californian thistle) with and without amino acid amendments. An arginine auxotrophic mutant, with an amendment of the amino acid, followed an identical disease progress curve to that of the wild strain of the pathogen from which it was derived. However, when deprived of the amino acid amendment it was still highly pathogenic. A leucine auxotrophic mutant demonstrated poor pathogenicity without a leucine amendment, but improved pathogenicity with the addition of the amino acid. However, both of these treatments were inferior to the two wild strains tested and the arginine auxotroph with and without amendments. A field experiment was conducted on C. arvense stems in permanent pasture to compare the pathogenicity of amended auxotrophic strains and wild strains of S. sclerotiorum applied as a granule in a wheat-based carrier. The two wild strains gave significant reductions in thistle cover within 3 months of treatment, and subsequent reductions in thistle stem height and density during the following season. There was no evidence that the auxotrophic strains reduced thistle cover in the season the treatments were applied, but they did reduce subsequent stem density in the following spring. To determine disease carry-over associated with the wild and auxotrophic strains of the pathogen, rape was planted into subplots over the next three consecutive seasons. Despite substantial populations of sclerotia being present in the soil, especially in the first season after treatment of the thistles, no disease of rape caused by S. sclerotiorum was detected over the three seasons in any of the plots. Sclerotium populations of S. sclerotiorum in the soil declined by over 50% between 20 and 32 months after treatment, but there was no decline over the subsequent 12 months. The trial demonstrated that the auxotrophic strains were less field fit compared with the wild strains and that the presence of inoculum and a susceptible host to S. sclerotiorum were not the only prerequisites for disease development. It was concluded that use of a trap crop following treatment is not a suitable method for determining the risk of using this pathogen as a mycoherbicide in pasture.     

高速逆流色谱法分离凤尾草中的芹菜素和木犀草素

建立了高速逆流色谱分离纯化芹菜素和木犀草素的方法。两相溶剂系统为氯仿-甲醇-水（4：3：2）,上相为固定相,下相为流动相进行洗脱。从100mg粗提物中一步分离得到17．0mg芹菜素和22．5mg木犀草素,经高效液相色谱分析,纯度分别为92％和96％。其化学结构由1H和13CNMR鉴定。     

Application of high-performance liquid chromatography to the determination of vinblastine in Catharanthus roseus

A method for the determination of vinblastine in Catharanthus roseus by HPLC is described. A crude alkaloid extract, obtained by extraction of leaves with toluene, 2% citric or tartaric acid and benzene, was separated by TLC. The vinblastine fraction was cut out and vinblastine was eluted from the sorbent. The amount of vinblastine was determined by HPLC with peak-height measurement. The standard deviation is 0.2 μg/ml. The detection limit is 0.05 ng of vinblastine in a sample applied to the HPLC column.     

Chemosystematic investigations on phenolics from flowerheads of Central European taxa of Hieracium sensu lato (Asteraceae)

 HPLC-UV and HPLC-MS investigations of phenolic acids and flavonoids in flowerheads of 84 samples of 76 taxa belonging to 66 species of Hieracium resulted in the identification of three phenolic acids (chlorogenic acid, 3,5-dicaffeoyl quinic acid, 4,5-dicaffeoyl quinic acid) and six flavonoids (apigenin 4^′-O-β-D-glucuronide, isoetin 4^′-O-β-D-glucuronide, luteolin, luteolin 7-O-β-D-glucoside, luteolin 7-O-β-D-glucuronide, luteolin 4^′-O-β-D-glucoside). The contents of these secondary metabolites were quantified by HPLC using quercetin and cynarin as internal standards. In contrast to the previously investigated genera Leontodon and Crepis, cichoric acid and caffeoyl tartaric acid were not found in any of the investigated Hieracium taxa. Results of HPLC analyses revealed only a limited degree of qualitative variation between the different taxa, and luteolin 7-O-β-D-glucuronide and isoetin 4^′-O-β-D-glucuronide were the only compounds, which were not detectable in some of the investigated taxa. Quantitative patterns of phenolics differed markedly between particular taxa and Principal Component Analysis of the quantification results yielded separate clusters for the members of the subgenera Hieracium and Pilosella. Received January 23, 2001 Accepted October 11, 2001