淡紫松果菊生药学研究

为淡紫松果菊的鉴别与开发利用提供依据。方法 形态、性状与显微鉴定。结果 淡紫松果菊与狭叶松果菊虽然内部结构极为相似，但二者有一些明显区别点。结论 研究结果可为制订淡紫松果菊的质量标准提供依据。     

松果菊属3种植物的理化分析

对松果菊属紫花松果菊、狭叶松果菊,淡紫松果菊3个种进行了理化分析研究。方法 采用HPLC、TLC法对3种松果菊的松果菊苷进行了定性和定量分析;采用紫外扫描仪,对3种松果菊进行了紫外吸收光谱的测定。结果 3种松果菊根和茎叶中均含松果菊苷成分,但紫花松果菊的含量最低。结论 通过理化分析表明,3种松果菊之间的成分组成存在着差异,特别是紫花松果菊根与另两种根之间存在显著差异。该结果为进一步合理开发利用松果菊资源提供了参考资料。     

3种松果菊属植物的鉴别、活性成分及生物技术研究进展

综述了近年来关于3种药用松果菊:狭叶松果菊E ch inacea angustif olia、紫花松果菊E.purpurea和淡紫松果菊E.p a llid a的生物活性成分和生物技术方面的研究进展以及这3种药用松果菊的鉴别方法,并对松果菊属植物的药理活性进行了总结。药用松果菊的主要活性成分是多糖、糖蛋白、咖啡酸衍生物和烷基酰胺类。松果菊制剂具有增强免疫、抗炎、抗感染等功效。在生物技术领域,有关松果菊属植物的首例转基因已在狭叶松果菊中取得成功。采用离体培养法已建立了松果菊的植株再生体系,实现了植株的离体快繁。     

狭叶松果菊的形态、性状与显微鉴别研究

目的 为狭叶松果菊鉴别与开发利用提供依据。方法 形态、性状与显微鉴定。结果 完成了原植物形态、药材性状与显微特征的描述。结论 部分结果可为制订狭叶松果菊的质量标准提供依据。     

狭叶松果菊的毛状根诱导及培养

目的 利用两种发根农杆菌A4,R1000诱导狭叶松果菊产生毛状根,建立狭叶松果菊的毛状根培养体系.方法 利用共培养法研究不同外植体、菌株、预培养时间和浸染时间等对狭叶松果菊毛状根诱导率的影响和不同液体培养基对毛状根生长的影响筛选出最佳培养基.结果 利用发根农杆菌R1000,预培养48 h的叶柄为转化材料,浸染10 min的诱导率最高,毛状根悬浮培养的最佳培养基为1/2MS+ IBA0.5液体培养基.结论 狭叶松果菊毛状根离体培养体系的建立,为狭叶松果菊的次生代谢产物的大规模生产奠定了基础.     

狭叶松果菊组织培养不定芽发生与继代增殖

狭叶松果菊Echinacea angustifolia为菊科松果菊属的3个主要种之一,原产于美洲,是印地安民间的传统药材,被用作治疗外伤、蛇咬、头痛及感冒等,具有较强的免疫促进和调节活性[1]。狭叶松果菊较早在我国北京怀柔进行引种栽培,其他地区也有少量的引种栽培。狭叶松果菊可全草入药,主要成分为多糖和糖蛋白、咖啡酸衍生物以及烷基酰胺类、不饱和酮类、倍半萜类和生物碱类等化合物[2,3]。近年来,由于松果菊属植物具有重要的药用价值,其....     

三种松果菊化学成分与生物活性研究进展

松果菊是菊科Compositae松果菊属Echinacea植物,分布在北美洲,该属植物约有9种[1] ,主要药用种是狭叶松果菊E .angustifolia ,紫花松果菊E .purpurea和淡紫松果菊E .pallida。松果菊的治疗用途十分广泛,可以治疗各种炎症和感染,其疗效得到了越来越多的临床医生的肯定[1～4] 。对松果菊的研究也日趋深入,有关其化学成分和药理活性方面的研究已经进行了大量的工作,有了比较深入的认识,现将其化学和药理方面的研究综述如下。1　化     

DNA分子标记技术在龙胆鉴别与亲缘关系划分上的应用

研究东北产药材龙胆的鉴别方法及其亲缘关系，用分子生物学技术对东北产药材龙胆的3个不同品种进行了DNA提取和RAPD及ISSR分析。应用RAPD法从80个随机引物中找出了3种引物（S-76，S-69，S-64）来标记3种龙胆发现这3种引物能把供试的3种龙胆全部区分开。同时应用ISSR法从7个引物中筛选出4个引物能扩增出稳定遗传多态性引物，共扩增出44条DNA条片段，其中同源片段有22个，特异片段有12个，多态片段为10个。用这些引物扩增出的指纹图谱进行聚类分析，得出了相同的结论，即在3种龙胆中，条叶龙胆，龙胆亲缘关系较近，三花龙胆与二者亲缘关系较远。结果表明分子标记技术可用于龙胆的鉴别，亲缘关系划分。     

狭叶松果菊3-脱氢奎尼酸合成酶基因cDNA的克隆及其组织表达特征

目的 克隆狭叶松果菊Echinacea angustifolia 3-脱氢奎尼酸合成酶基因并考察其在各个组织中的表达情况.方法 采用快速扩增cDNA末端技术,以狭叶松果菊组培苗cDNA为模板,克隆出狭叶松果菊3-脱氢奎尼酸合成酶基因全长序列并通过半定量RT～PCR分析其在不同器官中的表达模式.结果 克隆到的基因(命名为EanaroB)全长为1 424 bp,编码一个442个氨基酸残基组成的多肽.其氨基酸序列与植物来源的3-脱氢奎尼酸合成酶同源性都在80%左右.将得到的序列提交Genbank,序列号为EU293857.半定量RT-PCR结果 表明,狭叶松果菊EanaroB基因在狭叶松果菊的根、茎、叶、花中均有表达,但花和叶中的表达量较高,在根和茎中较少.结论 采用RACE和PCR的方法 从狭叶松果菊中克隆出了咖啡酸类化合物生物合成途径上的一个基因EanaroB,为进一步研究咖啡酸类衍生物生物合成代谢途径提供一定依据,为今后利用基因工程技术提高咖啡酸类衍生物,包括苯乙醇苷类物质松果菊苷的代谢工程打下一定基础.     

柴胡与狭叶柴胡的特异性引物PCR鉴别方法

目的设计专用于柴胡与狭叶柴胡鉴别的ARMS特异性引物,为柴胡、狭叶柴胡的药材鉴别提供分子鉴定依据。方法根据柴胡、狭叶柴胡与其他柴胡属植物的r DNA ITS序列,寻找柴胡与狭叶柴胡的序列特异性位点,设计用于进行柴胡与狭叶柴胡分子鉴别的特异性引物。结果该特异性引物为柴胡、狭叶柴胡的鉴别提供了分子鉴定依据,可从多种药材及中成药(蜜丸、水蜜丸)中快速、准确地检测出柴胡、狭叶柴胡。结论运用特异性引物PCR法能有效地从植物和中成药(蜜丸及水丸)中鉴别出柴胡与狭叶柴胡,该方法具有准确、高效、省时、简便的特点。     

Anti-inflammatory and cicatrizing activity of Echinacea pallida Nutt. root extract.

Among the different species belonging to the Echinacea family, largely used in traditional medicine, Echinacea pallida, Echinacea purpurea and Echinacea angustifolia were investigated. These different species, due to their difficult identification, were commonly confused in the past and probably used indifferently for the same therapeutic purposes. In fact, the three species have in common, some pharmacological activities, based on the presence of active compounds that act additively and synergistically. Nevertheless, the composition of each species has slight variation in the amount of each active component. In particular, echinacoside, a caffeoyl derivative, is present in E. pallida and only in traces in E. angustifolia. It seems to have protective effects on skin connective tissue and to enhance wound healing. The anti-inflammatory and wound healing activities of echinacoside, compared with the ones of the total root extract of E. pallida and E. angustifolia, were examined in rats, after topical application. The tissues of the treated animals were evaluated after 24, 48 and 72 h treatment and excised for histological observation at the end of the experiment. Results confirm the good anti-inflammatory and wound healing properties of E. pallida and of its constituent echinacoside, with respect to E. purpurea and control. This activity probably resides in the antihyaluronidase activity of echinacoside.     

Cytotoxic effects of Echinacea root hexanic extracts on human cancer cell lines

Echinacea is one of the most widely used alternative medicine in the world. Intake of Echinacea preparations is common among patients with advanced malignancies enrolled onto phase I chemotherapy trials; however, to our knowledge, no data are available regarding the possible direct effect of Echinacea species on human cancer cells. The purpose of the present study was to investigate potential in vitro cytotoxic and pro-apoptotic properties of hexanic root extract of the three medicinal Echinacea (Asteraceae) species (Echinacea pallida (Nutt.) Nutt., Echinacea angustifolia DC. var. angustifolia, Echinacea purpurea (L.) Moench.) on the human pancreatic cancer MIA PaCa-2 and colon cancer COLO320 cell lines. We demonstrated, for the first time, that all the three species reduced cell viability in a concentration- and time-dependent manner; Echinacea pallida was the most active species with IC(50)s of 46.41+/-0.87 and 10.55+/-0.70 microg/ml in MIA PaCa-2 and COLO320 cells, respectively. Echinacea pallida extract was able to induce apoptosis by increasing significantly caspase 3/7 activity and promoting nuclear DNA fragmentation. These results represent the starting point to establish viable scientific evidence on the possible role of Echinacea species in medical oncology.     

Echinacea purpurea and P-glycoprotein drug transport in Caco-2 cells

Echinacea is widely used as a medical herbal product, but its interaction potential with the drug efflux transporter P-glycoprotein (P-gp) has not yet been evaluated. The interaction potential of Echinacea purpurea towards P-gp mediated drug transport was studied in human intestinal Caco-2 cells. Digoxin (30 nm) was used as a substrate and verapamil as a control inhibitor. Ethanol, 0.8%, needed for herbal extraction and compatibility with the commercial products, inhibited the net digoxin flux by 18%. E. purpurea influenced to a higher degree the B-A transport of digoxin than the A-B transport. A minor increase in net digoxin flux was observed at low concentrations of E. purpurea, an effect anticipated to be allosteric in nature. At higher concentrations, from 0.4 to 6.36 mg dry weight/mL, a statistically significant linear dose-related decrease was observed in the net digoxin flux, indicating a dose dependent E. purpurea inhibition of P-gp. Both Vmax and Km of the net digoxin flux, calculated to 23.7 nmol/cm2/h and 385 microm, respectively, decreased in the presence of E. purpurea in an uncompetitive fashion. Although the effects of Echinacea purpurea on systemic P-gp mediated drug transport are probably limited, an influence on drug bioavailability can not be excluded.     

The effect of Echinacea purpurea, Astragalus membranaceus and Glycyrrhiza glabra on CD25 expression in humans: a pilot study

This phase 0, double-blind, repeated within subject, randomized pilot study examined CD25 expression on T cells after ingestion of three commonly used herbs: Echinacea purpurea, Astragalus membranaceus and Glycyrrhiza glabra, administered singly and in combination. CD25 expression on T cells was significantly increased for subjects ingesting Echinacea at 24 h with notable increases in activation from Astragalus and Glycyrrhiza. CD25 expression remains elevated with daily use of Echinacea for at least 7 days.     

应用位点特异性引物鉴定白花蛇舌草

目的:建立一种简便实用的白花蛇舌草药材的DNA分子鉴定方法.方法:收集目前市场上出现的白花蛇舌草及伪品共9种,分别提取总DNA,扩增rDNA ITS-2区序列,对比所有样品的该段核苷酸序列,并设计能专一性鉴别白花蛇舌草的位点特异性引物.结果:白花蛇舌草及其混淆品在ITS-2区的序列有显著差异.用位点特异性引物对所有样品DNA进行PCR扩增,只有白花蛇舌草有明显的约392 bp扩增产物,而其它8种植物在同等条件下无阳性产物.结论:所设计的鉴别引物对白花蛇舌草有高度的特异性.     

Naturally occurring insect growth regulators. III. Echinolone, a highly active juvenile hormone mimic from Echinacea angustifolia roots.

A compound, C14H24O2, which induces strong juvenilizing effects in the yellow mealworm, Tenebrio molitor L., following topical application to the pupal stage, was isolated in pure form from the roots of Echinacea angustifolia DC and tentatively identified as dextrorotatory (E)-10-hydroxy-4,10-dimethyl-4,11-dodecadien-2-one. The compound has been named echinolone.     

菊科松果菊属三种药用植物花粉的形态研究

目的：研究３种松果菊属药用植物花粉的形态。方法：对３种花粉进行了光镜（ＬＭ）和扫描电镜（ＳＥＭ）的观察。结果：松果菊属３种花粉形态特征基本一致,但在花粉颜色、大小、萌发沟特征及外壁纹饰上种间存在一定的差异。结论：松果菊属３种植物花粉在形态上存在种间差异。此研究结果为首次报道。     

紫花松果菊亲脂性成分的研究

目的 :分离紫花松果菊的化学成分。方法 :溶剂法和色谱法分离化学成分 ,波谱鉴定其结构。结果 :从地上部分得到 3个化合物 ,1β,6α-二羟基-4(14)桉叶烯(1) ,二十四酸 (4),二十六醇 (5) ;1个混合物 ,(2E ,4E ,8Z ,10E) 异丁基 2 ,4,8,10 十二四烯酰胺 (2 ) ,(2E ,4E ,8Z ,10Z) 异丁基 2 ,4,8,10 十二四烯酰胺 (3)。结论 :化合物1,4和 5为首次从该植物获得。     

Echinacea increases arginase activity and has anti-inflammatory properties in RAW 264.7 macrophage cells,indicative of alternative macrophage activation

Ethnopharmacological relevance

The genus Echinacea is a popular herbal immunomodulator. Recent reports indicate that Echinacea products inhibit nitric oxide (NO) production in activated macrophages.

Aim of the study

In the present study we determined the inhibitory effects of alcohol extracts and individual fractions of alcohol extracts of Echinacea on NO production, and explored the mechanism underlying the pharmacological anti-inflammatory activity.

Materials and methods

Alcohol extracts of three medicinal Echinacea species, Echinacea angustifolia, Echinacea pallida and Echinacea purpurea, were prepared using Soxhlet apparatus and fractionated using HPLC. NO production by LPS activated RAW 264.7 macrophage cells was measured using a Griess reagent and iNOS detected using immunoblotting. In addition, effects on arginase activity were measured in RAW 264.7 cells stimulated with 8-bromo-cAMP +/− LPS.

Results

Alcohol extracts of all three Echinacea species significantly inhibited NO production by lipopolysaccharide (LPS)-activated the RAW 264.7 macrophage cell line; among them Echinacea pallida was the most active. The Echinacea-mediated decrease in NO production was unlikely due to a direct scavenging of NO because the extracts did not directly inhibit NO released from an NO donor, sodium nitroprusside. An immunoblotting assay demonstrated that the extract of Echinacea pallida inhibited inducible nitric oxide synthase (iNOS) protein expression in LPS-treated macrophages. The enzymes iNOS and arginase metabolize a common substrate, l-arginine, but produce distinct biological effects. While iNOS is involved in inflammatory response and host defense, arginase participates actively in anti-inflammatory activation. Arginase activity of RAW 264.7 cells stimulated with 8-bromo-cAMP was significantly increased by alcohol extracts of all three Echinacea species. The polar fraction containing caffeic acid derivatives enhanced arginase activity, while the lipophilic fraction containing alkamides exhibited a potential of inhibiting NO production and iNOS expression.

Conclusions

These results suggest that the anti-inflammatory activity of Echinacea might be due to multiple active metabolites, which work together to switch macrophage activation from classical activation towards alternative activation.