威灵仙类药材HPLC指纹图谱鉴别研究

目的:建立柱果铁线莲的高效液相色谱(HPLC)指纹图谱并与流通的威灵仙药材相比较。方法:基于10批柱果铁线莲,采用RP-HPLC法,以Diamonsil-C18(250mm×4.6mm,5μm)色谱柱为分析柱,流动相为乙腈-0.05%磷酸水,采用梯度洗脱,流速0.8mL/min,210nm波长下对不同来源的药材进行检测。结果:建立了柱果铁线莲药材HPLC-UV指纹图谱共有模式,23个共有峰被标定。并对不同来源的威灵仙类药材的HPLC指纹图谱进行相似度比较,结果具有明显差异。结论:HPLC指纹图谱具有方法简便,重复性好,特征性强等特点,可用于威灵仙类药材的鉴别。     

天胡荽类药材及其混淆品HPLC指纹图谱鉴别研究

目的:建立天胡荽类药材HPLC指纹图谱的鉴别方法。方法:以Diamonsil C18(250 mm×4.6 mm,5μm)色谱柱为分析柱,流动相为乙腈-0.1%磷酸水,采用梯度洗脱,流速1.0 mL/min,340 nm波长下检测。结果:天胡荽类药材的HPLC指纹图谱有明显差异。结论:该方法简便,重复性好,特征性强,可用于天胡荽类药材的鉴别。     

假茼蒿HPLC指纹图谱

目的:建立假茼蒿药材HPLC指纹图谱,对多批次药材进行质量控制。方法:采用Phenomenex C_(18)ODS2色谱柱(4.6 mm×250 mm,5μm),以乙腈-0.1%磷酸水溶液为流动相梯度洗脱,流速1 m L·min~(-1),检测波长328 nm,柱温30℃;建立假茼蒿药材HPLC指纹图谱,对13批药材进行相似度评价,并进行聚类分析。结果:假茼蒿药材HPLC指纹图谱精密度、重复性和稳定性均较好;13批假茼蒿指纹图谱相似度0.97;对13批药材进行聚类分析,总共可聚为4类。结论:将HPLC指纹图谱与聚类分析相结合,可对假茼蒿进行质量评价,可为质量控制提供参考。     

金钱草药材的HPLC指纹图谱研究

目的:建立金钱草药材的HPLC指纹图谱,为金钱草药材的质控提供依据。方法:用水煎-正丁醇萃取-甲醇溶解制备样品溶液,采用RP-HPLC分析,色谱条件为Phenomenex C18色谱柱(4.6 mm×150 mm,5μm),以含0.5%冰醋酸的乙腈(A)-0.5%冰醋酸水(B)梯度洗脱,流速1.0 mL/min,柱温30℃,检测波长255 nm,进样量20μL。利用中药色谱指纹图谱相似度评价系统(2004A版)对不同批次金钱草药材的HPLC图进行比较分析。结果:建立了金钱草药材的HPLC指纹图谱,以芦丁为参照峰,确立了金钱草药材指纹图谱中的13个共有峰。结论:所建立的指纹图谱稳定性和重现性均好,可用于金钱草药材的常规质量控制。     

茜草HPLC-DAD指纹图谱的研究

目的:建立茜草药材HPLC-DAD指纹图谱,提供茜草药材的质量控制方法。方法:采用高效液相色谱法;色谱柱为YMC C18柱(250 mm×4.6 mm,5μm),检测波长280 nm;流动相为乙腈-0.2%磷酸水溶液,梯度洗脱;流速为1.0 mL/min,柱温为25℃。分析时间为60 min。结果:6批茜草药材指纹图谱的平均相似度大于94%。结论:所建立的方法适用于不同产地茜草药材指纹图谱的测定。     

刺五加药材指纹图谱的分析方法研究

目的采用高效液相色谱(HPLC)法建立了刺五加药材指纹图谱。方法色谱柱:Zorbax SB-C18柱(250 mm×4.6mm,5μm),流动相∶乙腈-水(梯度洗脱),检测波长:203 nm,柱温:40℃,流速:1.0 m l/m in,分析时间:90 m in。结果共标示出刺五加药材指纹图谱中24个共有峰。利用指纹图谱相似度测试软件,以夹角余旋为测度,测得10批刺五加药材指纹图谱相似度结果均在0.90~1.00之间。结论本实验所建立的刺五加药材指纹图谱分析方法准确、重复性好,为有效控制刺五加药材的质量提供了依据。     

菝葜药材HPLC指纹图谱的鉴别

目的:建立菝葜药材高效液相色谱(HPLC)指纹图谱鉴别方。方法:采用RP-HPLC,以Diamonsil-C18(4.6 mm×250 mm,5μm)色谱柱为分析柱,流动相乙腈-0.1%磷酸水,梯度洗脱,流速0.6 mL·min-1,330 nm波长下测定菝葜、光叶菝葜的指纹图谱,并作相似度比较分析。结果:建立了菝葜药材HPLC指纹图谱共有模式,18个共有指纹峰被标定,并对菝葜与光叶菝葜药材的HPLC指纹图谱进行相似度比较,结果具有明显差异。结论:HPLC指纹图谱具有方法简便,重复性好,特征性强的特点,可用于菝葜药材的鉴别。     

广西产桑椹HPLC指纹图谱研究

目的:采用HPLC建立广西产桑椹的指纹图谱。方法:采用Agilent Zobax C18(4.6 mm×250 mm,5μm)色谱柱;以乙腈(A)-0.1%磷酸(B)为流动相进行梯度洗脱,检测波长360 nm,流速1.0 mL.min-1,柱温30℃。结果:检测了广西不同来源10批桑椹药材,标定了10个共有峰,相似度较高,建立的桑椹指纹图谱稳定性、重复性好。结论:桑椹HPLC指纹图谱可用于桑椹药材的鉴别及质量控制。     

栀子HPLC数字化指纹图谱相对定量方法研究

目的:建立栀子HPLC数字化指纹图谱并以对照药材为参照对相对定量方法进行研究.方法:采用反相高效液相色谱法,使用CenturySIL C18 BDS柱(20 cm x4.6mm,5 μm),以1%醋酸水-1%醋酸乙腈为流动相,低压线性梯度洗脱,检测波长265nm,柱温(30.0±0.15)℃,测定10批不同产地栀子HPLC指纹图谱和栀子对照药材指纹图谱.以栀子对照药材供试液不同进样量时各指纹峰的峰面积分别对表观进样量(mg)进行线性回归,用"中药色谱指纹图谱超信息特征数字化评价系统"软件计算得对照药材折合标准曲线.结果:以栀子苷峰为参照物峰,确定35个共有峰,建立了栀子HPLC数字化指纹图谱.以栀子对照药材为参照计算出各产地栀子相当于对照药材的折合百分含量以及所有指纹成分相当于对照药材中相应成分的折合百分含量.结论:所建立的指纹图谱相对定量方法可衡量不同产地栀子药材中指纹成分的含量,可作为栀子药材质量控制的依据.该方法对深入研究单一指纹峰的相对含量具有较重要意义,也为丰富中药色谱指纹图谱的定量评价方法提供了重要参考价值.     

刺五加HPLC数字化指纹图谱研究

目的:建立刺五加HPLC数字化指纹图谱。方法:采用反相高效液相色谱法,使用Century SIL BDS(20 cm×4.6mm,5μm)柱,以1%醋酸水-1%醋酸乙腈为流动相,线性梯度洗脱,流速1.0 mL/mL,检测波长265 nm,柱温(30.0±0.15)℃,进样量10μL。以定性、定量相似度和色谱指纹图谱指数等数字化指标评价药材质量。结果:以绿原酸峰为参照物峰,确定31个共有指纹峰,建立了刺五加HPLC数字化指纹图谱。应用色谱指纹图谱指数F和相对指数Fr参数对不同产地刺五加HPLC指纹图谱的超信息特征进行了数字化评价。结论:所建立的HPLC指纹图谱具有较好的精密度和重现性,适用于刺五加药材的质量控制。     

Effect of feeding Mucuna pruriens on helminth parasite infestation in lambs

Ethnopharmacological relevance

Mucuna pruriens is a tropical legume anecdotally reputed to have anthelmintic properties. This study was conducted to examine the validity of such claims.

Aim of the study

The aim of this study was to determine if ingestion of Mucuna seeds reduces helminth parasite infestation in lambs.

Materials and methods

Thirty-six Dorper × Katahdin ram lambs were assigned to three treatments, a cottonseed meal based control diet, a diet in which Mucuna replaced cottonseed meal and the control diet with levamisole (7.5 mg/kg body weight) administration. All diets were isonitrogenous and isocaloric. The 12 lambs in each treatment were assigned randomly to 4 pens, each containing 3 lambs. Lambs were trickle infected three times per week by gavage with infectious Haemonchus contortus larvae (2000 larvae/lamb) for 3 weeks.

Results

Levamisole treatment decreased fecal egg counts by 87% and abomasal worm counts by 83%. Mucuna intake did not statistically affect fecal egg counts or abomasal worm counts, though numerical (P > 0.10) reductions of 7.4% and 18.1%, respectively were evident. Anemia indicators, feed intake, and lamb growth were unaffected by treatment.

Conclusions

Levamisole reduced the Haemonchus parasite burden in lambs significantly but feeding Mucuna reduced the burden by levels unlikely to eliminate the clinical effects of parasitism.     

Investigation of plant extracts in traditional medicine of the Brazilian Cerrado against protozoans and yeasts

Aim of the study

To investigate the activities of the 217 plant extracts in traditional medicine of the Brazilian Cerrado against protozoans and yeasts.

Materials and methods

Plant extracts were prepared by the method of maceration using solvents of different polarities. The growth inhibition of chloroquine-resistant Plasmodium falciparum strain (FcB1) was determined by measuring the radioactivity of the tritiated hypoxanthine incorporated. Activity against Leishmania (Leishmania) chagasi and Trypanosoma cruzi was measured by the MTT colorimetric assay. The antifungal tests were carried out by using the CLSI method. The active extracts were tested also by cytotoxicity assay using NIH-3T3 cells of mammalian fibroblasts.

Results

Two hundred and seventeen extracts of plants were tested against Plasmodium falciparum. The eleven active extracts, belonging to eight plant species were evaluated against L. (L.) chagasi, Trypanosoma cruzi, yeasts and in NIH-3T3 cells. The results found in these biological models are consistent with the ethnopharmacological data of these plants. The ethyl acetate extract of Diospyros hispida root showed IC₅₀ values of 1 μg/mL against Plasmodium falciparum. This extract demonstrated no toxicity against mammalian cells, resulting in a significant selectivity index (SI) of 435.8. The dichloromethane extract of Calophyllum brasiliense root wood was active against Cryptococcus gattii LMGO 01 with MIC of 1.95 μg/mL; and Candida albicans ATCC 10231 and Candida krusei LMGO 174, both with MIC of 7.81 μg/mL. The same extract was also active against Plasmodium falciparum and L. (L.) chagasi with IC₅₀ of 6.7 and 27.6 μg/mL respectively. The ethyl acetate extract of Spiranthera odoratissima leaves was active against Cryptococcus gattii LMGO 01 with MIC of 31.25 μg/mL, and against Plasmodium falciparum with IC₅₀ of 9.2 μg/mL and Trypanosoma cruzi with IC₅₀ of 56.3 μg/mL.

Conclusion

The active extracts for protozoans and human pathogenic yeasts are considered promising to continue the search for the identification and development of leading compounds.     

真菌的中药干预研究近5年进展

近年来真菌感染率逐年上升,传统抗真菌药物易产生耐药性,而中药在防治真菌感染方面具有一定的优势。本文就近5年来中药对白念珠菌、皮肤癣菌、曲霉菌、马拉色菌、串珠镰孢菌、申克孢子丝菌、新生隐球菌及真菌生物膜的干预研究进展进行综述。     

α-Glucosidase inhibitory activity of selected Philippine plants

Ethnopharmacological relevance

Antidesma bunius Spreng. (Phyllantaceae), Averrhoa bilimbi L. (Oxalidaceae), Biophytum sensitivum (L.) DC. (Oxalidaceae), Ceriops tagal (Perr.) C.B. Rob. (Rhizophoraceae), Kyllinga monocephala Rottb. (Cyperaceae), and Rhizophora mucronata Lam. (Rhizophoraceae) are used as remedies to control diabetes. In the present study, these plants were screened for their potential α-glucosidase inhibitory activity.

Materials and methods

The 80% aqueous ethanolic extracts were screened for their α-glucosidase enzyme inhibitory activity using yeast alpha glucosidase enzyme.

Results

Except for A. bilimbi with IC₅₀ at 519.86±3.07, all manifested a significant enzyme inhibitory activity. R. mucronata manifested the highest activity with IC₅₀ at 0.08±1.82 μg mL⁻¹, followed by C. tagal with IC₅₀ at 0.85±1.46 μg mL⁻¹ and B. sensitivum with IC₅₀ at 2.24±1.58 μg mL⁻¹.

Conclusion

This is the first report on the α-glucosidase inhibitory effect of the six Philippine plants; thus, partly defining the mechanism on why these medicinal plants possess antidiabetic properties.     

European medicinal polypores – A modern view on traditional uses

Ethnopharmacological relevance

In particular five polypore species, i.e. Laetiporus sulphureus, Fomes fomentarius, Fomitopsis pinicola, Piptoporus betulinus, and Laricifomes officinalis, have been widely used in central European folk medicines for the treatment of various diseases, e.g. dysmenorrhoea, haemorrhoids, bladder disorders, pyretic diseases, treatment of coughs, cancer, and rheumatism. Prehistoric artefacts going back to over 5000 years underline the long tradition of using polypores for various applications ranging from food or tinder material to medicinal–spiritual uses as witnessed by two polypore species found among items of Ötzi, the Iceman. The present paper reviews the traditional uses, phytochemistry, and biological activity of the five mentioned polypores.

Materials and methods

All available information on the selected polypore taxa used in traditional folk medicine was collected through evaluation of literature in libraries and searches in online databases using SciFinder and Web of Knowledge.

Results

Mycochemical studies report the presence of many primary (e.g. polysaccharides) and secondary metabolites (e.g. triterpenes). Crude extracts and isolated compounds show a wide spectrum of biological properties, such as anti-inflammatory, cytotoxic, and antimicrobial activities.

Conclusions

The investigated polypores possess a longstanding ethnomycological tradition in Europe. Here, we compile biological results which highlight their therapeutic value. Moreover, this work provides a solid base for further investigations on a molecular level, both compound- and target-wise.     

厚朴与凹叶厚朴群体遗传学研究

目的:对厚朴与凹叶厚朴的群体遗传学进行研究,为中药厚朴的质量控制提供分子生药学方面的依据。方法:对厚朴与凹叶厚朴15个居群应用2个叶绿体基因间序列psbA-trnH和trnL-trnF进行PCR扩增并测序,计算厚朴与凹叶厚朴单倍型频率,用程序HaploNst分析遗传多样性和遗传结构,应用TCS version 1.13软件构建单倍型网状进化树。结果:厚朴与凹叶厚朴均无特有单倍型存在,但单倍型频率存在显著差异,已开始出现遗传分化的趋势,NST略大于GST。结论:厚朴与凹叶厚朴在遗传上已出现遗传分化的趋势,但尚未完全分化成彼此独立的单系。     

Antiplasmodial activity of extracts from seven medicinal plants used in malaria treatment in Cameroon

Aim of the study

In a search for new plant-derived biologically active compounds against malaria parasites, we have carried out an ethnopharmacological study to evaluate the susceptibility of cultured Plasmodium falciparum to extracts and fractions from seven Cameroonian medicinal plants used in malaria treatment. We have also explored the inhibition of the Plasmodium falciparum cysteine protease Falcipain-2.

Materials and methods

Plant materials were extracted by maceration in organic solvents, and subsequently partitioned or fractionated to afford test fractions. The susceptibility of erythrocytes and the W2 strain of Plasmodium falciparum to plant extracts was evaluated in culture. In addition, the ability of annonaceous extracts to inhibit recombinant cysteine protease Falcipain-2 was also assessed.

Results and discussion

The extracts showed no toxicity against erythrocytes. The majority of plant extracts were highly active against Plasmodium falciparumin vitro, with IC₅₀ values lower than 5 μg/ml. Annonaceous extracts (acetogenin-rich fractions and interface precipitates) exhibited the highest potency. Some of these extracts exhibited modest inhibition of Falcipain-2.

Conclusion

These results support continued investigation of components of traditional medicines as potential new antimalarial agents.     

中国石斛属植物文献计量研究

石斛是珍稀濒危中药材,目前正处于快速发展阶段。为全面了解我国石斛属植物研究的历史和发展现状,作者以1954～2010年"中国知网中国学术期刊网络出版总库"收录的石斛研究文献为依据,采用文献计量学的原理和方法,对我国石斛属植物研究文献从文献年代分布、期刊分布与被引频率、主题分布、研究对象分布、作者与研究机构分布等方面进行了统计与分析。结果表明,我国石斛研究明显分为起步(2个)、停滞、平稳发展、快速上升5个阶段;期刊分布存在离散性与集中性并存的现象,已形成核心期刊研究群,并以《中国中药杂志》、《中草药》和《陕西中医》为代表;研究主题广泛涉及临床与药理、组织培养与种苗繁育、成分分析等多个领域,已经形成比较稳定的研究机构和团队,但研究对象差异显著,以铁皮石斛、金钗石斛和霍山石斛最为集中。我国石斛属植物的研究已取得显著成果,但种植产业发展缓慢,供需矛盾突出,预计种苗繁育与人工种植、产品开发、化学与药理等方面是未来的研究热点,其文献报道仍将进一步上升。     

列当属、肉苁蓉属和草苁蓉属植物传统药物学调查

通过野外资源调查、整理各大标本馆标本原始记录和查阅文献记载等方法,系统整理、总结、归纳了列当属、肉苁蓉属和草苁蓉属民族药用植物种类、功效及民间使用情况,结果表明列当属6种药用植物在4个少数民族间作为7种民族药应用,草苁蓉属2种药用植物在8个少数民族间作为10种民族药应用,肉苁蓉属2种药用植物在3个少数民族间作为3种民族药应用,且同种药用植物常作不同民族药;发现3属植物的传统疗效主要集中在补肾壮阳、止血和止痛3个方面,并且该传统疗效与现代药理研究结果基本吻合。因此深入研究植物种类丰富的列当属植物资源对缓解肉苁蓉植物资源匮乏局面和扩大药源具有积极意义。     

金银花类药用植物IspE和IspH基因克隆和生物信息学分析

目的:克隆金银花类药用植物4-二磷酸胞苷-2-C-甲基赤藓糖激酶(4-diphosphocytidyl-2-C-methyl-D-erythritol kinase,IspE)和4-羟基-3-甲基-2-邻苯基二磷酸还原酶(4-hydroxy-3-methylbut-2-enyl diphosphate reductase,IspH)基因,并对其基因序列、蛋白特性和转录活性进行分析、比较.方法:从忍冬Lonicera japonica转录组测序结果中分析获得了IspE,IspH基因.分别以忍冬、红白忍冬L.japonica var.chinensis、红腺忍冬L.hypoglauca和水忍冬L.dasystyla新鲜花蕾为材料,利用RT-PCR技术克隆获得了4种金银花类药用植物IspE和IspH基因的全长cDNA.运用生物信息学分析软件,预测编码蛋白的结构和功能,并通过RT-PCR检测IspE和IspH在忍冬、红白忍冬、红腺忍冬、水忍冬花蕾中的转录情况.结果:金银花类药用植物IspE基因含有1个完整的开放阅读框,长度为1 221 bp,编码406个氨基酸;IspH含有一个完整的开放阅读框,长度为1 380 bp,编码459个氨基酸.IspE和IspH均为非分泌蛋白,均定位于叶绿体中.RT-PCR分析结果表明在忍冬、红腺忍冬和水忍冬的花蕾中IspE,IspH基因的转录水平没有显著差异,但红白忍冬花蕾中IspE,IspH基因的转录水平均显著高于忍冬.结论:克隆获得忍冬、红白忍冬、红腺忍冬和水忍冬中IspE,IspH基因,并证实了其在不同金银花类药用植物中的表达,为进一步研究IspE,IspH基因对萜类化合物生物合成和花香气以及颜色的影响奠定了基础.