药用真菌活性次级代谢产物研究

药用真菌作为我国传统中医药体系的重要组成部分,有着悠久的历史。它们能够产生丰富的活性次级代谢产物,具有神经保护、抗肿瘤、降血脂等诸多药效。然而,目前对大部分药用真菌次级代谢产物的化学和生物学研究较少。本课题组主要以抗肿瘤、抗炎、抗菌以及抗病毒等生物活性为指导,选择重要药用真菌进行化学研究,构建我国特色药用真菌代谢产物库。文章主要介绍近4年我们在这方面的研究进展。     

中国药用植物资源开发利用研究的回顾与展望

本文对药用植物资源开发的方法学进行了讨论,并简要综述了近年来我国在药用植物资源开发利用研究方面的成就,还就药用植物资源开发利用中的一些重要问题:生物多样性的保护;资源数据库的建立及管理;新技术和新方法的引进以及中药外向过程中资源的宏观调控管理等进行了讨论。     

药用植物内生真菌研究现状和发展趋势

药用植物内生真菌普遍存在于健康植物组织和器官中,种类繁多,分布广泛。到目前为止,人们已从大量药用植物中分离出不同类型的内生真菌,这些植物广泛分布于除南极洲以外的各种陆地生态系统中。随着现代科学技术的迅速发展,药用植物内生真菌的研究也取得了长足的进步。由于内生真菌对于药用植物的重要性,其相关研究也受到了世界各国学者的高度关注。本文总结现阶段药用植物内生真菌相关研究,并对未来研究的发展趋势做出展望。     

Endophytic fungi with anti-microbial,anti-cancer and anti-malarial activities isolated from Thai medicinal plants

A total of 81 Thai medicinal plant species collected from forests in four geographical regions of Thailand were examined for the presence of endophytic fungi with biological activity. Of 582 pure isolates obtained, 360 morphologically distinct fungi were selected for cultivation on malt Czapek broth and yeast extract sucrose broth, from which extracts were tested for biological activity. Extracts of 92 isolates could inhibit Mycobacterium tuberculosis (MIC 0.0625–200 μg ml⁻¹) when tested by the microplate Alamar blue assay, while extracts of six inhibited Plasmodium falciparum (IC₅₀ of 1.2–9.1 μg ml⁻¹) as determined by the [³H]hypoxanthine incorporation method. Strong anti-viral activity against Herpes simplex virus type 1 was observed in 40 isolates (IC₅₀ of 0.28–50 μg ml⁻¹). The sulphorhodamine B assay for activity against cancer cell lines revealed that 60 were active against human oral epidermoid carcinoma cells (EC₅₀ 0.42–20 μg ml⁻¹) and 48 against breast cancer cells (EC₅₀ 0.18–20 μg ml⁻¹). Bioactivity profile was affected by the type of culture medium. Given the high incidence of bioactive extracts and the fact that most of the isolated fungi could not be identified due to lack of spore formation, the results suggested that Thai medicinal plants can provide a wide variety of endophytes that might be a potential source of novel bioactive compounds. This revised version was published online in August 2006 with corrections to the Cover Date.     

Database on pharmacophore analysis of active principles, from medicinal plants

Plants continue to be a major source of medicines, as they have been throughout human history. In the present days, drug discovery from plants involves a multidisciplinary approach combining ethnobotanical, phytochemical and biological techniques to provide us new chemical compounds (lead molecules) for the development of drugs against various pharmacological targets, including cancer, diabetes and its secondary complications. In view of this need in current drug discovery from medicinal plants, here we describe another web database containing the information of pharmacophore analysis of active principles possessing antidiabetic, antimicrobial, anticancerous and antioxidant properties from medicinal plants. The database provides the botanical, taxonomic classification, biochemical as well as pharmacological properties of medicinal plants. Data on antidiabetic, antimicrobial, anti oxidative, anti tumor and anti inflammatory compounds, and their physicochemical properties, SMILES Notation, Lipinski's properties are included in our database. One of the proposed features in the database is the predicted ADMET values and the interaction of bioactive compounds to the target protein. The database alphabetically lists the compound name and also provides tabs separating for anti microbial, antitumor, antidiabetic, and antioxidative compounds. AVAILABILITY: http://www.hccbif.info /     

Phenolic Profiling,Antioxidant, and Antibacterial Activities of Selected Medicinal Plants from Tunisia

Phytochemical screening of aqueous extract from six medicinal wild plants grown in South-eastern of Tunisia: Atriplex halimus, Teucrium polium, Moricandia arvensis, Deverra tortuoa, Haplophyllum tuberculatum and Polygonum equisetiforme were evaluated. Both decoction and ultrasound assisted extraction were used. Antioxidant, antibacterial proprieties, and phenolic profiling, using LC-ESI-MS method, were assessed. Total polyphenols, flavonoids, and condensed tannins contents ranged from 7.47±0.19 to 22.25±0.49 mg GAE/g Dw, 5.47±0.06 to 7.55±0.07 mg RE/g Dw, and 0.33±0.02 to 19.43±0.64 mg TAE/g Dw, respectively. Moreover, the reducing power and DPPH tests showed that P. equisetiforme (EC₅₀: 12.50±0.50 μg/ml; DPPH⋅⁺: 213.49±4.24 mg TEAC/g DW), T. polium (EC₅₀: 25.00±1.00 μg/ml; DPPH⋅⁺: 181.39±9.47 mg TEAC/g DW) as well as H. tuberculatum (EC₅₀: 56.25±0.25 μg/ml; DPPH⋅⁺: 177.83±5.85 mg TEAC/g DW) extracts were the most effective natural antioxidants. For anti-bacterial activity, the ultrasonic extract of H. tuberculatum showed the highest activity against both P. aeruginosa (13.50±0.71 mm) and S. aureus (13.00±0.00 mm) at 10 mg/ml. Furthermore 24 phenolic compounds were identified, with predominance of quinic acid, gallic acid, protocatechuic acid, syringic acid, p-coumaric acid, trans-ferulic acid, catechin (+), trans-cinnamic and silymarin. These results were further consolidated by to heatmap clustering with P. equisetiforme, H. tuberculatum, T. polium as the main antioxidant and antibacterial sources which supports their domestication and industrial use.     

Antifungal activity of six Saudi medicinal plant extracts against five phyopathogenic fungi

Six medicinal plants such as Amaranthus spinosus, Barbeya oleoides, Clutia lanceolata, Lavandula pubescens, Maerua oblongifolia and Withania somnifera collected from different locations in the southwestern part of Saudi Arabia were tested for antifungal activities against five plant pathogenic fungi causing serious diseases of vegetable crops. These fungi were Alternaria brassicae, Alternaria solani, Botrytis fabae, Fusarium solani and Phytophthora infestans. Aqueous plant extracts reduced mycelial growth and inhibited spore germination of all fungi tested. It is clear that the aqueous extract of Lavandula pubescens leaves was the best for controlling all phytopathogenic fungi under study. These results suggested that medicinal plant extracts play an important role in controlling the phytopathogenic fungi.     

Towards the scientific validation of traditional medicinal plants

A large proportion of the population of developing countries usestraditional medicine alone, or in combination with Western drugs to treat awide variety of ailments. There has seldom been effective collaborationbetween the traditional and Western medical practitioners, largely due tothe perception that the use of traditional and herbal medicines has noscientific basis. With the renewed interest from Western countries in herbalremedies, and the increasingly urgent need to develop new effective drugs,traditionally used medicinal plants have recently received the attention ofthe pharmaceutical and scientific communities. This involves the isolationand identification of the secondary metabolites produced by the plants andused as the active principles in medical preparations. Research into thescientific validation of southern African medicinal plants used in thetreatment of pain and inflammation, hypertension and parasitic diseasesincluding those with anthelmintic, anti-amoebic, anti-bacterial and anti-bilharzia activity, is the current focus of studies conducted at the ResearchCentre for Plant Growth and Development.     

三种水生植物内生真菌多样性及其抗真菌活性

从华凤仙(Impatiens chinensis L.)、问荆(Equisetum arvense L.)和轮叶狐尾藻(Myriophyllum verticillatum L.) 3种植物中共分离到内生真菌155株。经鉴定, 它们分属于26个不同的分类单元。以杨桃炭疽菌(Colletotrichum gloeosporioides)、香蕉疫霉(Phytophthora nicotianae)和腐皮镰刀菌(Fusarium solani)等6种植物病原真菌为指示菌, 对3种水生植物内生真菌代谢产物进行抗菌活性研究。结果表明, 有37株内生真菌(占分离菌株总数的23.9%)显示出对一种或多种病原菌的抑菌活性。来自华凤仙、问荆和轮叶狐尾藻的抗菌活性菌株比例分别为29.2%、13.9%及37.1%, 都远低于已报道的陆生植物内生真菌中活性菌株的比例。抗菌活性菌株主要分布于枝孢属(Cladosporium)、木霉属(Trichoderma)和地霉属(Geotrichum)等5个属中。     

The Pacific Islands: a biotechnology resource bank of medicinal plants and traditional intellectual property

Traditional medicines, the mainstay of medical treatment for virtually all minor ailments in many developing countries, have been tapped for the production of new therapeutics in the sustenance of human health and well-being. Vulnerable to the vicissitudes of globalization issues such as intellectual property rights, trade and gender are of relevance in the Pacific region that is a source of some unique traditional healing systems.     

Screening for Endophytic Fungi with Antitumour and Antifungal Activities from Chinese Medicinal Plants

Summary One hundred and thirty endophytic fungi isolated from 12 Chinese traditional medicinal plants collected at Yuanmou county and Dawei Mountain, Yunnan province, southwest China, were tested for antitumour and antifungal activities by MTT assay on human gastric tumour cell line BGC-823 and the growth inhibition test against 7 phytopathogenic fungi. The results showed that fermentation broths from 9.2% of the isolates exhibited antitumour activity and 30% exhibited antifungal activity, moreover, some of them exhibited broad-spectrum antifungal activity. The active isolates were identified to 32 taxa. The results indicate that the endophytic fungi of Chinese traditional medicinal plants are promising sources of novel bioactive compounds.     

Endophytic fungi from <Emphasis Type="Italic">Nerium oleander</Emphasis> L (Apocynaceae): main constituents and antioxidant activity

Diverse endophytic fungi exist within plant aerial tissues, with a global estimate of up to a million undescribed species. These endophytes constitute a rich bio-resource for exploration to discover new natural products. Here we investigate fungal endophytes associated with a medicinal plant, Nerium oleander L. (Apocynaceae). A total of 42 endophytic fungal strains were isolated from the host plant. Total antioxidant capacity, xanthine oxidase inhibitory activity, antimicrobial activity, and total phenolic content (TPC) were evaluated for 16 representative fungal cultures grown in improved Czapek’s broth and for the host plant. The total antioxidant capacities and phenolic contents of the fungal cultures ranged from 9.59 to 150.79 μmol trolox/100 mL culture, and from 0.52 to 13.95 mg gallic acid/100 mL culture, respectively. The fungal culture of an endophytic strain Chaetomium sp. showed the strongest antioxidant capacity, contained the highest level of phenolics, and to some extent inhibited xanthine oxidase activity with an IC₅₀ value of 109.8 μg/mL. A significant positive correlation was found between antioxidant capacity and TPC in the tested samples. Most of the endophytic fungal cultures tested have a wide range of antimicrobial activities, which were not very strong, but much better than those of the host plant. The major bioactive constituents of the fungal cultures were investigated using LC-ESI-MS and GC-MS, and preliminary identification detected phenolics (e.g. phenolic acids and their derivatives, flavonoids) and volatile and aliphatic compounds. This study shows that the endophytic fungi isolated from N. oleander can be a potential antioxidant resource.     

Molecular characterization and antimicrobial activity of endophytic fungi from coffee plants

Endophytic filamentous fungi from coffee plants (Coffea arabica and C. robusta) deposited in the Brazilian Collection of Environmental and Industrial Microorganisms (CBMAI) were characterized taxonomically by using molecular tools and investigated concerning their antimicrobial activity against different human pathogenic bacteria. Thirty-seven out of 39 CBMAI strains investigated were identified to at least at genus level by ITS and rDNA D1/D2 sequencing and phylogenetic analyses. Bioactivity screening of fungal extracts against Salmonella choleraesuis (CBMAI 484), Staphylococcus aureus (CBMAI 485), Pseudomonas aeruginosa (CBMAI 489) and against four different Escherichia coli serotypes showed that 17 fungi inhibited at least one of the bacteria studied. The endophytic fungi Trichoderma harzianum (CBMAI 43), Guignardia sp. (CBMAI 69) and Phomopsis sp. (CBMAI 164) inhibited from four to five bacterial species, while five fungi were active against all pathogenic bacteria tested and were identified as Aspergillus versicolor (CBMAI 46), Fusarium oxysporum (CBMAI 53), Glomerella sp. (CBMAI 63) and Cladosporium spp. (CBMAI 64 and CBMAI 66). The Minimal Inhibitory Concentration (MIC) for the fungus extracts varied from 0.025 to 1.0 mg ml⁻¹, demonstrating antimicrobial potential of some of these fungi.     

Molecular profiling of marine endophytic fungi from green algae: Assessment of antibacterial and anticancer activities

Bioactive natural metabolites, especially from the marine endophytic fungi, are largely unexplored. Endophytic fungi are being increasingly recognized as a group of organisms that produce novel metabolites of industrial importance. This study investigated the anticancer and antibacterial potential of the marine algal endophyte, Penicillium chrysogenum. The different organic solvent extracts of the endophytic fungi grown on different growth medium were analyzed for anticancer and antibacterial activities. The highest inhibitory activity was observed for the ethyl acetate (EA) extract of the culture filtrate grown in potato dextrose broth (PDB) for 21 days, against the tested human breast cancer cell (MCF-7) line. Similarly, the PDB-EA extract showed an appreciable activity against the human pathogens. The biochemical analysis of the Cha EA metabolites revealed terpenoids, steroids, phenolics and flavones. Gas Chromatography (GCMS) data revealed several bioactive compounds such as anthraquinone and cinnamic acid. The Cha EA extract induced membrane damage and thus, apoptosis in MCF-7cells. The secondary metabolites produced by these marine endophytic fungi have contributed to considerable anticancer and antimicrobial activities and hence, this study is an evidence of potential sources of antimicrobial and anticancer compounds from Penicillium chrysogenum.     

药用昆虫蜣螂对灵芝发酵产物体外抗肿瘤活性的影响

采用体外高通量筛选技术检测了补加药用昆虫蜣螂前后灵芝发酵产物的体外抗肿瘤活性。结果表明,补加和不补加蜣螂发酵后所得的胞内和胞外三萜样品对肉瘤细胞L290、肠癌细胞SW620、血癌细胞K562和肝癌细胞BEL7402都有显著的抑制作用(P<0.05)。在补加蜣螂发酵后,灵芝胞内三萜的抑制作用没有得到增强;但胞外三萜样品对BEL7402细胞的抑制作用得到了增强,补加和不补加蜣螂发酵后所得胞外三萜的抑制率分别为41.74%和32.37%(P<0.05)。由于补加蜣螂发酵后,新生成了胞外三萜lucidone C,因而对lucidone C的抗BEL7402肝癌活性进行了试验。结果显示,lucidone C在100μg/mL时,对BEL7402的抑制率为50.37%,提示lucidone C可能增强了补加蜣螂后灵芝胞外总三萜对BEL7402细胞的抑制作用。     

Phenolic composition and antioxidant properties of some traditionally used medicinal plants affected by the extraction time and hydrolysis

Introduction – Polyphenolic phytochemicals in traditionally used medicinal plants act as powerful antioxidants, which aroused an increasing interest in their application in functional food development. Objective – The effect of extraction time (5 and 15 min) and hydrolysis on the qualitative and quantitative content of phenolic compounds and antioxidant capacity of six traditionally used medicinal plants (Melissa officinalis L., Thymus serpyllum L., Lavandula officinalis Miller, Rubus fruticosus L., Urtica dioica L., and Olea europea L.) were investigated. Methodology – The content of total phenols, flavonoids, flavan‐3‐ols and tannins was determined using UV/Vis spectrophotometric methods, while individual phenolic acids, flavones and flavonols were separated and detected using HPLC analysis. Also, to obtain relevant data on the antioxidant capacity, two different assays, (2,2‐azino‐bis(3‐ethylbenzthiazoline‐6‐sulphonic acid) (ABTS) radical scavenging and ferric reducing/antioxidant power (FRAP) assays were used. Results – The extraction efficiency of phenolics, as well as the antioxidant capacity of plant extracts, was affected by both prolonged extraction and hydrolysis. The overall highest content of phenolic compounds was determined in hydrolyzed extract of blackberry leaves (2160 mg GAE/L), followed by the non‐hydrolyzed extract of lemon balm obtained after 15 min of extraction (929.33 mg GAE/L). The above extracts also exhibited the highest antioxidant capacity, while extracts of olive leaves were characterized with the lowest content of phenolic compounds, as well as the lowest antioxidant capacity. The highest content of rosmarinic acid, as the most abundant phenolic compound, was determined in non‐hydrolyzed extract of lemon balm, obtained after 15 min of extraction. Although the hydrolysis provided the highest content of polyphenolic compounds, longer extraction time (15 min) was more efficient to extract these bioactives than shorter extraction duration (5 min). Conclusion – The distribution of detected phenolic compounds showed a wide variability with regard to their botanical origin. Examined medicinal plants showed to be a valuable supplement to a daily intake of bioactive compounds. Copyright © 2010 John Wiley & Sons, Ltd.     

Screening of anti-Helicobacter pylori herbs deriving from Taiwanese folk medicinal plants

In this study, extracts from 50 Taiwanese folk medicinal plants were examined and screened for anti-Helicobacter pylori activity. Ninety-five percent ethanol was used for herbal extraction. Paederia scandens (Lour.) Merr. (PSM), Plumbago zeylanica L. (PZL), Anisomeles indica (L.) O. Kuntze (AIOK), Bombax malabaricum DC. (BMDC) and Alpinia speciosa (J. C. Wendl.) K. Schum. (ASKS) and Bombax malabaricum DC. (BMDC) all demonstrated strong anti-H. pylori activities. The minimum inhibitory concentration values of the anti-H. pylori activity given by the five ethanol herb extracts ranged from 0.64 to 10.24 mg ml(-1). Twenty-six herbs, including Artemisia argvi Levl. et Vant (AALEV), Phyla nodiflora (Linn.) Greene (PNG) and others, showed moderate anti-H. pylori activity. The additional 19 herbs, including Areca catechu Linn. (ACL), Euphorbia hirta Linn. (EHL) and Gnaphalium adnatum Wall. ex DC. (GAWEDC), possessed lower anti-H. pylori effects. About half of the Taiwanese folk medicinal plants tested, demonstrated to possess higher anti-H. pylori activity.     

四种植物内生真菌的分离及其抗肿瘤活性的筛选

从银杏、合欢、臭椿和苦楝的树皮中分离到61株内生真菌。MTT法检测显示在45.9%的菌株发酵液粗提物在200 μg/mL时对人食道癌细胞EC109的生长抑制率大于50%。其中6株内生真菌(YX5, YX17, YX36, KL1, CC1, CC5)在50 μg/mL时仍具有较高的细胞毒活性, 且活性成分主要分布在菌株的发酵液中。合欢内生真菌中的活性菌株比例最小, 在50 μg/mL时均未显示任何对EC109细胞的抑制作用。银杏中的高活性菌株比例最大, 占银杏内生真菌的15.8%。银杏分离菌株YX5的活性最为突出, 其发酵液粗提物对3种肿瘤细胞EC109, HONE1和HeLa的抑制率IC50分别是18.3 μg/mL、3.6 μg/mL和6.5 μg/mL。研究结果表明, 作为抗肿瘤药物的潜在来源银杏内生真菌值得关注。     

Structure-radical scavenging activity relationships of phenolic compounds from traditional Chinese medicinal plants

Traditional Chinese medicinal plants associated with anticancer contain a wide variety of natural phenolic compounds with various structural features and possessing widely differing antioxidant activity. The structure-radical scavenging activity relationships of a large number of representative phenolic compounds (e.g., flavanols, flavonols, chalcones, flavones, flavanones, isoflavones, tannins, stilbenes, curcuminoids, phenolic acids, coumarins, lignans, and quinones) identified in the traditional Chinese medicinal plants were evaluated using the improved ABTS*+ and DPPH methods. Different categories of tested phenolics showed significant mean differences in radical scavenging activity. Tannins demonstrated the strongest activity, while most quinones, isoflavones, and lignans tested showed the weakest activity. This study confirmed that the number and position of hydroxyl groups and the related glycosylation and other substitutions largely determined radical scavenging activity of the tested phenolic compounds. The differences in radical scavenging activity were attributed to structural differences in hydroxylation, glycosylation and methoxylation. The ortho-dihydroxy groups were the most important structural feature of high activity for all tested phenolic compounds. Other structural features played a modified role in enhancing or reducing the activity. Within each class of phenolic compounds, the structure-activity relationship was elucidated and discussed. This study reveals the structure-activity relationships of a large series of representative natural phenolic compounds more systematically and fully than previous work. Structure-radical scavenging activity relationships of some natural phenolics identified in the medicinal plants were evaluated for the first time.     

药用植物内生真菌的多样性及生物功能研究进展

药用植物内生真菌资源丰富,其代谢产物常具有抗肿瘤、抗氧化、抑菌等作用,能产生药用植物生长调节物质及与宿主相同或类似的次生代谢产物,从而成为近年来的研究热点。本文对药用植物内生真菌的分离鉴定、多样性、生物活性及生物学功能等方面进行综述,以期为今后筛选及利用有效的药用植物内生真菌奠定基础。