RESUMO
Two new cyclopentapeptides, basidiosins A and B (1 and 2) were isolated from the mycelia extracts of entomophthoralean fungus Basidiobolus meristosporus RCEF 4516. The structures were determined based on spectroscopic methods, and the absolute config urations were assigned by Marfey's method on their acid hydrolyzates. Compounds 1 and 2 were identified as cyclo(L-Thr-L-Leu- L-Ile-D-Tyr-D-Thr) and cyclo(L-Thr-L-Leu-L-Val-D-Val-D-Ser), respectively. They were evaluated for the biological activities including antibacterial, antifungal and antioxidative activities. Furthermore, the biosynthetic pathway of 1 was proposed by bioinformatic analysis. This is the first study on the isolation of natural products from Basidiobolus fungus.
Assuntos
Produtos Biológicos/farmacologia , Entomophthorales/química , Antifúngicos/isolamento & purificação , Antifúngicos/farmacologia , Produtos Biológicos/isolamento & purificação , China , Entomophthorales/genética , Florestas , Estrutura Molecular , Família Multigênica , Micélio/química , Fragmentos de Peptídeos/isolamento & purificação , Fragmentos de Peptídeos/farmacologia , Microbiologia do SoloRESUMO
Isaria cicadae is one of the fungi used in traditional Chinese medicine with the longest tradition. It is used not only as a herbal medicine but also as a health food in Asia, together with cultured cordyceps and mycelia of the fungus used as substitute. However, the differences in their metabolite are unknown. Using a high-performance liquid chromatography-mass spectrometry (HPLC-MS)-based metabolomic method, we found that the fungus varies in its metabolism during growth on wild insects, artificially raised insects and artificial medium. There were 109 discriminatory metabolites detected in the samples by orthogonal projection to latent structure discriminant analysis and one-way ANOVA. High level of nonribosomal peptides (NRPs) only existed in the insect portions of the wild cordyceps (WI) and cultured cordyceps (CI), revealing that immunostimulation of the host insects enhanced the synthesis of NRPs in the fungus. The finding of a significantly higher level of sphingolipids in both the insect portions (WI, CI) and the coremia of the wild cordyceps (WC) and cultured cordyceps (CC) but not in cultured mycelia (CM) of I. cicadae implies that the immunostimulation of the live insects can induce the fungus to produce more sphingolipids, and this enhanced ability is probably heritable. Apart from NRPs and sphingolipids, the insect portions also contained higher levels of bioactive compounds such as lateritin, anisomycin, streptimidone and ustiloxins. In contrast, the coremium groups (WC, CC) and CM contained 10-fold less NRP but much higher levels of sanative metabolites such as tocotrienol, 3'-deoxy-hanasanagin, γ-aminobutyric acid and phospholipids than the insect portions. The significantly higher content of antioxidants in WC, CC and CM than in WI and CI suggests that environmental oxygen has a significant effect on the metabolites. The temperature stress which the wild cordyceps encounters during growth is responsible for the relatively high content of trehalose. These findings indicate that the immunity of the host insect and growth environment have a strong impact on the metabolomic variation in Isaria cicadae. The variation in metabolites suggests differential utilization value for the insect portions, coremia and mycelia of the fungus.
Assuntos
Cordyceps/química , Cordyceps/metabolismo , Metaboloma/fisiologia , Metabolômica/métodos , Micélio/química , Micélio/metabolismo , Cromatografia Líquida de Alta Pressão , Análise por Conglomerados , Cordyceps/classificação , Espectrometria de Massas , Análise de Componente Principal , Reprodutibilidade dos TestesRESUMO
A resazurin method was employed to test and compare cytotoxicity of extracts from fruiting bodies, insects and cultured mycelia of Cordyceps formosana against Chinese hamster ovary (CHO) cells. Results showed that the cultured mycelia had much stronger cytotoxicity than that of the fruiting bodies and infected insects. This suggests that using cultured mycelia to substitute a natural Cordyceps may result in poisoning. A combined method of HPLC-PAD-HRMS and cytotoxic analysis revealed that the most toxic compound (Compound 1) was found mainly in the cultured mycelia and also a small amount in the infected insect body of the Cordyceps, but not in the fruiting body. The second toxic compound (Compound 2) was found in all structures of Cordyceps and in cultured mycelia. Different contents of the toxic compounds resulted in the different cytotoxicity of the extracts. Compound 1 and Compound 2 were prepared with preparative HPLC as yellow and orange powders, respectively. Cytotoxic tests showed that the median lethal dose (LD50) against CHO cells of Compound 1 was 18.3 ± 0.2 and 103.7 ± 5.9 µg/mL for Compound 2. Compound 1 and Compound 2 were identified as rugulosin and skyrin by HRMS, UV and NMR data. The two compounds were never previously isolated from the genera Cordyceps and Hirsutella and their cytotoxicity against CHO cells was also reported for the first time.
Assuntos
Cordyceps/química , Carpóforos/química , Alimento Funcional/análise , Materia Medica/química , Micélio/química , Micotoxinas/análise , Tenebrio/química , Animais , Antraquinonas/análise , Antraquinonas/química , Antraquinonas/isolamento & purificação , Antraquinonas/toxicidade , Células CHO , China , Misturas Complexas/química , Misturas Complexas/toxicidade , Cordyceps/crescimento & desenvolvimento , Cordyceps/isolamento & purificação , Cricetinae , Cricetulus , Técnicas de Cultura , Contaminação de Medicamentos , Contaminação de Alimentos , Carpóforos/crescimento & desenvolvimento , Carpóforos/isolamento & purificação , Alimento Funcional/efeitos adversos , Hypocreales/química , Hypocreales/crescimento & desenvolvimento , Hypocreales/isolamento & purificação , Larva/química , Larva/microbiologia , Dose Letal Mediana , Materia Medica/efeitos adversos , Estrutura Molecular , Micélio/crescimento & desenvolvimento , Micélio/isolamento & purificação , Micologia/métodos , Micotoxinas/química , Micotoxinas/isolamento & purificação , Micotoxinas/toxicidade , Tenebrio/microbiologiaRESUMO
Gao-Cha is a traditional Chinese health tea made from Acer ginnala. We performed a components and radical scavenging activity analysis to identify any medicinal components in this tea. High performance thin layer chromatography (HPTLC)-1,1-Diphenyl-2-picryhydrazyl (HPTLC-DPPH) assay showed that the methanolic extract contained strong radical scavengers. Quantitative analysis revealed that the IC(50) of the extract against 1 mM DPPH was 52.7 ± 0.6 µg/mL. Bioactive-guided isolations led to procurement of 3 radical scavengers with IC(50)s of 17.5 ± 2.1, 29.3 ± 2.5, and 21.6 ± 1.7 µg/mL, respectively. Analysis of the high resolution-electric spray ionization-mass spectrometer and (1)H, (13)C, distortionless enhancement by polarization transfer at 135°, heteronuclear quantum coherence, correlating spectroscopy coupling, and heteronuclear multiple bond coherence (HMBC) data revealed that the compounds were methyl 3, 4, 5-trihydroxybenzoate (1), quercetin-3-O-α-rhamnopyranoside (2), and 2,6-bis (3,4,5-trihydroxybenzoyl)-aceritol (3). Bioactive combined components analysis revealed that, apart from compounds 1, 2, and 3, the tea possibly contained radical scavengers: ginnalin A (4) and B (5), 2â³-O-Galloylquercitrin (6) and 3â³-O-Galloyl-quercitrin (7). Compounds 2, 6, and 7 were isolated from Acer ginnala for the first time. The positions of the 2 galloyl moieties in compound 3 were unambiguously established by the HMBC spectrum for the first time.
Assuntos
Acer/química , Medicamentos de Ervas Chinesas/química , Sequestradores de Radicais Livres/química , Sequestradores de Radicais Livres/isolamento & purificação , Folhas de Planta/química , Brotos de Planta/química , Bebidas/análise , Cromatografia Líquida de Alta Pressão , Cromatografia em Camada Fina , Sequestradores de Radicais Livres/farmacologia , Ácido Gálico/análogos & derivados , Ácido Gálico/química , Ácido Gálico/isolamento & purificação , Ácido Gálico/farmacologia , Glucosídeos/química , Glucosídeos/isolamento & purificação , Glucosídeos/farmacologia , Glicosídeos , Estrutura Molecular , Monossacarídeos/química , Monossacarídeos/isolamento & purificação , Monossacarídeos/farmacologia , Ressonância Magnética Nuclear Biomolecular , Extratos Vegetais/química , Extratos Vegetais/isolamento & purificação , Extratos Vegetais/farmacologia , Quercetina/análogos & derivados , Quercetina/química , Quercetina/isolamento & purificação , Quercetina/farmacologia , Espectrometria de Massas por Ionização por ElectrosprayRESUMO
OBJECTIVE: To assess the therapeutic effect of auricular point taping and pressing therapy on vascular dementia (VD). METHODS: One hundred and eighty cases of vascular dementia were randomly divided into an auricular point taping and pressing group and a western medicine group, 90 cases in each group. The auricular point taping and pressing group was treated by auricular point taping and pressing at auricular points Shennen, Nao (brain), Shen (kidney), Zhen (pulvinal), and the western medicine group by oral administration of Nimodipine, thrice each day, 30 mg each time. They were treated for 12 weeks. The scores of mini-mental state examination (MMSE) and activities of daily living (ADL), and adverse reaction were observed. RESULTS: The scores of MMSE before treatment and 12 weeks after treatment were 18.00 +/- 3.88 and 20.83 +/- 3.74 in the auricular point taping and pressing group, and 17.80 +/- 3.82 and 20.70 +/- 3.53 in the western medicine group, respectively, with significant increases in scores of MMSE after treatment in the two groups (both P < 0.01) and with no significant difference between the two groups (P > 0.05). The scores of ADL before treatment and 12 weeks after treatment were 44.90 +/- 14.84 and 39.60 +/- 12.45 in the auricular point taping and pressing group, and 45.70 +/- 14.86 and 39.10 +/- 13.44 in the western medicine group, respectively, with significant decreases after treatment in the two groups (both P < 0.01) and with no significant difference between the two groups (P > 0.05). In the auricular point taping and pressing group, 2 cases withdrew from the test because adhesive plaster allergic reaction induced severe itch of the auricle. In the western medicine group, one case had mild dizziness and another case had diarrhea respectively. CONCLUSION: Auricular point taping and pressing and Nimodipine have similar therapeutic effect on vascular dementia, and have no obvious adverse reaction, except adhesive plaster allergic reaction.
Assuntos
Pontos de Acupuntura , Demência Vascular/tratamento farmacológico , Medicamentos de Ervas Chinesas/uso terapêutico , Manipulações Musculoesqueléticas , Idoso , Idoso de 80 Anos ou mais , Demência Vascular/terapia , Orelha Externa/efeitos dos fármacos , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Nimodipina/uso terapêuticoRESUMO
The free radical scavenging activity of the 80% methanolic extracts from fresh leaves of 300 Chinese medical woody plants was assessed with the aid of the stable DPPH radical. Among the plants screened, 56 species had strong free radical scavenging capacities, with IC50 values lower than 0.5 mg leaves per milliliter. Analysis of the medical uses of these plants showed that most of them are employed for their effects on hemostasis, as anti-inflammatory, antimicrobial or for treatment of dysentery. These uses may be directly linked to the content in tannins and flavonoids and consequently to their free radical scavenging activities.