RESUMO
Background: American ginseng (Panax quinquefolium L., AG) is a traditional Chinese medicine with multiple cardiovascular protective properties. Many bioactive components have been discovered in AG over these years. However, the understanding of these key pharmacodynamic components of activity against heart failure is insufficient. Methods: A heart failure model was established using AB line wild-type zebrafish (Danio rerio) to evaluate the anti-heart failure activity of AG. Untargeted metabolomics analysis based on ultra-high performance liquid chromatography-quadrupole electrostatic field orbitrap-mass spectrometry technology (UHPLC-QE-Orbitrap-MS) was performed to screen differential components from AG samples. The potential active components were verified using the zebrafish model. Simultaneously, network pharmacology and molecular docking techniques were used to predict the possible mechanism. Finally, the key targets of six key pharmacodynamic components were verified in zebrafish using quantitative real-time-polymerase chain reaction (Q-PCR) techniques. Results: The heart failure model was successfully established in 48 h of post-fertilization (hpf) zebrafish larvae by treating with verapamil hydrochloride. The zebrafish assay showed that the anti-heart failure effects of AG varied with producing regions. The result of the herbal metabolomic analysis based on UHPLC-QE-Orbitrap-MS indicated that ginsenoside Rg3, ginsenoside Rg5, ginsenoside Rg6, malic acid, quinic acid, L-argininosuccinic acid, 3-methyl-3-butenyl-apinosyl (1â6) glucoside, pseudoginsenoside F11, and annonaine were differential components, which might be responsible for variation in efficacy. Further analysis using zebrafish models, network pharmacology, and Q-PCR techniques showed that ginsenoside Rg3, ginsenoside Rg5, ginsenoside Rg6, malic acid, quinic acid, and pseudoginsenoside F11 were the pharmacodynamic markers (P-markers) responsible for anti-heart failure. Conclusion: We have rapidly identified the P-markers against heart failure in AG using the zebrafish model and metabolomics technology. These P-markers may provide new reference standards for quality control and new drug development of AG.
RESUMO
The rhizomes of Polygonatum odoratum represent a traditional Chinese medicine and functional food. A phytochemical investigation resulted in the isolation of eight steroidal glycosides (1-8), including two new compounds, polygonatumosides F (1) and G (2). The structures were elucidated by spectroscopic data and chemical reactions. Compound 7 showed antiproliferation activity against human hepatocellular carcinoma cell line HepG2 (IC50 of 3.2 µM). The chemical profile and contents of steroidal glycosides of P. odoratum rhizomes collected at different dates and geographical locations were also investigated, indicating that the rational harvest of P. odoratum in spring and autumn is preferable to obtain higher levels of steroidal glycosides. Compounds 1 and 7 showed the highest contents in all P. odoratum samples and have potential to serve as chemotaxonomic and chemical markers for quality control of this important plant material. 14-Hydroxylation may be a key step for the biosynthesis of compounds 1-7.
Assuntos
Glicosídeos/química , Glicosídeos/isolamento & purificação , Extratos Vegetais/química , Extratos Vegetais/isolamento & purificação , Polygonatum/química , Esteroides/química , Esteroides/isolamento & purificação , Proliferação de Células/efeitos dos fármacos , Sobrevivência Celular/efeitos dos fármacos , Cromatografia Líquida , Glicosídeos/farmacologia , Células Hep G2 , Humanos , Espectrometria de Massas , Estrutura Molecular , Extratos Vegetais/farmacologia , Rizoma/química , Esteroides/farmacologiaRESUMO
Diaphragma juglandis Fructus has been known for its effect on tonifying kidney in traditional Uighur medicine. During our ongoing studies on the discovery of biologically active compounds from folk medicine, a pair of flavanonol glycoside isomers named (2S,3S)-taxifolin-3-O-α-d-arabinofuranoside (1) and (2S,3S)-taxifolin-3-O-α-l-arabinofuranoside (2) were isolated, among them, 1 was a new compound. It is the first report of taxifolin-3-O-arabinofuranosides in both α-d and α-l configuration forms in genus Juglans. Their structures were determined on the basis of spectroscopic analysis and chemical evidence, in combination with circular dichroism.
Assuntos
Glucosídeos/química , Juglans/química , Quercetina/análogos & derivados , Dicroísmo Circular , Glucosídeos/isolamento & purificação , Glicosídeos/química , Isomerismo , Medicina Tradicional , Estrutura Molecular , Plantas Medicinais/química , Quercetina/química , Quercetina/isolamento & purificação , Espectrofotometria UltravioletaRESUMO
Although Traditional Chinese Medicine (TCM) preparations have long history with successful applications, the scientific and systematic quality assessment of TCM preparations mainly focuses on chemical constituents and is far from comprehensive. There are currently only few primitive studies on assessment of biological ingredients in TCM preparations. Here, we have proposed a method, M-TCM, for biological assessment of the quality of TCM preparations based on high-throughput sequencing and metagenomic analysis. We have tested this method on Liuwei Dihuang Wan (LDW), a TCM whose ingredients have been well-defined. Our results have shown that firstly, this method could determine the biological ingredients of LDW preparations. Secondly, the quality and stability of LDW varies significantly among different manufacturers. Thirdly, the overall quality of LDW samples is significantly affected by their biological contaminations. This novel strategy has the potential to achieve comprehensive ingredient profiling of TCM preparations.
Assuntos
DNA de Plantas/genética , Medicamentos de Ervas Chinesas/análise , Medicamentos de Ervas Chinesas/química , Sequenciamento de Nucleotídeos em Larga Escala/métodos , Algoritmos , Sequência de Bases , Contaminação de Medicamentos/prevenção & controle , Dados de Sequência MolecularRESUMO
Traditional Chinese medicine (TCM) preparations are widely used for healthcare and clinical practice. So far, the methods commonly used for quality evaluation of TCM preparations mainly focused on chemical ingredients. The biological ingredient analysis of TCM preparations is also important because TCM preparations usually contain both plant and animal ingredients, which often include some mis-identified herbal materials, adulterants or even some biological contaminants. For biological ingredient analysis, the efficiency of DNA extraction is an important factor which might affect the accuracy and reliability of identification. The component complexity in TCM preparations is high, and DNA might be destroyed or degraded in different degrees after a series of processing procedures. Therefore, it is necessary to establish an effective protocol for DNA extraction from TCM preparations. In this study, we chose a classical TCM preparation, Liuwei Dihuang Wan (LDW), as an example to develop a TCM-specific DNA extraction method. An optimized cetyl trimethyl ammonium bromide (CTAB) method (TCM-CTAB) and three commonly-used extraction kits were tested for extraction of DNA from LDW samples. Experimental results indicated that DNA with the highest purity and concentration was obtained by using TCM-CTAB. To further evaluate the different extraction methods, amplification of the second internal transcribed spacer (ITS2) and the chloroplast genome trnL intron was carried out. The results have shown that PCR amplification was successful only with template of DNA extracted by using TCM-CTAB. Moreover, we performed high-throughput 454 sequencing using DNA extracted by TCM-CTAB. Data analysis showed that 3-4 out of 6 prescribed species were detected from LDW samples, while up to 5 contaminating species were detected, suggesting TCM-CTAB method could facilitate follow-up DNA-based examination of TCM preparations.
Assuntos
DNA/isolamento & purificação , Medicamentos de Ervas Chinesas/química , Microextração em Fase Líquida/métodos , Cetrimônio , Compostos de Cetrimônio/química , DNA Intergênico/genética , Humanos , Proteínas de Plantas/genética , Reação em Cadeia da Polimerase , Reprodutibilidade dos Testes , SolventesRESUMO
Much attention has been paid to cholestane-type steroidal glycosides because of their importance from the perspectives of both chemical diversity and significant biological activities. A phytochemical investigation of the rhizomes of Polygonatum odoratum (Liliaceae) resulted in the isolation of three novel cholestane-type steroidal glycosides (1-3) with unique Δ(14,16)-unsaturated D-ring structures as well as two novel spirostane-type steroidal saponins (4 and 5) and three known steroidal glycosides (6-8). Their structures were determined by various spectroscopic methods and chemical reactions. Steroidal saponin 7 showed significant antifungal activity against Candida albicans JCM1542 (MIC 3.1 µg/mL) and Aspergillus fumigatus JCM1738 (MIC 6.3 µg/mL).
Assuntos
Colestanos/química , Glicosídeos/química , Extratos Vegetais/química , Polygonatum/química , Saponinas/química , Espirostanos/química , Colestanos/isolamento & purificação , Glicosídeos/isolamento & purificação , Estrutura Molecular , Saponinas/isolamento & purificação , Espirostanos/isolamento & purificação , EstereoisomerismoRESUMO
Homoisoflavonoid is a special type of flavonoid with the perspectives of both chemical diversity and significant biological activities. During our ongoing studies on the discovery of novel homoisoflavonoids from traditional Chinese medicines, a new minor 3,4-di-O-substituted homoisoflavonoid, 3',4-di-O-methylepisappanol (1), was isolated from Caesalpinia sappan, and its structure was determined to be (3R,4R)-3,7-dihydroxy-3-(3'-methoxy-4'-hydroxybenzyl)-4-methoxychroman by various spectroscopic analyses. Meanwhile, a known xanthone derivate, 2-methoxy-3-hydroxyxanthone (2) was also isolated, which is the first example from the genus Caesalpinia.
Assuntos
Caesalpinia/química , Medicamentos de Ervas Chinesas/isolamento & purificação , Isoflavonas/isolamento & purificação , Xantonas/isolamento & purificação , Medicamentos de Ervas Chinesas/química , Medicamentos de Ervas Chinesas/farmacologia , Flavonoides , Isoflavonas/química , Isoflavonas/farmacologia , Estrutura Molecular , Estereoisomerismo , Xantonas/química , Xantonas/farmacologiaRESUMO
The roots of Pittosporum glabratum Lindl. (Pittosporaceae) have been used as a folk medicine for the treatment of rheumatic arthritis, insomnia and hypertension. Only a few chemical or biological studies on P. glabratum have been reported. As part of our ongoing phytochemical research on this plant, four compounds were isolated. Their structures were identified as 3beta, 6beta, 19alpha, 21alpha, 24-pentahydroxy-12-en-28-oleanolic acid (1), 3-O-beta-D-glucuronopyranosyl-28-O-beta-D-glucopyranosyl siaresinolic acid (2), 3, 4, 5-trimethoxyphenyl-1-O-beta-D-(5-O-syringoyl)-apiofuranosyl-(1 --> 6)-beta-D-glucopyranoside (3) and 3, 4, 5-trimethoxyphenol-1-O-beta-D-apiofuranosyl-(1 --> 6)-beta-D-glucopyranoside (4) on the basis of physical evidence and spectroscopic analysis. Among them, compound 1 is a new triterpenoid, and compounds 2-4 are isolated from the genus Pittosporum for the first time.
Assuntos
Plantas Medicinais/química , Rosales/química , Triterpenos/isolamento & purificação , Estrutura Molecular , Ácido Oleanólico/análogos & derivados , Raízes de Plantas/química , Triterpenos/químicaRESUMO
Brazilin is a major homoisoflavonoid component isolated from the dried heartwood of traditional Chinese medicine Caesalpinia sappan L., which is a natural red pigment used for histological staining. Herein a sensitive, specific and rapid analytical LC-MS/MS method was established and validated for brazilin in rat plasma. After a simple step of protein precipitation using acetonitrile, plasma samples were analyzed using an LC-MS/MS system. Brazilin and the IS (protosappanin B) were separated on a Diamonsil C18 analytical column (150 × 4.6 mm, 5 µm) using a mixture of water and 10 mm ammonium acetate in methanol (20:80, v/v) as mobile phase at a flow rate of 0.6 mL/min. The method was sensitive with a lower limit of quantitation of 10.0 ng/mL, with good linearity (r(2) ≥ 0.99) over the linear range 10.0-5000 ng/mL. All the validation data, such as accuracy and precision, matrix effect, extraction recovery and stability tests were within the required limits. The assay method was successfully applied to evaluate the pharmacokinetics parameters of brazilin after an oral dose of 100 mg/kg brazilin in rats.
Assuntos
Benzopiranos/sangue , Benzopiranos/farmacocinética , Cromatografia Líquida de Alta Pressão/métodos , Espectrometria de Massas em Tandem/métodos , Animais , Benzopiranos/química , Estabilidade de Medicamentos , Masculino , Ratos , Ratos Sprague-Dawley , Reprodutibilidade dos Testes , Sensibilidade e EspecificidadeRESUMO
A new 19-oxo-18,19-seco-ursane-type triterpenoid saponin, named sanguisoside A (1), along with nine known triterpenoid saponins (2-10), was isolated from the roots of Sanguisorba officinalis. Their structures were determined on the basis of spectroscopic analysis and chemical method. Compounds 2 and 3 showed cytotoxic activity against SGC-7901, SMMC-7721, A549, and DU145 cell lines.
Assuntos
Antineoplásicos Fitogênicos/isolamento & purificação , Medicamentos de Ervas Chinesas/isolamento & purificação , Sanguisorba/química , Saponinas/isolamento & purificação , Triterpenos/isolamento & purificação , Antineoplásicos Fitogênicos/química , Antineoplásicos Fitogênicos/farmacologia , Ensaios de Seleção de Medicamentos Antitumorais , Medicamentos de Ervas Chinesas/química , Medicamentos de Ervas Chinesas/farmacologia , Humanos , Estrutura Molecular , Raízes de Plantas/química , Saponinas/química , Triterpenos/químicaRESUMO
OBJECTIVE: To study the lignan of Polygonum capitatum. METHODS: The compounds were isolated and purified by using of SiO2, Sephadex LH-20 column chromatography and pre-HPLC. The structures of the compounds were determined on the basis of their physical properties and the spectral data. RESULTS: Four compounds were isolated and identified as schizandriside (1), (-)-isolariciresinol-2a-O-beta-D-xylopyranoside (2), (-) -5 '-methoxyisolariciresinol-2a-O-beta-D-xylopyranoside (3) and nudiposide (4). CONCLUSION: Compounds 1 - 4 are isolated from the genus for the first time.
Assuntos
Lignanas/análise , Plantas Medicinais/química , Polygonum/química , Tetra-Hidronaftalenos/análise , Cromatografia Líquida de Alta Pressão/métodos , Lignanas/química , Lignanas/isolamento & purificação , Estrutura Molecular , Tetra-Hidronaftalenos/químicaRESUMO
A new homoisoflavan, 7,3',4'-trihydroxy-3-benzyl-2H-chromene (1), was isolated from the dried heartwood of Caesalpinia sappan L., together with seven known phenolic compounds. The structure of the new compound (1) was determined on the basis of spectroscopic analysis.