RESUMEN
The fruits of Rubus chingii Hu have high medicinal and nutritional values. However, the metabolite profiles of R. chingii, especially the alterations during different development stages of fruit, have not been comprehensively analyzed, hindering the effective utilization of the unique species. In this study, we comprehensively analyzed the metabolites of R. chingii fruit at four developmental stages using systematic untargeted and targeted liquid chromatography-mass spectrometry metabolomics analysis and identified 682 metabolites. Significant changes were observed in metabolite accumulation and composition in fruits during the different developmental stages. The contents of the index components, kaempferol-3-O-rutinoside and ellagic acid, were the highest in immature fruit. The analysis identified 64 differentially expressed flavonoids and 39 differentially expressed phenolic acids; the accumulation of most of these differentially expressed metabolites decreased with the developmental stages of fruit from immaturity to maturity. These results confirmed that the developmental stages of fruit are a critical factor in determining its secondary metabolite compositions. This study elucidated the metabolic profile of R. chingii fruit at different stages of development to understand the dynamic changes in metabolites.
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Rubus , Rubus/química , Frutas/química , Extractos Vegetales/química , Flavonoides/análisis , Cromatografía LiquidaRESUMEN
AIMS: Atractylodes macrocephala is a traditional Chinese medicine with a variety of pharmacological activities. This study aimed to evaluate its anti-hyperuricemia and antiinflammatory effects on gout, and to preliminarily explore its mechanism. METHODS: The hyperuricemia rat model was established by intraperitoneal injection of oteracil potassium and intragastric gavage of yeast powder solution. And the acute gouty arthritis (GA) model was established by injecting monosodium urate (MSU) suspension. In the study of the antihyperuricemia effect of Atractylodes macrocephala, the healthy male Sprague-Dawley rats were randomly divided into the blank group, hyperuricemia group allopurinol group as well as low, moderate and high dose groups of Atractylodes macrocephala decoction (N=8 rats in each group). Serum, liver and kidney tissue samples were collected from each group. Serum uric acid (UA), adenosine deaminase (ADA) and xanthine oxidase (XOD) levels in each group were detected by enzyme-linked immunosorbent assay (ELISA). Protein levels of ADA and XOD in liver tissues were detected by Western blot, and renal histological changes were observed by Hematoxylin-eosin (H&E) and Masson staining. In order to investigate the anti-inflammatory effect of Atractylodes macrocephala, the healthy male Sprague-Dawley rats were randomly divided into the blank group, GA group, colchicine group, high, moderate and low dose groups of Atractylodes macrocephala decoction (N=8 rats in each group), and serum and synovial tissue of each group were collected. Then the level of serum interleukin (IL)-1ß and tumor necrosis factor (TNF)-α was observed by ELISA, and the histological changes of synovial tissue were observed by H&E staining. Besides, the expression of adenosine monophosphate- activated protein kinase (AMPK) /silent information regulator (SIRT) 1/ nuclear factor kappa B (NF-κB) protein in synovial tissue was observed by Western blot and immunohistochemistry. The markers of M1 and M2 macrophages, inducible nitric oxide synthase (iNOS) and arginase-1 (ARG1) were observed by Western blot and immunofluorescence. RESULTS: Atractylodes macrocephala could reduce the production of UA by inhibiting the level of ADA and XOD, and could improve renal injury and fibrosis. In addition, Atractylodes macrophages could reduce the levels of IL-1ß and TNF-α, activate AMPK/SIRT1 signaling pathway, and inhibit the activation of NF-κB and the polarization of macrophages to a pro-inflammatory phenotype. CONCLUSION: Atractylodes macrocephala shows good anti-hyperuricemic and anti-inflammatory effects, and its anti-inflammation pharmacological activity may be related to the inhibition of M1 macrophage polarization and NF-κB activation through activating AMPK/SIRT1.
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Artritis Gotosa , Atractylodes , Hiperuricemia , Ratas , Masculino , Animales , Artritis Gotosa/inducido químicamente , Artritis Gotosa/tratamiento farmacológico , Artritis Gotosa/metabolismo , Ácido Úrico , Atractylodes/metabolismo , Sirtuina 1/uso terapéutico , FN-kappa B/metabolismo , FN-kappa B/uso terapéutico , Ratas Sprague-Dawley , Proteínas Quinasas Activadas por AMP/uso terapéutico , Hiperuricemia/tratamiento farmacológico , Antiinflamatorios/farmacología , Antiinflamatorios/uso terapéutico , Factor de Necrosis Tumoral alfa/metabolismoRESUMEN
INTRODUCTION: The root of Tetrastigma hemsleyanum (RTH) has been widely used as a folk medicine in China. Meanwhile, its stems (STH) and leaves (LTH) are consumed as functional tea and food supplementation. Therefore, it is important to get a better understanding of the distribution of bioactive constituents in different parts of T. hemsleyanum. OBJECTIVE: To develop a method for quantitative analysis of multiple bioactive constituents and comparing their distribution in RTH, STH and LTH. METHODS: Ultra-performance liquid chromatography triple quadrupole ion trap tandem mass spectrometry (UPLC-QTRAP-MS/MS) was used for the quantitative analysis. The quantitative data were further analysed by principal component analysis (PCA), hierarchical cluster analysis (HCA) and partial least squares determinant analysis (PLS-DA). RESULTS: Forty-two constituents in RTH, STH and LTH, including 14 flavonoids, three phenolic acids, 15 amino acids and 10 nucleosides, were quantitatively determined. The contents of flavonoids and phenolic acids in LTH were significantly higher than those in RTH and STH. While the contents of amino acids and nucleosides in LTH were less than those in RTH and STH. Multivariate statistical analysis can significantly classify and distinguish RTH, STH, and LTH. CONCLUSIONS: The present method would be helpful for the quality control of T. hemsleyanum, and the results would be useful for the efficient utilisation of T. hemsleyanum in the future.
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Espectrometría de Masas en Tándem , Vitaceae , Aminoácidos , Cromatografía Líquida de Alta Presión/métodos , Flavonoides/química , Espectrometría de Masas en Tándem/métodos , Vitaceae/químicaRESUMEN
BACKGROUND: Bletilla striata is the main medicine of many skin whitening classic formulas in traditional Chinese medicine (TCM) and is widely used in cosmetic industry recently. However, its active ingredients are still unclear and its fibrous roots are not used effectively. The aim of the present study is to discover and identify its potential anti-melanogenic active constituents by zebrafish model and molecular docking. METHODS: The antioxidant activities were evaluated by 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical scavenging activity, 2,2'-azino-bis-(3-ethylbenthiazoline-6-sulphonic acid) (ABTS) radical scavenging activity and ferric reducing antioxidant power (FRAP) assay. The anti-melanogenic activity was assessed by tyrosinase inhibitory activity in vitro and melanin inhibitory in zebrafish. The chemical profiles were performed by ultra-high-performance liquid chromatography combined with quadrupole time-of-flight tandem mass spectrometry (UPLC-Q-TOF-MS/MS). Meanwhile, the potential anti-melanogenic active constituents were temporary identified by molecular docking. RESULTS: The 95% ethanol extract of B. striata fibrous roots (EFB) possessed the strongest DPPH, ABTS, FRAP and tyrosinase inhibitory activities, with IC50 5.94 mg/L, 11.69 mg/L, 6.92 mmol FeSO4/g, and 58.92 mg/L, respectively. In addition, EFB and 95% ethanol extract of B. striata tuber (ETB) significantly reduced the melanin synthesis of zebrafish embryos in a dose-dependent manner. 39 chemical compositions, including 24 stilbenoids were tentatively identified from EFB and ETB. Molecular docking indicated that there were 83 (including 60 stilbenoids) and 85 (including 70 stilbenoids) compounds exhibited stronger binding affinities toward tyrosinase and adenylate cyclase. CONCLUSION: The present findings supported the rationale for the use of EFB and ETB as natural skin-whitening agents in pharmaceutical and cosmetic industries.
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Antioxidantes/farmacología , Medicina Tradicional China/métodos , Melaninas/antagonistas & inhibidores , Simulación del Acoplamiento Molecular , Extractos Vegetales/farmacología , Polisacáridos/farmacología , Preparaciones para Aclaramiento de la Piel/farmacología , Animales , Antioxidantes/química , China , Modelos Animales , Monofenol Monooxigenasa/antagonistas & inhibidores , Extractos Vegetales/química , Raíces de Plantas , Tubérculos de la Planta , Polisacáridos/química , Preparaciones para Aclaramiento de la Piel/química , Pez CebraRESUMEN
Saururus chinensis (SC) possesses significant anti-diabetic activity and lignans were its major bioactive compounds. In this study, a rapid and sensitive ultra-high-performance liquid chromatography-tandem mass spectrometry (UHPLC-MS/MS) method was established for simultaneous quantification of six lignans, namely (-)-(7R,8R)-machilin D (1), verrucesin (2), rel-(7S,8S,7'R,8'R)-3,3',4,4',5,5'-hexamethoxy-7.O.7',8.8'-lignan (3), manassantin A (4), manassantin B (5), and saucerneol F (6) in rat's plasma. It was validated with acceptable linearity (r ≥ 0.9922), accuracy (80.42-95.17%), precision (RSD ≤ 12.08%), and extraction recovery (80.36-93.45%). The method was successfully applied to the comparative pharmacokinetic study of the six lignans in normal and diabetic rats after oral administration of SC extract. Results showed that the areas under the plasma concentration-time curve (AUC0 â t and AUC0 â ∞ ) of (-)-(7R,8R)-machilin D, rel-(7S,8S,7'R,8'R)-3,3',4,4',5,5'-hexamethoxy-7.O.7',8.8'-lignan, manassantin B, and saucerneol F in diabetic rats were significantly increased, and the plasma clearance (CL) of (-)-(7R,8R)-machilin D in diabetic rats was significantly decreased. However, the AUC0 â t and AUC0 â ∞ of verrucesin were significantly decreased, and its CL was significantly increased in diabetic rats compared with those in normal rats. These results indicated that there were remarkable differences in the pharmacokinetic parameters between the normal and diabetic rats. The pharmacokinetic studies might be beneficial for the clinical use of SC as hypoglycemic agent.
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Diabetes Mellitus Experimental/metabolismo , Lignanos , Extractos Vegetales , Saururaceae/química , Administración Oral , Animales , Cromatografía Líquida de Alta Presión/métodos , Lignanos/sangre , Lignanos/química , Lignanos/farmacocinética , Masculino , Extractos Vegetales/administración & dosificación , Extractos Vegetales/farmacocinética , Ratas , Ratas Sprague-Dawley , Espectrometría de Masas en Tándem/métodosRESUMEN
Houttuynia cordata has been used as a traditional medicine for more than 1500 years. It has aroused wide public concern about its safety in the past few years, for it contains various aristolactams. However, the safety of H. cordata extract remains unclear. In the present study, single dose (2000 mg/kg) and subacute (250, 500, and 1000 mg/kg/day for 28 days) oral toxicity studies of the 95% ethanol extract of H. cordata (HCE) were performed in both male and female Sprague-Dawley (SD) rats. Hematological, biochemical, histopathological parameters, and plasma metabolic profiling were assessed. The single-dose toxicity of HCE was more than 2000 mg/kg. The subacute toxicity results showed that no significant adverse effect of HCE was observed at 250 mg/kg/day. However, five rats died in 500 and 1000 mg/kg/day groups and exhibited toxicities to liver and kidney. Plasma metabolic profiling analysis suggested that a number of metabolic disturbances were induced by oral administration of HCE, focusing on energy metabolism, amino acid metabolism, and lipids metabolism. Moreover, it appeared that male rats were more susceptible to the toxic effects of HCE than female rats. Therefore, in this preliminary study, oral administration of HCE 250 mg/kg/day can be regarded as the no observed adverse effect level in rats over 28 days. However, long-term use of HCE with large doses exhibited some hepatotoxicity and nephrotoxicity in rats.
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Houttuynia/química , Metaboloma/efectos de los fármacos , Extractos Vegetales/toxicidad , Animales , Femenino , Masculino , Extractos Vegetales/química , Ratas , Ratas Sprague-Dawley , Pruebas de Toxicidad Aguda , Pruebas de Toxicidad SubagudaRESUMEN
ETHNOPHARMACOLOGICAL RELEVANCE: Fritillariae Thunbergii Flos (FTF) included in the Chinese Pharmacopoeia (1977 Edition) is a Chinese medicinal herb traditionally used to treat bronchitis. In recent years, it has been applied in the treatment of lung cancer. However, the molecular mechanism remains largely unknown. METHODS: The screening of bioactive compounds, acquisition of drug targets, network construction, and experimental validation in vivo were combined to explored the mechanism of FTF in the treatment of lung carcinoma with regards to systems pharmacology. RESULTS: The network Lung Cancer Pathway consisted of 114 nodes (44 compounds and 70 potential targets) and 361 edges, as well as modules that included inflammatory response, angiogenesis, negative regulation of the apoptotic process, and positive regulation of cell proliferation and migration. It was examined by conducting experiments that involved the administration of ethanol-based extracts of FTF in Lewis lung carcinoma mice. The extracts exerted excellent anti-lung cancer effects in vivo by significantly inhibiting tumor proliferation, thereby extending the survival period of tumor-bearing mice. Moreover, FTF induced the downregulation of PIK3CG, Bcl-2, eNOS, VEGF, p-STAT3, and STAT3 genes in tumor-bearing mice. CONCLUSIONS: The findings of the present study verify the therapeutic effects and mechanism of FTF on lung cancer and provide a theoretical basis to support the comprehensive utilization of FTF resources.
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Antineoplásicos Fitogénicos/uso terapéutico , Medicamentos Herbarios Chinos/uso terapéutico , Fritillaria , Neoplasias Pulmonares/tratamiento farmacológico , Mapas de Interacción de Proteínas/efectos de los fármacos , Animales , Antineoplásicos Fitogénicos/aislamiento & purificación , Antineoplásicos Fitogénicos/farmacología , Carcinoma Pulmonar de Lewis/tratamiento farmacológico , Carcinoma Pulmonar de Lewis/genética , Carcinoma Pulmonar de Lewis/patología , Ensayos de Selección de Medicamentos Antitumorales/métodos , Medicamentos Herbarios Chinos/aislamiento & purificación , Medicamentos Herbarios Chinos/farmacología , Fritillaria/genética , Neoplasias Pulmonares/genética , Neoplasias Pulmonares/patología , Ratones , Ratones Endogámicos C57BL , Mapas de Interacción de Proteínas/fisiología , Distribución Aleatoria , Resultado del Tratamiento , Carga Tumoral/efectos de los fármacos , Carga Tumoral/fisiologíaRESUMEN
INTRODUCTION: Lindera aggregata is a main Chinese herb of ancient prescriptions Suoquan pill applied for treating the chronic kidney disease (CKD). A large number of application histories of Lindera aggregata in the treatment of CKD have been recorded in Chinese traditional medical literature. The previous reports revealed that Lindera aggregata can treat CKD. METHODS: Rats were randomly divided into control, model, Huangkui,Lindera aggregata ethanol extract (LEE) and Lindera aggregata water extract (LWE) groups. hematoxylin-eosin (HE) staining was used to detect the pathology of kidney. The levels of serum creatinine (Scr), serum Neutrophil gelatinase-associated lipocalin (NGAL), blood urea nitrogen (BUN), urine protein (UP), kidney index(KI) were evaluated. The UPLC - QTOF/MS were applied to probe the metabolic profile. Furthermore, Indoxyl sulfate-induced human renal tubular epithelial (HK-2) cell model was built to determine the expression levels of pathogenesis-related proteins. RESULTS: The results demonstrated that LEE and LWE significantly inhibited the rebound in Scr, BUN, NGAL, UP and KI in models, except for the effect of LWE at low dose (LWE-L) and LEE at low dose (LEE-L) on KI and the effect of LWE-H at high dose (LWE-H) and LEE-L on BUN and NGAL. Moreover,Lindera aggregata extracts alleviated renal tubular dilatation, interstitial fibrosis and interstitial inflammation. By analysis, twenty-eight metabolites were related to CKD. After intervention of Lindera aggregata extracts, some metabolites approach to a normal-like level, such as Indoxyl sulfate. These metabolites are mainly involved in tryptophan, fatty acid, glycerophospholipid, tyrosine and arachidonic acid metabolic pathways. Furthermore, Lindera aggregata extracts mediate the expression of smad2, smad3, smad7 and TGF-ß in Indoxyl sulfate-induced HK-2 cell. CONCLUSIONS: Lindera aggregata extracts can mitigate adenine-induced CKD by modulating the metabolic profile and TGF-ß/Smad signaling pathway, providing important supports for developing protective agent of Lindera aggregata for CKD.
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Adenina/metabolismo , Medicamentos Herbarios Chinos/farmacología , Lindera/química , Insuficiencia Renal Crónica/tratamiento farmacológico , Proteínas Smad/metabolismo , Factor de Crecimiento Transformador beta/metabolismo , Animales , Cromatografía Líquida de Alta Presión/métodos , Fibrosis/patología , Humanos , Riñón/efectos de los fármacos , Pruebas de Función Renal , Masculino , Espectrometría de Masas/métodos , Metabolómica , Fitoterapia/métodos , Ratas , Insuficiencia Renal Crónica/patología , Transducción de Señal/efectos de los fármacosRESUMEN
Scrophulariae Radix is one of the most popular traditional Chinese medicines (TCMs). Primary processing of Scrophulariae Radix is an important link which closely related to the quality of products in this TCM. The aim of this study is to explore the influence of different processing methods on chemical constituents in Scrophulariae Radix. The difference of chemical constituents in Scrophulariae Radix processed by different methods was analyzed by using ultra fast liquid chromatography-triple quadrupole-time of flight mass spectrometry coupled with principal component analysis and orthogonal partial least squares discriminant analysis. Furthermore, the contents of 12 index differential constituents in Scrophulariae Radix processed by different methods were simultaneously determined by using ultra fast liquid chromatography coupled with triple quadrupole-linear ion trap mass spectrometry. Gray relational analysis was performed to evaluate the different processed samples according to the contents of 12 constituents. All of the results demonstrated that the quality of Scrophulariae Radix processed by "sweating" method was better. This study will provide the basic information for revealing the change law of chemical constituents in Scrophulariae Radix processed by different methods and facilitating selection of the suitable processing method of this TCM.
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Cromatografía Líquida de Alta Presión/métodos , Medicamentos Herbarios Chinos/análisis , Medicamentos Herbarios Chinos/química , Scrophularia , Espectrometría de Masas en Tándem/métodos , Ácidos Carbocíclicos/análisis , Glucósidos/análisis , Límite de Detección , Modelos Lineales , Fenoles/análisis , Análisis de Componente Principal , Reproducibilidad de los ResultadosRESUMEN
Scrophulariae Radix is one of the most popular traditional Chinese medicines. The harvesting time of Scrophulariae Radix is closely related to the quality of products in this traditional Chinese medicine. The goal of the study is to analyze the dynamic changes of metabolite accumulation in Scrophulariae Radix. The difference of constituents in Scrophulariae Radix harvested at different times was analyzed by liquid chromatography with triple quadrupole-time of flight tandem mass spectrometry coupled with principal component analysis and partial least-square discriminant analysis. According to the accurate mass of molecular and product ions provided by triple quadrupole-time of flight tandem mass spectrometry, a total of 30 differential constituents were identified. Furthermore, the contents of ten index differential constituents in Scrophulariae Radix were simultaneously determined by liquid chromatography coupled with triple quadrupole-linear ion trap tandem mass spectrometry. Gray relational analysis was performed to evaluate the samples harvested at different times according to the contents of ten constituents. All of the results demonstrated that the quality of Scrophulariae Radix collected at traditional harvest time was better. This study will provide the basic information for revealing the dynamic change law of metabolite accumulation of Scrophulariae Radix and exploring its quality forming mechanism.
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Cromatografía Líquida de Alta Presión , Medicamentos Herbarios Chinos/química , Scrophularia/química , Espectrometría de Masas en Tándem , Raíces de Plantas/químicaRESUMEN
Pseudostellariae Radix (PR) is an important traditional Chinese medicine (TCM), which is consumed commonly for its positive health effects. However, the chemical differences of PR from different cultivated fields and germplasms are still unknown. In order to comprehensively compare the chemical compositions of PR from different cultivated fields, in this study, ¹H-NMR-based metabolomics coupled with high performance liquid chromatography (HPLC) were used to investigate the different metabolites in PR from five germplasms (jr, zs1, zs2, sb, and xc) cultivated in traditional fields (Jurong, Jiangsu, JSJR) and cultivated fields (Zherong, Fujian, FJZR). A total of 34 metabolites were identified based on ¹H-NMR data, and fourteen of them were found to be different in PR from JSJR and FJZR. The relative contents of alanine, lactate, lysine, taurine, sucrose, tyrosine, linolenic acid, γ-aminobutyrate, and hyperoside in PR from JSJR were higher than that in PR from FJZR, while PR from FJZR contained higher levels of glutamine, raffinose, xylose, unsaturated fatty acid, and formic acid. The contents of Heterophyllin A and Heterophyllin B were higher in PR from FJZR. This study will provide the basic information for exploring the influence law of ecological environment and germplasm genetic variation on metabolite biosynthesis of PR and its quality formation mechanism.
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Espectroscopía de Resonancia Magnética , Metaboloma , Metabolómica , Raíces de Plantas/química , Tracheophyta/química , Medicamentos Herbarios Chinos/química , Espectroscopía de Resonancia Magnética/métodos , Metabolómica/métodos , Raíces de Plantas/metabolismo , Espectroscopía de Protones por Resonancia Magnética , Tracheophyta/metabolismoRESUMEN
Scrophulariae Radix is one of the most popular traditional Chinese medicines (TCMs), which needs to be processed by 'sweating' methods. Primary processing of Scrophulariae Radix is an important link which closely relates to the quality of products in this TCM. To facilitate selection of the suitable 'sweating' processing method for Scrophulariae Radix, in this study the quality of Scrophulariae Radix processed by different 'sweating' methods was evaluated based on simultaneous determination of multiple bioactive constituents combined with grey relational analysis. The contents of iridoid glycosides, phenylpropanoid glycosides, and organic acids in Scrophulariae Radix processed by different 'sweating' methods were simultaneously determined using ultra high performance liquid chromatography coupled with triple quadrupole-linear ion trap mass spectrometry (UPLC-QTRAP-MS/MS). Furthermore, grey relational analysis (GRA) was performed to evaluate the 'sweating' processed samples according to the contents of twelve constituents. All of the results demonstrated that the quality of Scrophulariae Radix processed by oven drying at 35 °C and 'sweating' for three days was better. The developed method was useful for the overall assessment on quality of Scrophulariae Radix, and this study may provide the foundation and support for 'sweating' processing of Scrophulariae Radix in normalization and standardization.
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Medicamentos Herbarios Chinos/química , Scrophularia/química , Cromatografía Líquida de Alta Presión , Estabilidad de Medicamentos , Límite de Detección , Estructura Molecular , Reproducibilidad de los Resultados , Espectrometría de Masas en TándemRESUMEN
Pseudostellariae Radix is an important traditional Chinese medicine (TCM), which is consumed commonly for its positive health effects. However, a lack of transcriptomic and genomic information hinders research on Pseudostellariae Radix. Here, high-throughput RNA sequencing (RNA-seq) was employed for the de novo assembly to analyze the transcriptome in Pseudostellariae Radix, finding significantly differentially expressed genes in this TCM from different fields based on RNA-seq and bioinformatic analysis. A total of 146,408,539 paired-end reads were generated and assembled into 89,857 unigenes with an average length of 862bp. All of the assembly unigenes were annotated by running BLASTx and BLASTn similarity searches on the Non-redundant nucleotide database (NT), the Non-redundant protein database (NR), Swiss-Prot, Cluster of Orthologous Groups (COG), Kyoto Encyclopedia of Genes and Genomes (KEGG), Gene Ontology (GO), and Interpro. On the basis of bioinformatic analysis and the expression profiles for Pseudostellariae Radix, 29 significantly differentially expressed genes were identified, which provides the basic information for exploring the molecular mechanisms that determine the quality of Pseudostellariae Radix from different fields. The expression levels of 29 genes were validated by real-time quantitative PCR (RT-qPCR). This is the first study to sample Pseudostellariae Radix, which provides an invaluable resource for understanding the genome of this herb.
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Caryophyllaceae/genética , Caryophyllaceae/metabolismo , China , Perfilación de la Expresión Génica , Secuenciación de Nucleótidos de Alto Rendimiento , Raíces de Plantas/genética , Raíces de Plantas/metabolismo , Reacción en Cadena en Tiempo Real de la Polimerasa , Análisis de Secuencia de ARNRESUMEN
Pseudostellaria heterophylla is an important traditional Chinese herbal medicine with vast clinical consumption because of its positive effects. To date, changes in the metabolite composition of P. heterophylla have been well documented; however, the molecular differences between cultivated P. heterophylla and its wild-type are still unknown. The aim of this study was to investigate differences in cultivated and wild P. heterophylla. Due to the lack of a genomic database, we used high-throughput transcriptomic and proteomic technologies to identify proteins in the herb. Isobaric tags for relative and absolute quantification (iTRAQ) MS/MS were used to detect statistically significant changes between cultivated and wild P. heterophylla. We detected 3775 proteins; 332 showed differential accumulations across two different ecotypes of P. heterophylla. 71 significant differential expressions of proteins were selected based on GO annotations, KEGG, STRING analysis, and expression level. Quantitative real-time PCR analysis confirmed the authenticity and accuracy of the proteomic analysis. The results indicated that the carbohydrate and cellular amino acid metabolisms in cultivated P. heterophylla were weaker than those in its wild-type; seven important proteins were found to regulate sucrose and amino acids. This will provide the basic information for exploring the cause of differences in secondary metabolites in different ecotypes of P. heterophylla and the protein mechanism of its quality formation. BIOLOGICAL SIGNIFICANCE: This study combined a transcriptome, proteome, and metabolism approach for analyzing differentially expressed proteins of cultivated and wild P. heterophylla and established the relationship between significantly differentially expressed proteins and differential chemical components in non-model plants. The results of proteomic analysis provide the basic information for exploring the quality forming process, which will demonstrate, and provide guidance for, the study of effective constituents of P. heterophylla and its quality formation mechanism.
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Caryophyllaceae/metabolismo , Regulación de la Expresión Génica de las Plantas/fisiología , Proteínas de Plantas/biosíntesis , Proteoma/biosíntesis , Caryophyllaceae/genética , Proteínas de Plantas/genética , Proteoma/genética , ProteómicaRESUMEN
To explore rapidly the potential chemical markers for differentiating Pseudostellariae Radix from different cultivated fields :and germplasms, a method is proposed based on ultra-performance liquid chromatographytriple time-of-flight mass/mass spectrometry (UPLC-Triple TOF-MS/MS) coupled with multivariate statistical analysis. Peak matching, peak alignment, and noise filtering were used in analyzing mass spectrometric data. Accurate m/z value analysis of MS data based on software of database search and MS/MS fragment analysis were applied to identify constituents. The obtained. data were statistically analyzed with hierarchical cluster analysis (HCA), principal component analysis (PCA), and partial least squared-discriminant analysis (PLS-DA) to compare the differences among these samples. The PLS-DA loading plot obtained from all mass data showed that 21 compounds were identified as the potential chemical markers to characterize the samples. The results provide experimental data to reveal the influence of ecological environments and germplasms on metabolite biosynthesis in Pseudostellariae Radix.
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Caryophyllaceae/química , Fitoquímicos/análisis , Cromatografía Líquida de Alta Presión , Análisis Multivariante , Extractos Vegetales/química , Espectrometría de Masas en Tándem , Tiempo (Meteorología)RESUMEN
A comprehensive analytical method based on UPLC-MS/MS was developed for the simultaneous determination of thirteen components including three stilbenes (stilbeneglucoside, polydatin, resveratrol), four anthraquinones (emodin, physcion, emodin-8-ß-D-glucopyranoside, aloe-emodin), five flavonoids (epicatechin, rutin, hyperoside, astragalin,quercetin) and one phenolic acid (gallic acid) in Polygoni Multifori Caulis.The separation was carried out on a Waters BEH C18 columnï¼2.1 mm×100 mm,1.7 µmï¼with gradient elution of acetonitrile-water (0.1% acetic acid) at a flow rate of 0.25 mLâ¢min⻹, and column temperature was 35 â. The target compounds were analyzed by multiple reaction monitoring (MRM) mode. TOPSIS analysis ware performed to evaluate the samples from different areas and commercial herbs according to the contents of thirteen components. The correlation coefficients of all the calibration curves were higher than 0.991 5. The average recoveries ranged from 95.24% to 102.3%, and the relative standard deviations were less than 5%. The result of TOPSIS analysis showed that the comprehensive quality of Polygoni Multifori Caulis sample from Guangzhou was better. The developed method with good repeatability and accuracy was suitable for the simultaneous determination of multiple functional substances, which provided a new basis for the comprehensive assessment and overall control of the quality of Polygoni Multifori Caulis.
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Fitoquímicos/análisis , Polygonum/química , Cromatografía Líquida de Alta Presión , Flavonoides/análisis , Ácido Gálico/análisis , Tallos de la Planta/química , Estilbenos/análisis , Espectrometría de Masas en TándemRESUMEN
To study the dynamic change law of bioactive constituents from Polygonum multiflorum, and to explore the optimal harvest period of P. multiflorum. Determination of stilhene glucoside, anthraquinones and catechin from P. multiflorum in different harvest times by MEKC-DAD, and principal component analysis (PCA) was used to comprehensive evaluation for bioactive constituents. There are obvious differences among the contents of active ingredients in various collecting periods samples, the content of stilbene glucoside was the highest in November, the total content of combined anthraquinone was the highest in November and December, the content of catechin was the highest in September. The comprehensive evaluation index obtained with principal component analysis showed that the sample collected in November is significantly higher than those with other samples. The optimal harvest period of P. multiflorum is November.
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Fallopia multiflora/química , Electroforesis , Fallopia multiflora/crecimiento & desarrollo , Fallopia multiflora/metabolismo , Factores de TiempoRESUMEN
OBJECTIVE: To study the chemical constituents of fermented Antrodia camphorata powder. METHODS: 15 compounds were isolated from Antrodia camphorata by Silica gel column chromatography, ODS column chromatography, gel column chromatography, preparative liquid phase chromatography separation technique, as well as recrystallization. RESULTS: On the basis of their physical and chemical properties and spectral data,their structures were identified as Ferulic acid (1), Inositol (2), ß-Sitosterol (3),Vanillin (4),Vanillic acid (5), Butyric acid (6), Daucosterol (7), p-Hydroxycinnamic acid (8), Lauric acid (9), Inosine (10), Uridine (11), Adenine (12), D(+)-Sucrose (13), Arachidic acid (14) and Guanosine (15). CONCLUSION: Compounds 1, 5, 6 and 8-15 are isolated from fermented powder for the first time.
Asunto(s)
Antrodia/química , Fitoquímicos/química , Plantas Medicinales/química , Cromatografía Liquida , Ácidos Cumáricos , Fermentación , Fitoquímicos/aislamiento & purificación , Polvos , Propionatos , Sitoesteroles , Ácido VanílicoRESUMEN
OBJECTIVE: To analyze the dynamic changes of nucleosides and nucleobases in Polygoni Multiflori Radix harvested in different periods. METHODS: UPLC-QTRAP-MS/MS method was applied for the analysis of nine kinds of nucleosides and nucleobases in Polygoni Multiflori Radix. RESULTS: The content of uridine, adenine, guanosine and cytidine was higher in Polygoni Multiflori Radix harvested in different periods and assumed some difference. The trends of nucleosides and nucleobases from Polygoni Multiflori Radix according to the peak valley shape changed. The highest contents of them were in December. CONCLUSION: The accumulation of nucleosides and nucleobases in Polygoni Multiflori Radix is closely related to its growth cycle. It is found to be basically the same as that obtained when the herb is collected during the conventional collecting time.