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Ulcerative colitis is closely associated with the dysregulation of gut microbiota. There is growing evidence that natural products may improve ulcerative colitis by regulating the gut microbiota. In this research, we demonstrated that bergenin, a naturally occurring isocoumarin, significantly ameliorates colitis symptoms in dextran sulfate sodium (DSS)-induced mice. Transcriptomic analysis and Caco-2 cell assays revealed that bergenin could ameliorate ulcerative colitis by inhibiting TLR4 and regulating NF-κB and mTOR phosphorylation. 16S rRNA sequencing and metabolomics analyses revealed that bergenin could improve gut microbiota dysbiosis by decreasing branched-chain amino acid (BCAA) levels. BCAA intervention mediated the mTOR/p70S6K signaling pathway to exacerbate the symptoms of ulcerative colitis in mice. Notably, bergenin greatly decreased the symbiotic bacteria Bacteroides vulgatus (B. vulgatus), and the gavage of B. vulgatus increased BCAA concentrations and aggravated the symptoms of ulcerative colitis in mice. Our findings suggest that gut microbiota-mediated BCAA metabolism plays a vital role in the protective effect of bergenin on ulcerative colitis, providing novel insights for ulcerative colitis prevention through manipulation of the gut microbiota.
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Bacteroides , Benzopiranos , Colitis Ulcerosa , Colitis , Animales , Ratones , Humanos , Colitis Ulcerosa/inducido químicamente , Colitis Ulcerosa/tratamiento farmacológico , Células CACO-2 , ARN Ribosómico 16S , Colitis/inducido químicamente , Colitis/tratamiento farmacológico , Aminoácidos de Cadena Ramificada , Serina-Treonina Quinasas TOR/genética , Sulfato de Dextran/efectos adversos , Modelos Animales de Enfermedad , Ratones Endogámicos C57BL , ColonRESUMEN
Gut microbiota dysbiosis is associated with the development of rheumatoid arthritis (RA). Sinomenine (SIN) is an effective immunosuppressive and anti-inflammatory drug used for treating RA, but how SIN regulates gut microbiota to alleviate RA remains underexplored. To identify the critical gut microbial species and microbial metabolites associated with the RA-protective effects of SIN, the microbiota-dependent anti-RA effects of SIN were assessed by 16S rRNA gene sequencing, antibiotic treatment, and fecal microbiota transplantation. Metabolomics analysis, transcriptional analysis, and targeted bacteria/metabolites gavage were conducted to explore how SIN regulates gut microbiota to reduce the severity of RA. SIN could restore intestinal microbial balance by mainly modulating the abundance of Lactobacillus, and significantly relieve collagen-induced arthritis (CIA) symptoms in a gut microbiota-dependent manner. SIN significantly elevated microbial tryptophan metabolites indole-3-acrylic acid (IA), indole-3-propionic acid (IPA), and indole-3-acetic acid (IAA). Tryptophan metabolites supplementation could activate aryl hydrocarbon receptor (AhR) and regulate Th17/Treg balance in CIA rats. Intriguingly, SIN relieved the arthritis symptoms involving the enrichment of two beneficial anti-CIA Lactobacillus species, L. paracasei and L. casei by mono-colonization. The promising therapeutic function of SIN was mostly attributed to the activation of AhR by explicitly targeting the Lactobacillus and microbial tryptophan metabolites. The intestinal bacterium L. paracasei and L. casei may be used to reduce the severity of CIA.
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The link between gut microbiota and obesity or other metabolic syndromes is growing increasingly clear. Natural products are appreciated for their beneficial health effects in humans. Increasing investigations demonstrated that the anti-obesity bioactivities of many natural products are gut microbiota dependent. In this review, we summarized the current knowledge on anti-obesity natural products acting through gut microbiota according to their chemical structures and signaling metabolites. Manipulation of the gut microbiota by natural products may serve as a potential therapeutic strategy to prevent obesity.
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Productos Biológicos , Microbioma Gastrointestinal , Síndrome Metabólico , Humanos , ObesidadRESUMEN
Metabolic syndrome (MetS) is intricately linked to dysregulation of gut microbiota and host metabolomes. Here, we first find that a purified citrus polymethoxyflavone-rich extract (PMFE) potently ameliorates high-fat diet (HFD)-induced MetS, alleviates gut dysbiosis, and regulates branched-chain amino acid (BCAA) metabolism using 16S rDNA amplicon sequencing and metabolomic profiling. The metabolic protective effects of PMFE are gut microbiota dependent, as demonstrated by antibiotic treatment and fecal microbiome transplantation (FMT). The modulation of gut microbiota altered BCAA levels in the host serum and feces, which were significantly associated with metabolic features and actively responsive to therapeutic interventions with PMFE. Notably, PMFE greatly enriched the commensal bacterium Bacteroides ovatus, and gavage with B. ovatus reduced BCAA concentrations and alleviated MetS in HFD mice. PMFE may be used as a prebiotic agent to attenuate MetS, and target-specific microbial species may have unique therapeutic promise for metabolic diseases.
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Flavonas/farmacología , Microbioma Gastrointestinal/efectos de los fármacos , Síndrome Metabólico/tratamiento farmacológico , Obesidad/tratamiento farmacológico , Aminoácidos/efectos de los fármacos , Aminoácidos/metabolismo , Animales , Citrus/química , Disbiosis/tratamiento farmacológico , Disbiosis/microbiología , Disbiosis/patología , Heces/microbiología , Flavonas/química , Humanos , Resistencia a la Insulina/genética , Síndrome Metabólico/microbiología , Síndrome Metabólico/patología , Metaboloma/efectos de los fármacos , Ratones , Obesidad/metabolismo , Obesidad/microbiología , Obesidad/patología , Prebióticos/microbiologíaRESUMEN
The traditional Chinese medicine Citri Reticulatae Pericarpium (CRP) was mainly originated from the dried pericarp of Citrus reticulata 'Chachi' (Crc), Citrus reticulata 'Dahongpao' (Crd), Citrus reticulata 'Unshiu' (Cru) and Citrus reticulata 'Tangerina' (Crt) in China. Since these four cultivars have great similarities in morphology, reliable methods to differentiate CRP cultivars have rarely been reported. To discriminate the differences of these CRP cultivars, herein an efficient and reliable method by combining metabolomics, DNA barcoding and electronic nose was first established. The hierarchical three-step filtering metabolomics analysis based on liquid chromatography-quadrupole time-of-flight mass spectrometry (LC-QTOF-MS) indicated that 9 species-specific chemical markers including 6 flavanone glycosides and 3 polymethoxyflavones could be considered as marker metabolites for discrimination of the geoherb Crc from other cultivars. A total of 19 single nucleotide polymorphism (SNP) sites were found in nuclear internal transcribed spacer 2 (ITS2) of CRP, and three stable SNP sites (33, 128 and 174) in the ITS2 region can distinguish the four CRP cultivars. The electronic nose coupled with chemometrics could also be used to effectively distinguish Crc from other CRP cultivars. Therefore, our results indicated that the integrated method will be an effective strategy for discrimination of similar herbal medicines.
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Citrus/clasificación , Código de Barras del ADN Taxonómico , Nariz Electrónica , Metabolómica , Citrus/genética , Citrus/crecimiento & desarrollo , Citrus/metabolismo , ADN Intergénico/genéticaRESUMEN
Background: Aurantii Fructus Immaturus (AFI) and Aurantii Fructus (AF) are two traditional citrus herbs with health-promoting and nutritive properties. Objective: This paper presents the first attempt to simultaneously investigate the absorption of five major flavanone glycosides, namely narirutin, naringin, hesperidin, neohesperidin, and poncirin, in rat plasma following a single oral administration of AFI and AF extracts to rats. Methods: The plasma concentrations were determined by liquid-liquid extraction followed by a rapid and sensitive ultra-performance LC-tandem mass spectrometry method. Pharmacokinetic parameters were analyzed by noncompartmental modeling using DAS software. Results: The developed method was validated and successfully applied to the pharmacokinetic study of these five flavanone glycosides. Conclusions: The comparison of the pharmacokinetic parameters of flavanone glycosides showed that the absorption of AF extract was lower, while the elimination was relatively rapid, compared with those of AFI extract. Highlights: This study may be useful for further utilization of these two citrus herbs.
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Cromatografía Líquida de Alta Presión/métodos , Citrus/química , Medicamentos Herbarios Chinos/farmacocinética , Flavanonas/sangre , Glicósidos/sangre , Espectrometría de Masas en Tándem/métodos , Administración Oral , Animales , Medicamentos Herbarios Chinos/administración & dosificación , Flavanonas/farmacocinética , Glicósidos/farmacocinética , Masculino , Ratas Sprague-DawleyRESUMEN
BACKGROUND: Inhibition of pancreatic lipase is an attractive approach to the treatment of obesity and other metabolic disorders. Naturally occurring phytochemicals are promising sources of lipase inhibitors. PURPOSE: In the present study, the anti-lipase activity of Citri Reticulatae Pericarpium (CRP) extracts was firstly evaluated in vitro. Moreover, the dynamic alteration of bioactive flavonoids in CRP collected at different time and its correlation with anti-lipase activities was investigated. STUDY DESIGN/METHODS: Quantitative analysis of multi-components by a single-marker (QAMS) method was developed and validated for simultaneous determination of six flavonoids including narirutin, hesperidin, didymin, nobiletin, 3,5,6,7,8,3',4'-heptamethoxyflavone and tangeretin. Anti-lipase activity evaluation and docking studies of the flavonoids was also carried out to screen out the candidate lipase inhibitors. RESULTS: The QAMS method validation results exhibited that the developed method had desirable specificity, linearity, precision and accuracy. CRP collected in early months contained higher concentrations of bioactive flavonoids, and exhibited more potent anti-lipase activity. CONCLUSION: Harvest timing had a significant impact on the amounts of bioactive flavonoids and the anti-lipase activities of CRP extracts. The contents of total flavonoids were positively correlated with the anti-lipase activities of CRP, and polymethoxyflavones played a significant role in the hypolipidemic effect of CRP. Nobiletin might be the most potential lipase inhibitor in CRP.
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Citrus/química , Inhibidores Enzimáticos/farmacología , Flavonoides/farmacología , Lipasa/antagonistas & inhibidores , Agricultura , Animales , Biomarcadores Farmacológicos/análisis , Cromatografía Líquida de Alta Presión/métodos , Citrus/crecimiento & desarrollo , Evaluación Preclínica de Medicamentos/métodos , Medicamentos Herbarios Chinos/química , Medicamentos Herbarios Chinos/farmacología , Flavonoides/química , Frutas/química , Frutas/crecimiento & desarrollo , Reproducibilidad de los ResultadosRESUMEN
Detection of metabolites in complex biological matrixes is a great challenge because of the background noise and endogenous components. Herein, we proposed an integrated strategy that combined background subtraction program and modified mass defect filter (MMDF) data mining in a Microsoft Excel platform for chemicalome and metabolome profiling of the polymethoxylated flavonoids (PMFs) in Citri Reticulatae Pericarpium (CRP). The exogenously-sourced ions were firstly filtered out by the developed Visual Basic for Applications (VBA) program incorporated in the Microsoft Office. The novel MMDF strategy was proposed for detecting both target and untarget constituents and metabolites based on narrow, well-defined mass defect ranges. The approach was validated to be powerful, and potentially useful for the metabolite identification of both single compound and homologous compound mixture. We successfully identified 30 and 31 metabolites from rat biosamples after oral administration of nobiletin and tangeretin, respectively. A total of 56 PMFs compounds were chemically characterized and 125 metabolites were captured. This work demonstrated the feasibility of the integrated approach for reliable characterization of the constituents and metabolites in herbal medicines.
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Cromatografía Líquida de Alta Presión/métodos , Citrus/química , Medicamentos Herbarios Chinos/química , Flavonoides/química , Espectrometría de Masas/métodos , Metabolómica/métodos , Animales , Minería de Datos , Medicamentos Herbarios Chinos/metabolismo , Flavonas/análisis , Flavonas/metabolismo , Flavonoides/metabolismo , Masculino , Metaboloma , Peso Molecular , Ratas , Ratas Sprague-DawleyRESUMEN
To get a better understanding of the bioactive constituents in Aurantii Fructus Immaturus (AFI) and Aurantii Fructus (AF), in the present study, a comprehensive strategy integrating multiple chromatographic analysis and chemometrics methods was firstly proposed. Based on segmental monitoring, a high-performance liquid chromatography (HPLC)-variable wavelength detection method was established for simultaneous quantification of ten major flavonoids, and the quantitative data were further analyzed by hierarchical cluster analysis (HCA) and principal component analysis (PCA). A strong cation exchange-high performance liquid chromatography (SCX-HPLC) method combined with t-test and one-way analysis of variance (ANOVA) was developed to determine synephrine, the major alkaloid in AFI and AF. The essential oils were analyzed by gas chromatography-mass spectrometry (GC-MS) and further processed by partial least squares discrimination analysis (PLS-DA). The results indicated that the contents of ten flavonoids and synephrine in AFI were significantly higher than those in AF, and significant difference existed in samples from different geographical origins. Also, 9 differential volatile constituents detected could be used as chemical markers for discrimination of AFI and AF. Collectively, the proposed comprehensive analysis might be a well-acceptable strategy to evaluate the quality of traditional citrus herbs.
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Citrus/química , Flavonoides/análisis , Frutas/química , Sinefrina/análisis , Cromatografía Líquida de Alta Presión , Análisis por Conglomerados , Cromatografía de Gases y Espectrometría de Masas , Aceites Volátiles/química , Aceites de Plantas/química , Análisis de Componente PrincipalRESUMEN
Quassinoids, the predominant constituents in the seeds of Brucea javanica (BJ), have gained an increasing interest over the past decades since the discovery of their extensive biological activities. In the present study, a method based on the segment and exposure strategy coupled with two mass spectrometer data acquisition techniques was firstly developed and validated for comprehensive profiling of quassinoids in the seeds of BJ via high-performance liquid chromatography tandem quadrupole time-of-flight mass spectrometry (HPLC-QTOF/MS). The segment and exposure strategy could significantly improve the detection efficiency for trace quassinoids in the seeds of BJ. Furthermore, the five-point screening approach based on modified mass defect filter (MDF) and the visual isotopic ion technique could rapidly screen the precursor ions of interest. The fragmentation behavior of quassinoids was systematically investigated, and a total of 148 quassinoids including 86 potentially new ones were unambiguously or tentatively identified in the seeds of BJ. Collectively, our results demonstrate that the integrated strategy reported in this study has considerable potential for rapid screening of natural compounds especially for the trace ones in herbal medicines.
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Brucea/química , Cromatografía Líquida de Alta Presión/métodos , Espectrometría de Masas/métodos , Extractos Vegetales/química , Cuassinas/química , Semillas/química , Espectrometría de Masas/instrumentación , Estructura Molecular , Plantas Medicinales/químicaRESUMEN
Steroidal saponins, which exhibit multiple pharmacological effects, are the major bioactive constituents in herbal medicines from Dioscoreae species. In this study, a sensitive method based on high performance liquid chromatography-mass spectrometry (HPLC-MS) was established and validated for qualitative and quantitative analysis of steroidal saponins in four Dioscoreae herbs including Dioscoreae Nipponica Rhizome (DNR) and Dioscoreae Hypoglaucae Rhizome (DHR), Dioscoreae Spongiosae Rhizome (DSR) and Dioscoreae Rhizome (DR). A total of eleven steroidal saponins were identified by high performance liquid chromatography coupled with quadrupole time-of-flight mass spectrometry (HPLC-QTOF/MS). Furthermore, seven major steroidal saponins was simultaneous quantified using a high performance liquid chromatography coupled with triple quadrupole mass spectrometry (HPLC-QQQ/MS). The qualitative and quantitative analysis results indicated that the chemical composition of DNR, DHR and DSR samples exhibited a high level of global similarity, while the ingredients in DR varied greatly from the other three herbs. Moreover, principal component analysis (PCA) and hierarchical clustering analysis (HCA) were performed to compare and discriminate the Dioscoreae herbs based on the quantitative data. The results demonstrated the qualitative and quantitative analysis of steroidal saponins based on HPLC-MS is a feasible method for quality control of Dioscoreae herbs.
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Dioscoreaceae , Fitosteroles/análisis , Preparaciones de Plantas/análisis , Saponinas/análisis , Espectrometría de Masas en Tándem/métodos , Cromatografía Líquida de Alta Presión/métodosRESUMEN
The present study was designed to characterize the chemical constituents of Guge Fengtong Tablet (GGFTT). Based on the chromatographic retention behavior, fragmentation pathways of chemical components and the published literatures, a diagnostic ion filtering strategy with electrospray ionization quadrupole time-of-flight tandem mass spectrometry (HPLC-ESI-Q-TOF/MS) was established to identify the multiple bioactive constituents of GGFTT. The rapid identification of forty-seven components, including 18 phenolic acids, 8 saponins, 14 gingerol-related compounds, and 7 diarylhepatonoids, was accomplished using this newly developed method. The coupling of HPLC-ESI-Q-TOF/MS with the diagnostic ion filtering strategy was useful and efficient for the in-depth structural elucidation of chemical compounds of GGFTT.
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Medicamentos Herbarios Chinos/química , Catecoles/aislamiento & purificación , Cromatografía Líquida de Alta Presión , Diarilheptanoides/aislamiento & purificación , Alcoholes Grasos/aislamiento & purificación , Hidroxibenzoatos/aislamiento & purificación , Saponinas/aislamiento & purificación , Espectrometría de Masa por Ionización de Electrospray , Comprimidos Recubiertos/química , Espectrometría de Masas en TándemRESUMEN
The aim of this study was to develop a convenient method without pretreatments for nontarget discovery of interested compounds. The segment and exposure strategy, coupled with two mass spectrometer data acquisition methods was firstly proposed for screening the saponins in extract of Panax notoginseng (Sanqi) via high-performance liquid chromatography tandem quadrupole time-of-flight mass spectrometry (HPLC-QTOF/MS). By gradually removing certain major or moderate interference compounds, the developed segment and exposure strategy could significantly improve the detection efficiency for trace compounds. Moreover, the newly developed five-point screening approach based on a modified mass defect filter strategy and the visual isotopic ion technique was verified to be efficient and reliable in picking out the interested precursor ions. In total, 234 ginsenosides including 67 potential new ones were characterized or tentatively identified from the extract of Sanqi. Particularly, some unusual compounds containing the branched glycosyl group or new substituted acyl groups were firstly reported. The proposed integrated strategy held a strong promise for analyses of the complex mixtures.
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Medicamentos Herbarios Chinos/química , Panax notoginseng/química , Saponinas/análisis , Cromatografía Líquida de Alta Presión , Ginsenósidos/análisis , Indicadores y Reactivos , Espectrometría de Masas , Reproducibilidad de los ResultadosRESUMEN
In the present study, we developed and validated a rapid analytical method using high performance liquid chromatography coupled with quadrupole time of flight mass spectrometry (HPLC-Q-TOF/MS) to investigate the metabolic profile of Guge Fengtong tablet (GGFTT), a traditional Chinese medicine. The urine and bile samples were collected with 24 h after oral administration of GGFTT. A total of 34 compounds, including 11 parent compounds and 23 metabolites were unambiguously or tentatively identified. Our results indicated that glucuronidation, oxidation and methylation were the major metabolic pathways of the constituents in GGFTT. In addition, the results of this work also demonstrated the feasibility of HPLC-ESI-Q-TOF/MS for reliable characterization of the in vivo metabolites from herbal preparations.
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Bilis/química , Cromatografía Líquida de Alta Presión/métodos , Medicamentos Herbarios Chinos/química , Medicamentos Herbarios Chinos/metabolismo , Espectrometría de Masas en Tándem/métodos , Orina/química , Administración Oral , Animales , Bilis/metabolismo , Medicamentos Herbarios Chinos/administración & dosificación , Masculino , Metaboloma , Estructura Molecular , Ratas , Ratas Sprague-Dawley , Espectrometría de Masa por Ionización de Electrospray/métodos , Comprimidos/administración & dosificación , Comprimidos/química , Comprimidos/metabolismoRESUMEN
Relative quantification of multi-components in complex mixture is significantly affected by the ionization variance caused by mobile phase composition in high-performance liquid chromatography with electrospray ionization mass spectrometry (HPLC-ESI-MS) analyses. The normalization methods for eliminating the variance are still less investigated. Herein, the mobile-phase compensation (MPC) method was applied to overcome the above problem. The developed method was firstly used for convenient evaluation of the coeluent interference and subsequently applied for relative quantification of the identified multi-components in Panax notoginseng (Sanqi) samples. The good linearity, precision and low limit of quantification of targeted analytes confirmed that the MPC-HPLC-ESI-MS method in gradient elution could achieve the isocratic test results compared with classical HPLC-ESI-MS. The established method was used for relative quantification of the minor Sanqi saponins by their detected peak areas divided by that of ginsenoside Rd. The results demonstrated the potential of the newly developed method for obtaining the normalized data shared in different laboratories.
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Ginsenósidos/análisis , Panax notoginseng/química , Saponinas/análisis , Cromatografía Líquida de Alta Presión , Solventes/análisis , Espectrometría de Masa por Ionización de ElectrosprayRESUMEN
Absolute quantification of triacylglycerols (TAGs) in plant oils is a challenge for analysts, because most of the necessary chemical standards are unavailable. In this study, a new method for absolute quantification analysis of multi-components by use of a single marker (AQAMS), using two crucial technologies, evaluation of the collection recovery without chemical standards and enzymatic hydrolysis, was used for determining the absolute content of TAGs in brucea javanica oil (BJO), using glycerol as the marker. The TAGs in BJO were initially characterized using ultrafast liquid chromatography tandem atmospheric-pressure-chemical-ionization mass spectrometry. Then the TAGs in BJO were individually collected, by target-fraction collection via high-performance liquid chromatography coupled with an evaporative-light-scattering detector (HPLC-ELSD), and their recoveries were calculated by use of a novel non-standard evaluated recovery strategy (NSER). The results revealed that the collection procedure was feasible and reliable. Finally, modified commercial TAG assay kits using glycerol as the marker were used to determine the absolute abundance of individual TAGs in the plant oils. Comparing the result with that obtained by HPLC-ELSD analysis using triolein standard, the content of triolein determined by AQAMS was closely matched. The proposed strategy is a practical measure for solving the problem of the lack of chemical standards, and provides a new method for absolute quantification in natural products of multi-components with the same backbone.