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1.
J Agric Food Chem ; 67(46): 12972-12985, 2019 Nov 20.
Artigo em Inglês | MEDLINE | ID: mdl-31709797

RESUMO

Controlling the temperature inside a greenhouse during the summer is a problem of increasing importance in the Mediterranean countries, especially in the Spanish southeast. The metabolic profile of greenhouse tomatoes and leaves grown under conventional conditions and within the presence of a shade mesh (∼50% reduction of sunlight radiation) has been monitored. Tomatoes were weekly harvested from May to July 2017 and analyzed by NMR spectroscopy coupled to multivariate data analysis techniques, together with oxygen radical absorbance capacity (ORAC) assays (for antioxidant activity). Fatty acids and carotenoids profiles were unraveled by GC-FID and HPLC-DAD, respectively. To verify whether it would be possible to take advantage of different light growing conditions to potentiate a plant's defense system, leaves of the corresponding plants were collected and their methanolic extracts were analyzed by NMR toward deciphering new biomarkers, which were used to assess their antibacterial and antibiofilm activities. The presence of a shading mesh resulted in a reduction in tomato production and in smaller fruits with lower contents of sugars (glucose and fructose) and carotenoids (lycopene and ß-carotene) and higher contents of organic acids, amino acids, and polyunsaturated fatty acids (linoleic and oleic acids) and of phenylpropanoids and flavonoids (which contributed to an increased antioxidant activity). Methanolic extracts of leaves of nonshaded plants showed a higher antibiofilm activity than that from shaded plants. This activity was well-correlated with an increase of phenolic compounds, together with some specific amino acids and organic acids from tomato leaves.


Assuntos
Frutas/química , Lycopersicon esculentum/efeitos da radiação , Folhas de Planta/química , Antioxidantes/metabolismo , Alimentos Orgânicos/análise , Alimentos Orgânicos/efeitos da radiação , Frutas/crescimento & desenvolvimento , Frutas/metabolismo , Frutas/efeitos da radiação , Lycopersicon esculentum/química , Lycopersicon esculentum/crescimento & desenvolvimento , Lycopersicon esculentum/metabolismo , Espectroscopia de Ressonância Magnética , Metaboloma/efeitos da radiação , Metabolômica , Valor Nutritivo/efeitos da radiação , Extratos Vegetais/química , Folhas de Planta/crescimento & desenvolvimento , Folhas de Planta/metabolismo , Folhas de Planta/efeitos da radiação , Luz Solar
2.
Zhongguo Yi Xue Ke Xue Yuan Xue Bao ; 41(5): 709-713, 2019 Oct 30.
Artigo em Chinês | MEDLINE | ID: mdl-31699205

RESUMO

Renal cell carcinoma(RCC)is one of the most common malignant tumors in the urinary system.Many patients have already been in the advanced stage at the first medical consultation and the prognosis is dismal.Metabonomics searches for differential metabolomes through high-throughput analysis of endogenous metabolites showing high potential in the early diagnosis of RCC and the investigations on its pathophysiological mechanisms.Metabonomics techniques are also useful for identifying tumor markers,which will help to enable early diagnosis and improve clinical prognosis of RCC.


Assuntos
Carcinoma de Células Renais/diagnóstico , Neoplasias Renais/diagnóstico , Metaboloma , Biomarcadores Tumorais , Detecção Precoce de Câncer , Humanos , Prognóstico
3.
J Agric Food Chem ; 67(44): 12199-12207, 2019 Nov 06.
Artigo em Inglês | MEDLINE | ID: mdl-31595753

RESUMO

Salvianolic acid A (Sal A) has a wide range of pharmacological activities. To date, there have been no systematic and detailed metabolite research data of Sal A after oral administration in vitro and in vivo. In this study, a rapid and systematic method based on ultrafast liquid chromatography-quadrupole-time-of-flight mass spectrometry was developed to detect metabolites of Sal A in vitro (human liver microsome, human intestinal microbiota, artificial gastric, and intestinal juice) and in vivo (urine, plasma, feces, and various organs collected after oral administration of Sal A to normal rats and pseudo-germ-free rats). A total of 26 metabolites of Sal A were characterized. These metabolites were formed through extensive metabolic reactions, such as hydroxylation, hydrogenation, and glucuronidation reactions. This study provides novel possibility for exploring the potential biological mechanism of Sal A, and aids the promotion of clinical application.


Assuntos
Ácidos Cafeicos/química , Ácidos Cafeicos/metabolismo , Cromatografia Líquida de Alta Pressão/métodos , Medicamentos de Ervas Chinesas/química , Medicamentos de Ervas Chinesas/metabolismo , Lactatos/química , Lactatos/metabolismo , Espectrometria de Massas/métodos , Salvia miltiorrhiza/química , Adulto , Animais , Feminino , Humanos , Masculino , Metaboloma , Microssomos Hepáticos/química , Microssomos Hepáticos/metabolismo , Ratos , Ratos Sprague-Dawley , Adulto Jovem
4.
Life Sci ; 237: 116914, 2019 Nov 15.
Artigo em Inglês | MEDLINE | ID: mdl-31622606

RESUMO

AIMS: The aim of the presente study was to examine the effects of oral gallic acid (GA) administration on the brown adipose tissue of obese mice fed with high-fat diet. New mechanisms and interactions pathways in thermogenesis were accessed through bioinformatics analyses. MAIN METHODS: Swiss male mice were divided into four groups and fed during 60 days with: standard diet, standard diet combined with gallic acid, high-fat diet and high-fat diet combined with gallic acid. Body weight, food intake, and blood parameters (glucose tolerance test, total-cholesterol, high-density low-c, triglyceride and glucose levels) were evaluated. Brown and subcutaneous white adipose tissue histological analysis were performed. SIRT1 and PGC1-α mRNA expression in the brown adipose tissue were assessed. KEY FINDINGS: Our main findings showed that the gallic acid improved glucose tolerance and metabolic parameters. These results were accompanied by bioinformatics analyses that evidenced SIRT1 as main target in the thermogenesis process, confirmed as increased SIRT1 mRNA expression was evidenced in the brown adipose tissue. SIGNIFICANCE: Together, the data suggest that the gallic acid effect in brown adipose tissue may improve body metabolism, glucose homeostasis and increase thermogenesis.


Assuntos
Tecido Adiposo Marrom/metabolismo , Biologia Computacional/métodos , Dieta Hiperlipídica/efeitos adversos , Ácido Gálico/farmacologia , Metaboloma/efeitos dos fármacos , Obesidade/metabolismo , Sirtuína 1/metabolismo , Tecido Adiposo Marrom/efeitos dos fármacos , Tecido Adiposo Marrom/patologia , Animais , Regulação da Expressão Gênica/efeitos dos fármacos , Masculino , Camundongos , Camundongos Obesos , Obesidade/tratamento farmacológico , Obesidade/etiologia , Sirtuína 1/genética , Termogênese/efeitos dos fármacos
5.
Life Sci ; 236: 116939, 2019 Nov 01.
Artigo em Inglês | MEDLINE | ID: mdl-31593705

RESUMO

Taxifolin (TAX) reportedly exerts protective and therapeutic effects in liver. Herein, the effects of TAX against acetaminophen (APAP)-induced hepatotoxicity were investigated. Pharmacodynamics, pharmacology and metabolomics analyses of TAX were assessed on C57 mice and L-02 cells. TAX was administered for 7 days, and APAP was given on the last day to establish an acute liver injury model. ALT and AST levels were determined, and liver ROS, MDA, GST, GSH and GPX1 were analysed. The expression and protein abundance of GPX1, GPS-Pi, GCLC and GCLM were assessed by PCR and western blotting, and metabolic changes in cells and serum were investigated by UPLC-Q-Orbitrap-MS. Serum ALT and AST, and liver ROS, MDA, GST, GSH and GPX1 levels confirmed the protective effects of TAX. Besides, we found Only treating with TAX decreased the expression of CYP2E1 in mice liver tissue. TAX reversed the APAP-induced decrease in cell viability in L-02 cells, and reduced cellular ROS levels. Furthermore, TAX reversed the APAP-induced decrease in antioxidant enzymes at both mRNA and protein levels. Metabolomics analysis identified metabolites mainly related to glutathione metabolism (36 in vivo and 23 in vitro). The concentration of glutathione, oxidized glutathione, carnitine, succinic acid, pyroglutamic acid, citrulline, taurine, palmitoleic acid, phytoshingosine-1-P and sphingosine-1-P were close to normal levels after treating with TAX. These results indicate that TAX prevents APAP-induced liver injury by inhibiting APAP metabolic activation mediated by CYP450 enzymes, modulating glutathione metabolism, and expression of related antioxidative signals. These properties could be harnessed to prevent or treat hepatotoxicity.


Assuntos
Acetaminofen/toxicidade , Doença Hepática Induzida por Substâncias e Drogas/prevenção & controle , Glutationa/metabolismo , Metaboloma , Substâncias Protetoras/farmacologia , Quercetina/análogos & derivados , Analgésicos não Entorpecentes/toxicidade , Animais , Anti-Inflamatórios não Esteroides/farmacologia , Doença Hepática Induzida por Substâncias e Drogas/etiologia , Doença Hepática Induzida por Substâncias e Drogas/metabolismo , Doença Hepática Induzida por Substâncias e Drogas/patologia , Camundongos , Camundongos Endogâmicos C57BL , Estresse Oxidativo/efeitos dos fármacos , Quercetina/farmacologia , Espécies Reativas de Oxigênio/metabolismo
6.
Nat Biotechnol ; 37(10): 1217-1228, 2019 10.
Artigo em Inglês | MEDLINE | ID: mdl-31477923

RESUMO

Depleted gut microbiome α-diversity is associated with several human diseases, but the extent to which this is reflected in the host molecular phenotype is poorly understood. We attempted to predict gut microbiome α-diversity from ~1,000 blood analytes (laboratory tests, proteomics and metabolomics) in a cohort enrolled in a consumer wellness program (N = 399). Although 77 standard clinical laboratory tests and 263 plasma proteins could not accurately predict gut α-diversity, we found that 45% of the variance in α-diversity was explained by a subset of 40 plasma metabolites (13 of the 40 of microbial origin). The prediction capacity of these 40 metabolites was confirmed in a separate validation cohort (N = 540) and across disease states, showing that our findings are robust. Several of the metabolite biomarkers that are reported here are linked with cardiovascular disease, diabetes and kidney function. Associations between host metabolites and gut microbiome α-diversity were modified in those with extreme obesity (body mass index ≥ 35), suggesting metabolic perturbation. The ability of the blood metabolome to predict gut microbiome α-diversity could pave the way to the development of clinical tests for monitoring gut microbial health.


Assuntos
Bactérias/classificação , Microbioma Gastrointestinal , Metaboloma , Bactérias/genética , Estudos de Coortes , Variação Genética , Humanos , Metabolômica , RNA Ribossômico 16S/sangue , RNA Ribossômico 16S/genética
7.
Nat Med ; 25(9): 1442-1452, 2019 09.
Artigo em Inglês | MEDLINE | ID: mdl-31477907

RESUMO

Our understanding of how the gut microbiome interacts with its human host has been restrained by limited access to longitudinal datasets to examine stability and dynamics, and by having only a few isolates to test mechanistic hypotheses. Here, we present the Broad Institute-OpenBiome Microbiome Library (BIO-ML), a comprehensive collection of 7,758 gut bacterial isolates paired with 3,632 genome sequences and longitudinal multi-omics data. We show that microbial species maintain stable population sizes within and across humans and that commonly used 'omics' survey methods are more reliable when using averages over multiple days of sampling. Variation of gut metabolites within people over time is associated with amino acid levels, and differences across people are associated with differences in bile acids. Finally, we show that genomic diversification can be used to infer eco-evolutionary dynamics and in vivo selection pressures for strains within individuals. The BIO-ML is a unique resource designed to enable hypothesis-driven microbiome research.


Assuntos
Bactérias/genética , Microbioma Gastrointestinal/genética , Filogenia , Seleção Genética/genética , Bactérias/classificação , Bactérias/isolamento & purificação , Ácidos e Sais Biliares/genética , Ácidos e Sais Biliares/metabolismo , Bancos de Espécimes Biológicos , Fezes/microbiologia , Variação Genética/genética , Genoma Bacteriano/genética , Humanos , Metaboloma/genética
8.
J Agric Food Chem ; 67(38): 10667-10677, 2019 Sep 25.
Artigo em Inglês | MEDLINE | ID: mdl-31483636

RESUMO

This study investigated the modulatory effects of Decaisnea insignis seed oil (DISO), which was rich in palmitoleic acid (55.25%), palmitic acid (12.25%), and oleic acid (28.74%), on alcohol-induced metabolism disorder in mice. Fifty mice were orally administered with 38% alcohol (0.4 mL/day) and without or with DISO (3, 6, and 12 g/kg) for consecutive 12 weeks. DISO inhibited the alcohol-induced weight loss and liver function abnormality (p < 0.01) and shifted the profiles of cecal microbiome: elevating the abundance of Lactobacillus, Ruminoccoceae_UCG_004 (p < 0.05) and decreasing abundance of Parabacteroides (p < 0.05). This treatment also regulated metabolome response of amino acid and lipid metabolism in cecal content: upregulating 5-hydroxyindole-3-acetic acid (p < 0.05), 6-hydroxynicotinic acid, 5-methoxytryptamine, nicotinamide, and nicotinic acid (p < 0.1) and downregulating androsterone, tryptophan, and indole-3-acetamide (p < 0.05). DISO protected against alcoholic liver injury and gut microbiota dysbiosis by enriching the relative abundance of Lactobacillus, which was positively associated with the improvement of intestinal permeability and tryptophan metabolism.


Assuntos
Álcoois/efeitos adversos , Disbiose/prevenção & controle , Microbioma Gastrointestinal/efeitos dos fármacos , Hepatopatias Alcoólicas/prevenção & controle , Magnoliopsida/química , Óleos Vegetais/administração & dosagem , Consumo de Bebidas Alcoólicas/efeitos adversos , Aminoácidos/metabolismo , Animais , Bactérias/classificação , Bactérias/efeitos dos fármacos , Bactérias/genética , Bactérias/isolamento & purificação , Ceco/efeitos dos fármacos , Ceco/microbiologia , Disbiose/metabolismo , Disbiose/microbiologia , Humanos , Metabolismo dos Lipídeos/efeitos dos fármacos , Fígado/efeitos dos fármacos , Fígado/metabolismo , Hepatopatias Alcoólicas/metabolismo , Hepatopatias Alcoólicas/microbiologia , Masculino , Metaboloma/efeitos dos fármacos , Camundongos , Microbiota/efeitos dos fármacos , Sementes/química
9.
Pharm Res ; 36(11): 158, 2019 Sep 11.
Artigo em Inglês | MEDLINE | ID: mdl-31512001

RESUMO

PURPOSE: P-glycoprotein (P-gp) and breast cancer resistance protein (BCRP) are xenobiotic transporters which pump out variety types of compounds, but information on their interaction with endogenous substrates in the skin is limited. The purpose of the present study was to clarify possible association of these transporters in dermal accumulation of inflammatory mediators. METHODS: Dermatitis model was constructed by repeated topical application of oxazolone in wild-type, and P-gp and BCRP gene triple knockout (Mdr1a/1b/Bcrp-/-) mice to observe difference in phenotype. Target metabolome analysis of 583 metabolites was performed using skin and plasma. RESULTS: Dermatitis and scratching behavior in dermatitis model of Mdr1a/1b/Bcrp-/- mice were more severe than wild-type mice, suggesting protective roles of these transporters. This hypothesis was supported by the metabolome analysis which revealed that concentration of histamine and other dermatitis-associated metabolites like urate and serotonin in the dermatitis skin, but not normal skin, of Mdr1a/1b/Bcrp-/- mice was higher than that of wild-type mice. Gene expression of P-gp and BCRP was reduced in oxazolone-treated skin and the skin of patients with atopic dermatitis or psoriasis. CONCLUSIONS: These results suggest possible association of these efflux transporters with dermal inflammatory mediators, and such association could be observed in the dermatitis skin.


Assuntos
Membro 1 da Subfamília B de Cassetes de Ligação de ATP/genética , Membro 2 da Subfamília G de Transportadores de Cassetes de Ligação de ATP/genética , Dermatite/metabolismo , Histamina/metabolismo , Metaboloma/efeitos dos fármacos , Proteínas de Neoplasias/genética , Pele/metabolismo , Animais , Deleção de Genes , Humanos , Masculino , Camundongos , Camundongos Knockout
10.
Int J Nanomedicine ; 14: 6297-6311, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-31496687

RESUMO

Purpose: Mn-doped ZnS quantum dots (QDs) with special luminescent properties have been widely researched and applied in various fields. Thus, their release toxicity and security cannot be ignored. Methods: In the present study, the toxicity and non-targeted metabolomics of Mn-doped ZnS QDs were investigated after single intravenous injection. Serum metabolites were evaluated based on gas chromatography-mass spectrometry together with multivariate statistical analyses [principal component analysis, partial least squares discriminant analysis, and orthogonal PLS-DA]. Results: The modified metabolites (variable importance in the projection (VIP) >1 and p<0.05) revealed that Mn-doped ZnS QDs exposure disturbed glycolysis, tricarboxylic acid cycle, ketoplasia, glutaminolysis, and amino acid and lipid metabolism. The behavior, coefficients of organs, and histological changes were the same as in the control group, and the disturbance of hematology and serum biochemistry was not dose- or time-dependent. Conclusion: Our study provides a general observation regarding the toxicity and potential metabolic responses of mice exposed to Mn-doped ZnS QDs.


Assuntos
Manganês/química , Metabolômica , Pontos Quânticos/toxicidade , Sulfetos/sangue , Sulfetos/toxicidade , Compostos de Zinco/sangue , Compostos de Zinco/toxicidade , Animais , Peso Corporal/efeitos dos fármacos , Masculino , Metaboloma , Camundongos Endogâmicos BALB C , Análise Multivariada , Tamanho do Órgão/efeitos dos fármacos , Pontos Quânticos/administração & dosagem , Pontos Quânticos/química , Sulfetos/administração & dosagem , Compostos de Zinco/administração & dosagem
11.
J Agric Food Chem ; 67(41): 11542-11552, 2019 Oct 16.
Artigo em Inglês | MEDLINE | ID: mdl-31538781

RESUMO

Two field trials were conducted to investigate the influence of fungicide and fertilization management on the potato tubers' metabolome (Solanum tuberosum L.). Thereby, fungicides and conventional fertilizers were varied in terms of quantities, number and date of applications, physical state, and product composition. Following a water-methanol-based extraction, samples were analyzed using an UPLC-IMS-QToF and multivariate data analysis. Fungicide application led to significant changes in the tubers' metabolome. Flavonoids were increasingly expressed as a natural response to impending fungal or viral infections in an untreated group, while the phytoalexin rishitinol was highly abundant in groups with fungicide application. In contrast to fungicides, the application of conventional fertilizers did not cause significant alterations in the tubers' compound composition. Consequently, the impact of fungicide application could be rated as more important than the fertilization-derived influence, which might be because of a gentler adaption to fertilization than to the acute stress of fungicide applications.


Assuntos
Produção Agrícola/métodos , Fertilizantes/análise , Fungicidas Industriais/farmacologia , Espectrometria de Massas/métodos , Tubérculos/química , Solanum tuberosum/efeitos dos fármacos , Cromatografia Líquida de Alta Pressão , Metaboloma , Tubérculos/efeitos dos fármacos , Tubérculos/crescimento & desenvolvimento , Tubérculos/metabolismo , Solanum tuberosum/química , Solanum tuberosum/crescimento & desenvolvimento , Solanum tuberosum/metabolismo
12.
Nat Methods ; 16(8): 743-749, 2019 08.
Artigo em Inglês | MEDLINE | ID: mdl-31363225

RESUMO

Cellular behavior is controlled by the interplay of diverse biomolecules. Most experimental methods, however, can only monitor a single molecule class or reaction type at a time. We developed an in vitro nuclear magnetic resonance spectroscopy (NMR) approach, which permitted dynamic quantification of an entire 'heterotypic' network-simultaneously monitoring three distinct molecule classes (metabolites, proteins and RNA) and all elementary reaction types (bimolecular interactions, catalysis, unimolecular changes). Focusing on an eight-reaction co-transcriptional RNA folding network, in a single sample we recorded over 35 time points with over 170 observables each, and accurately determined five core reaction constants in multiplex. This reconstruction revealed unexpected cross-talk between the different reactions. We further observed dynamic phase-separation in a system of five distinct RNA-binding domains in the course of the RNA transcription reaction. Our Systems NMR approach provides a deeper understanding of biological network dynamics by combining the dynamic resolution of biochemical assays and the multiplexing ability of 'omics'.


Assuntos
Redes Reguladoras de Genes , Metaboloma , Ressonância Magnética Nuclear Biomolecular/métodos , Proteínas/análise , RNA/análise , Células HEK293 , Humanos , Conformação de Ácido Nucleico , Conformação Proteica , Proteínas/química , RNA/química , Dobramento de RNA
13.
Adv Exp Med Biol ; 1155: 523-529, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-31468428

RESUMO

Mammalian tissues, especially the heart, contain high concentrations of taurine, a beta-amino acid that possesses a variety of physiological functions. While it is well known that taurine reacts with several metabolites, such as bile acids and fatty acids, taurine-conjugated metabolites in the heart have not been specifically studied. Recently, we performed Liquid chromatography-mass spectrometry- (LC-MS-) based metabolome analysis, comparing metabolome profiles of hearts from taurine transporter knockout (TauTKO) mice and wild-type mice to identify differences in taurine-conjugated metabolite content of the two phenotypes. Comparison of the metabolite profiles revealed taurine-containing dipeptides, such as glutamyltaurine, which are present in wild-type but not in TauTKO hearts. These data suggest that taurine functions not only as a free osmolyte but also as a conjugated metabolite within the heart.


Assuntos
Coração , Metaboloma , Miocárdio/metabolismo , Taurina/metabolismo , Animais , Cromatografia Líquida , Camundongos , Camundongos Knockout , Espectrometria de Massas em Tandem
14.
BMC Plant Biol ; 19(1): 343, 2019 Aug 06.
Artigo em Inglês | MEDLINE | ID: mdl-31387524

RESUMO

BACKGROUND: European grapevine cultivars (Vitis vinifera spp.) are highly susceptible to the downy mildew pathogen Plasmopara viticola. Breeding of resistant V. vinifera cultivars is a promising strategy to reduce the impact of disease management. Most cultivars that have been bred for resistance to downy mildew, rely on resistance mediated by the Rpv3 (Resistance to P. viticola) locus. However, despite the extensive use of this locus, little is known about the mechanism of Rpv3-mediated resistance. RESULTS: In this study, Rpv3-mediated defense responses were investigated in Rpv3+ and Rpv3- grapevine cultivars following inoculation with two distinct P. viticola isolates avrRpv3+ and avrRpv3-, with the latter being able to overcome Rpv3 resistance. Based on comparative microscopic, metabolomic and transcriptomic analyses, our results show that the Rpv3-1-mediated resistance is associated with a defense mechanism that triggers synthesis of fungi-toxic stilbenes and programmed cell death (PCD), resulting in reduced but not suppressed pathogen growth and development. Functional annotation of the encoded protein sequence of genes significantly upregulated during the Rpv3-1-mediated defense response revealed putative roles in pathogen recognition, signal transduction and defense responses. CONCLUSION: This study used histochemical, transcriptomic and metabolomic analyses of Rpv3+ and susceptible cultivars inoculated with avirulent and virulent P. viticola isolates to investigate mechanism underlying the Rpv3-1-mediated resistance response. We demonstrated a strong correlation between the expressions of stilbene biosynthesis related genes, the accumulation of fungi-toxic stilbenes, pathogen growth inhibition and PCD.


Assuntos
Resistência à Doença/genética , Genes de Plantas/fisiologia , Estilbenos/metabolismo , Vitis/genética , Regulação da Expressão Gênica de Plantas , Metaboloma , Oomicetos/patogenicidade , Doenças das Plantas/microbiologia , Transcrição Genética , Transcriptoma , Vitis/imunologia , Vitis/microbiologia
15.
Life Sci ; 234: 116751, 2019 Oct 01.
Artigo em Inglês | MEDLINE | ID: mdl-31415771

RESUMO

AIMS: The present study aims to investigate the impacts of olfactory bulbectomy (OBX) on urinary metabolic profile and tryptophan metabolites in prefrontal cortex (PFC) of rats, and to explore the regulation effects of fluoxetine. MAIN METHODS: OBX model was developed by aspiration of olfactory bulbs. After fluoxetine treatment (10 mg/kg) for 14 days, urine samples were collected and behavior tests were applied. Tryptophan (TRP) metabolites and neurotransmitters in PFC were determined by prominence ultrafast liquid chromatography-QTRAP-mass spectrometry, and tryptophan hydroxylase 2 (TPH2) and indoleamine-2,3-dioxygenase 1 (IDO1) were evaluated by western blot. Urinary metabolites were analyzed by ultra-performance liquid chromatography coupled with quadrupole time-of-flight mass spectrometry-based metabonomics strategy. KEY FINDING: OBX rats showed hyperlocomotion in open field, hyperactivity in open arm and despair status, and fluoxetine reserved these behavioral abnormalities. The levels of TRP, 5-HIAA, 5-HIAA/5-HT ratio and DA increased, while kynurenine and 5-HT decreased in PFC of OBX rats. The activities of TPH2 and IDO1were inhibited after OBX. Twenty-six altered metabolites were identified as potential biomarkers in OBX rats involved in tryptophan metabolism, gut microbiota metabolism, energy metabolism, purine metabolism, ascorbate and aldarate metabolism, and tyrosine metabolism. Among them, 15 abnormal metabolites were corrected by fluoxetine to some extent. SIGNIFICANCE: Our results revealed that urinary metabolic profile changed greatly in OBX rats, and identified biomarkers might be helpful for the diagnosis of agitated depression. The regulation effects of fluoxetine on urinary metabolic profile and tryptophan metabolites in PFC might contribute to its antidepressant action in OBX rats.


Assuntos
Antidepressivos de Segunda Geração/uso terapêutico , Depressão/tratamento farmacológico , Depressão/metabolismo , Fluoxetina/uso terapêutico , Metaboloma/efeitos dos fármacos , Animais , Antidepressivos de Segunda Geração/farmacologia , Depressão/urina , Modelos Animais de Doenças , Fluoxetina/farmacologia , Masculino , Bulbo Olfatório/cirurgia , Córtex Pré-Frontal/efeitos dos fármacos , Córtex Pré-Frontal/metabolismo , Ratos Sprague-Dawley , Triptofano/metabolismo
16.
Anal Bioanal Chem ; 411(23): 6189-6202, 2019 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-31414155

RESUMO

It is a challenge to expand the metabolome coverage of liquid chromatography (LC)-electrospray ionization (ESI) mass spectrometry (MS) based untargeted metabolomics analysis. The limited coverage is attributed to the weak signal of hydroxyl and carboxyl groups in ESI-MS and the limited capacity of LC separation for metabolites with a wide range of polarities. Here a new sample preparation procedure is proposed to solve these problems. Mixed-mode (reversed-phase and anion-exchange) solid-phase extraction sorbents were used to separate metabolites into hydrophilic amine, hydrophobic amine/alcohol, and organic acid groups. Then, alcohols and carboxylic acids in separated groups were tagged with pyridine with use of two derivatization systems for signal enhancement. Finally, hydrophilic amines were analyzed by LC-MS with a hydrophilic interaction LC column, and the two hydrophobic compound groups were analyzed by LC-MS with a C18 column. From the results for standard samples, the detection limits of the new method are lower than those of the classic solvent extraction-protein precipitation method by 3.3-70 times for five amino acids and by 65-1141 times for five fatty acids. Moreover, the detection limit of this new method is 125 ng mL-1 for cholesterol, which has no signal with the classic method even at 10 µg mL-1. In seminal plasma samples, 110 more metabolites were identified by this new method than by the traditional solvent extraction-protein precipitation method in positive-mode ESI (new method vs traditional method, 65 vs 22 identified by comparing MS/MS spectra with those of standards, 203 vs 136 identified by searching MS spectra in a published database). Among them, 53 carboxylic acids and 21 alcohols were identified only by the new method, and more hydrophilic amine metabolites, such as amino acids and nucleosides, were identified by the new method than by the classic method. Finally, in application to the study of male infertility, more potential biomarkers of oligoasthenoteratospermic infertility were found with the new method (46 potential biomarkers) than with the classic method (19 potential biomarkers) and previously reported methods (10-30 potential biomarkers). Thus, it is demonstrated that this new sample preparation method expands the detection coverage of LC-MS-based untargeted metabolomics methods and has application potential in biological research.


Assuntos
Infertilidade Masculina/metabolismo , Metaboloma , Metabolômica/métodos , Sêmen/metabolismo , Biomarcadores/metabolismo , Humanos , Masculino , Sêmen/química , Extração em Fase Sólida/métodos , Espectrometria de Massas por Ionização por Electrospray/métodos
17.
Physiol Rev ; 99(4): 1819-1875, 2019 10 01.
Artigo em Inglês | MEDLINE | ID: mdl-31434538

RESUMO

Metabolomics uses advanced analytical chemistry techniques to enable the high-throughput characterization of metabolites from cells, organs, tissues, or biofluids. The rapid growth in metabolomics is leading to a renewed interest in metabolism and the role that small molecule metabolites play in many biological processes. As a result, traditional views of metabolites as being simply the "bricks and mortar" of cells or just the fuel for cellular energetics are being upended. Indeed, metabolites appear to have much more varied and far more important roles as signaling molecules, immune modulators, endogenous toxins, and environmental sensors. This review explores how metabolomics is yielding important new insights into a number of important biological and physiological processes. In particular, a major focus is on illustrating how metabolomics and discoveries made through metabolomics are improving our understanding of both normal physiology and the pathophysiology of many diseases. These discoveries are yielding new insights into how metabolites influence organ function, immune function, nutrient sensing, and gut physiology. Collectively, this work is leading to a much more unified and system-wide perspective of biology wherein metabolites, proteins, and genes are understood to interact synergistically to modify the actions and functions of organelles, organs, and organisms.


Assuntos
Metabolismo Energético , Metaboloma , Metabolômica/métodos , Animais , Biomarcadores/metabolismo , Doenças Cardiovasculares/metabolismo , Doenças Cardiovasculares/fisiopatologia , Humanos , Nefropatias/metabolismo , Nefropatias/fisiopatologia , Erros Inatos do Metabolismo/metabolismo , Erros Inatos do Metabolismo/fisiopatologia , Neoplasias/metabolismo , Neoplasias/fisiopatologia , Fluxo de Trabalho
18.
Anal Bioanal Chem ; 411(26): 6857-6866, 2019 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-31420709

RESUMO

We show an alternative way to visualize time course NMR data without the application of multivariate data analysis, based on the temporal change of the metabolome of hazelnuts after mold infestation. Fresh hazelnuts were inoculated with eight different natural mold species and the growth was studied over a period of 14 days. The data were plotted in a color-coded scheme showing metabolic changes as a function of chemical shift, which we named signal pattern plot. This plot graphically displays alteration (trend) of a respected signal over time and allows visual interpretation in a simple manner. Changes are compared with a reference sample stored under identical conditions as the infected nuts. The plot allows, at a glance, the recognition of individual landmarks specific to a sample group as well as common features of the spectra. Each sample reveals an individual signal pattern. The plot facilitates the recognition of signals that belong to biological relevant metabolites. Betaine and five signals were identified that specifically changed upon mold infestation. Graphical abstract.


Assuntos
Corylus/metabolismo , Corylus/microbiologia , Metaboloma , Metabolômica/métodos , Espectroscopia de Prótons por Ressonância Magnética/métodos , Aspergillus niger/fisiologia , Betaína/análise , Betaína/metabolismo , Corylus/química , Fungos/fisiologia , Interações Hospedeiro-Patógeno , Doenças das Plantas/microbiologia
19.
Anal Bioanal Chem ; 411(26): 6983-6994, 2019 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-31463516

RESUMO

This study investigated the optimal inter-batch normalization method for gas chromatography/tandem mass spectrometry (GC/MS/MS)-based targeted metabolome analysis of rodent blood samples. The effect of centrifugal concentration on inter-batch variation was also investigated. Six serum samples prepared from a mouse and 2 quality control (QC) samples from pooled mouse serum were assigned to each batch, and the 3 batches were analyzed by GC/MS/MS at different days. The following inter-batch normalization methods were applied to metabolome data: QC-based methods with quadratic (QUAD)- or cubic spline (CS)-fitting, total signal intensity (TI)-based method, median signal intensity (MI)-based method, and isotope labeled internal standard (IS)-based method. We revealed that centrifugal concentration was a critical factor to cause inter-batch variation. Unexpectedly, neither the QC-based normalization methods nor the IS-based method was able to normalize inter-batch variation, though MI- or TI-based normalization methods were effective in normalizing inter-batch variation. For further validation, 6 disease model rat and 6 control rat plasma were evenly divided into 3 batches, and analyzed as different batches. Same as the results above, MI- or TI-based methods were able to normalize inter-batch variation. In particular, the data normalized by TI-based method showed similar metabolic profiles obtained from their intra-batch analysis. In conclusion, the TI-based normalization method is the most effective to normalize inter-batch variation for GC/MS/MS-based metabolome analysis. Graphical abstract.


Assuntos
Metaboloma , Metabolômica/métodos , Plasma/metabolismo , Soro/metabolismo , Animais , Centrifugação/métodos , Cromatografia Gasosa-Espectrometria de Massas/métodos , Masculino , Camundongos Endogâmicos ICR , Controle de Qualidade , Ratos , Síndrome da Serotonina/sangue , Síndrome da Serotonina/metabolismo , Espectrometria de Massas em Tandem/métodos
20.
Ecotoxicol Environ Saf ; 183: 109501, 2019 Nov 15.
Artigo em Inglês | MEDLINE | ID: mdl-31401330

RESUMO

17α-ethynylestradiol (EE2), a ubiquitous synthetic endocrine disrupting chemical, was the principal component of contraceptive drugs and one of common hormone medications. The detrimental impact of EE2 on the reproduction of organisms was widely recognized. However, the underlying mechanisms of physiological and metabolome effects of EE2 on freshwater fish are still unclear. This study investigated the toxic effects and related mechanisms of EE2 on freshwater fish crucian carp (Carassius auratus) based on metabolomics. Crucian carp were exposed to EE2 at environmentally relevant concentration for 9 days, 18 days, and 27 days, and the biological responses were explored through analysis of the physiological endpoints, steroid hormones, and metabolome. The physiological endpoints of crucian carp had no distinct change after EE2 exposure. However, metabolomics analysis probed significant deviation based on chemometrics, indicating that the metabolomics approach was more sensitive to the effects of EE2 at environmentally relevant concentration to freshwater fish than the traditional endpoints. The alterations of 24 metabolites in gonad and 16 metabolites in kidney were induced by treatment with EE2, respectively, which suggesting the perturbations in amino acid metabolism, lipid metabolism, energy metabolism, and oxidative stress. Moreover, EE2 exposure could induce the disruption of lipid metabolism and then broke the homeostasis of endogenous steroid hormones. Metabolomics provided a new strategy for the studies on contaminant exposure at a low dose in the short term and gave important information for the toxicology and mechanism of EE2.


Assuntos
Disruptores Endócrinos/toxicidade , Etinilestradiol/toxicidade , Carpa Dourada/metabolismo , Metaboloma/efeitos dos fármacos , Poluentes Químicos da Água/toxicidade , Animais , Exposição Ambiental/efeitos adversos , Metabolômica , Reprodução/efeitos dos fármacos , Esteroides/metabolismo
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