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1.
Int J Mol Sci ; 22(15)2021 Jul 26.
Artigo em Inglês | MEDLINE | ID: mdl-34360755

RESUMO

Increasing attention is being focused on the use of polypeptide-based N-methyl-d-aspartate (NMDA) receptor antagonists for the treatment of nervous system disorders. In our study on Achyranthes bidentata Blume, we identified an NMDA receptor subtype 2B (NR2B) antagonist that exerts distinct neuroprotective actions. This antagonist is a 33 amino acid peptide, named bidentatide, which contains three disulfide bridges that form a cysteine knot motif. We determined the neuroactive potential of bidentatide by evaluating its in vitro effects against NMDA-mediated excitotoxicity. The results showed that pretreating primary cultured hippocampal neurons with bidentatide prevented NMDA-induced cell death and apoptosis via multiple mechanisms that involved intracellular Ca2+ inhibition, NMDA current inhibition, and apoptosis-related protein expression regulation. These mechanisms were all dependent on bidentatide-induced inhibitory regulation of NR2B-containing NMDA receptors; thus, bidentatide may contribute to the development of neuroprotective agents that would likely possess the high selectivity and safety profiles inherent in peptide drugs.


Assuntos
Achyranthes/química , Hipocampo/metabolismo , Neurônios/metabolismo , Fármacos Neuroprotetores , Peptídeos , Proteínas de Plantas , Receptores de N-Metil-D-Aspartato/antagonistas & inibidores , Animais , Apoptose/efeitos dos fármacos , Cálcio/metabolismo , Fármacos Neuroprotetores/química , Fármacos Neuroprotetores/isolamento & purificação , Fármacos Neuroprotetores/farmacologia , Peptídeos/química , Peptídeos/isolamento & purificação , Peptídeos/farmacologia , Proteínas de Plantas/química , Proteínas de Plantas/isolamento & purificação , Proteínas de Plantas/farmacologia , Ratos , Ratos Sprague-Dawley , Receptores de N-Metil-D-Aspartato/metabolismo
2.
Proteomics Clin Appl ; : e2000058, 2021 Jul 30.
Artigo em Inglês | MEDLINE | ID: mdl-34329527

RESUMO

PURPOSE: There are great demands for identifying biomarkers of major depressive disorder (MDD), a common mental illness with a prevalence of approximately 6%. Finding potential biomarkers to aid MDD diagnosis is in high demand. EXPERIMENTAL DESIGN: In this study, a combination of pretreatment methods named salt-out assisted liquid-liquid extraction (SALLE) and nontargeted peptidomics based on nano-LC-Orbitrap/MS was primarily employed to discover the candidate peptide markers from the plasma of 238 subjects. RESULTS: Many peptides were enriched and identified from the plasma, 42 of which showed significant differences between MDD patients and controls by univariate statistical analysis. A binary logistic regression (BLR) model combined four peptide markers (P1, P9, P17, P29) was established, yielding an overall prediction accuracy of 91.7% and 82.2% in the discovery and validation sets, respectively. CONCLUSIONS AND CLINICAL RELEVANCE: In conclusion, the excellent performance of the BLR model in both discovery and validation sets demonstrates the robustness of the four peptide markers panel. It is very valuable for quantification of the absolute content of four peptides and further verification.

3.
J Biomol Struct Dyn ; : 1-12, 2021 Apr 07.
Artigo em Inglês | MEDLINE | ID: mdl-33826484

RESUMO

Interaction between the SARS-COV-2 (2019 novel coronavirus) spike protein and ACE2 receptors expressed on cellular surfaces initialises viral attachment and consequent infection. Blocking this interaction shows promise for blocking or ameliorating the virus' pathological effects on the body. By contrast to work focusing on the coronavirus, which has significant potential diversity through possible accumulation of mutations during transmission, targeting the conserved ACE2 protein expressed on human cells offers an attractive alternative route to developing pharmacological prophylactics against viral invasion. In this study, we screened a virtual database of natural peptides in silico, with ACE2 as the target, and performed structural analyses of the interface region in the SARS-COV-2 RBD/ACE2 complex. These analyses have identified 15 potentially effective compounds. Analyses of ACE2/polypeptide interactions suggest that these peptides can block viral invasion of cells by stably binding in the ACE2 active site pocket. Molecular simulation results for Complestatin and Valinomycin indicate that they may share this mechanism. The discovery of this probable binding mechanism provides a frame of reference for further optimization, and design of high affinity ACE2 inhibitors that could serve as leads for production of drugs with preventive and therapeutic effects against SARS-COV-2.Communicated by Ramaswamy H. Sarma.

4.
J Sci Food Agric ; 100(14): 5152-5161, 2020 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-32529660

RESUMO

BACKGROUND: The brain is especially sensitive to diabetes-induced damage. Chronic hyperglycemia can potentially lead to brain dysfunctions, affecting spatial learning and memory. RESULTS: The type 2 diabetes (T2D) rats were administered TWK10-fermented soy milk water extract (WE) and ethanol extract (EE) for 6 weeks. WE and EE treatment attenuated T2D-induced alteration in cognitive function assessed using the Morris water maze. Moreover, administration of WE and EE significantly elevated superoxide dismutase activity (166.96% and 181.21%, P < 0.05, respectively) and reduced malondialdehyde concentration (35.03% and 43.97%, P < 0.05, respectively) in the hippocampus of the rats. Additionally, the calmodulin level and nitric oxide concentration were regulated by WE and EE. CONCLUSION: This study provides scientific evidence that WE and EE enhance anti-oxidative enzyme activity, which subsequently regulates factors associated with cognitive function in T2D rats. © 2020 Society of Chemical Industry.


Assuntos
Diabetes Mellitus Tipo 2/dietoterapia , Diabetes Mellitus Tipo 2/psicologia , Alimentos e Bebidas Fermentados/microbiologia , Lactobacillus plantarum/metabolismo , Leite de Soja/metabolismo , Animais , Cognição , Diabetes Mellitus Tipo 2/metabolismo , Diabetes Mellitus Tipo 2/fisiopatologia , Fermentação , Alimentos e Bebidas Fermentados/análise , Hipocampo/metabolismo , Hipocampo/fisiopatologia , Humanos , Masculino , Malondialdeído/metabolismo , Óxido Nítrico/metabolismo , Ratos Wistar
5.
J Agric Food Chem ; 68(7): 2263-2275, 2020 Feb 19.
Artigo em Inglês | MEDLINE | ID: mdl-31986019

RESUMO

The growth conditions and age of Panax ginseng are vital for determining the quality of the ginseng plant. However, the considerable difference in price according to the cultivation method and period of P. ginseng leads to its adulteration in the trade market. We herein focused on ginseng peptides and the possibility of these peptides to be used as biomarker(s) for discrimination of P. ginseng. We applied an ultraperformance liquid chromatography-high resolution mass spectrometry-based peptidomics approach to characterize ginseng peptides and discover novel peptide biomarkers for authentication of mountain-cultivated ginseng (MCG). We identified 52 high-confidence peptides and screened 20 characteristic peptides differentially expressed between MCG and cultivated ginseng (CG). Intriguingly, 6 differential peptides were expressed significantly in MCG and originated from dehydrins that accumulated during cold or drought conditions. In addition, 14 other differential peptides that were significantly expressed in CG derived from ginseng major protein, an essential protein for nitrogen storage. These biological associations confirmed the reliability and credibility of the differential peptides. Additionally, we determined several robust peptide biomarkers for discrimination of MCG through a precise selection process. These findings demonstrate the potential of peptide biomarkers for identification and quality control of P. ginseng in addition to ginsenoside analysis.


Assuntos
Panax/química , Peptídeos/química , Sequência de Aminoácidos , Biomarcadores/química , Cromatografia Líquida de Alta Pressão , Análise Discriminante , Contaminação de Alimentos/análise , Espectrometria de Massas , Panax/crescimento & desenvolvimento , Mapeamento de Peptídeos , Raízes de Plantas/química , Raízes de Plantas/crescimento & desenvolvimento , Controle de Qualidade
6.
AMB Express ; 9(1): 163, 2019 Oct 11.
Artigo em Inglês | MEDLINE | ID: mdl-31605256

RESUMO

Wound healing is a highly dynamic phenomenon comprising numerous coordinated steps including homeostasis/coagulation, inflammation, migration, proliferation, and remodeling. Diabetes mellitus (DM) is a multisystem chronic epidemic that prolongs inflammation in wounds and is associated with impaired healing. This study aimed to investigate the effect of an ethanol extract from Lactobacillus plantarum TWK10 (TWK10)-fermented soymilk on wound healing. The anti-inflammatory effects of the ethanol extract of TWK10-fermented soymilk on lipopolysaccharide-stimulated RAW264.7 macrophage cells were examined. The ethanol extract of TWK10-fermented soymilk (100 µg/mL) significantly decreased nitric oxide production from 11.34 ± 0.74 µM to 8.24 ± 2.02 µM (p < 0.05) and enhanced proliferation in Detroit 551 cells cultured in high-glucose medium; the cell number peaked at 128.44 ± 7.67% (compared to the untreated control) at 600 µg/mL. An ethanol extract of TWK10-fermented soymilk + vaseline-treated rat model of streptozotocin-induced diabetic wounds was generated herein, and the following groups were formed herein: normal control (NC), blank control (BC), low dose group (LD, 0.24 mg/wound), intermediate dose (MD, 0.48 mg/wound), and high dose (HD, 2.40 mg/wound). On day 14 after wound infliction, the wound area in the LD, MD, and HD groups was significantly decreased to 10.2, 8.4, and 8.5% respectively (p < 0.05). Moreover, in the LD, MD, and, HD groups, tumor necrosis factor-α, interleukin 6, and matrix metalloproteinase-9 were downregulated in the wounded skin. These results show that the topical application of the ethanol extract of TWK10-fermented soymilk is beneficial for enhancing wound healing and for the closure of diabetic wounds.

7.
J Pharm Biomed Anal ; 173: 62-67, 2019 Sep 05.
Artigo em Inglês | MEDLINE | ID: mdl-31121455

RESUMO

Measurement of peptides such as oxytocin in plasma is a critical challenge in clinical research because of their extreme low concentrations as well as the tremendous interferencing substances co-presented in plasma. In this study, we developed an efficient salt-out assisted liquid-liquid extraction (SALLE) to treat plasma, and then analyzed the samples using nano-LC-MS to quantify intact oxytocin (OT) in human and rat plasmas. Our results showed that the use of SALLE (Isopropanol/K2HPO4 (4 M)) allows efficient removal of various disrupters, including proteins, inorganic salts, and lipids, which helps avoid the risk of blocked capillary columns and matrix effects. Moreover, instant SALLE can reduce the possible binding between OT and proteins, thus allowing high repeatability of OT extraction from the original plasma. This combination of SALLE and nano-LC-MS method provided in the end a 1 pg/m L of detection limit. Comparative analysis showed that the concentration of OT in the plasma taken from 12 volunteers ranged from 3 to 214 pg/m L, about one order less than those in the plasma of rats. Compared to the previously reported LC-MS and immunoassay methods, the combination of SALLE and nano-LC-MS permits reliable measurement of intact OT even in human plasma. Our approach may be an alternative method for quantitative determination of other ultra-trace peptides in plasma, which would help the investigators understand the role of peptides in behaviours and diseases.


Assuntos
Extração Líquido-Líquido/métodos , Ocitocina/sangue , 2-Propanol/química , Animais , Cromatografia Líquida/métodos , Feminino , Voluntários Saudáveis , Humanos , Limite de Detecção , Masculino , Ocitocina/isolamento & purificação , Fosfatos/química , Compostos de Potássio/química , Ratos , Espectrometria de Massas em Tandem/métodos
8.
J Am Soc Mass Spectrom ; 30(3): 403-418, 2019 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-30644055

RESUMO

When characterizing components from ginseng, we found a vast number of multicharged anions presented in the liquid chromatography-mass spectrometry (LC-MS) chromatograms. The source of these anions is unclear yet, while ginsenosides, the major components of ginseng, are the main suspected type of molecules because of their sugar moiety. Our investigation using 14 pure ginsenosides affirmed that the multicharged anions were formed by ginsenosides rather than other types of ingredients in ginseng. Various anions could be observed for each ginsenoside. These anions contain ions ([M-2H]2-, [M+Adduct]2-), as well as those formed by polymerization of at least two ginsenosides, such as [nM-2H]2-, [nM-H+Adduct]2-, and [nM-3H]3-. The presence of so different types of ions from a ginsenoside explains the reason for the large number of anions in the LC-MS analysis of ginseng. We further found that formation of [nM-2H]2- ions was influenced by the number of sugar chains: ginsenosides containing two sugar chains produced all [nM-2H]2- ion types, whereas ginsenosides containing one sugar chain did not produce [2M-2H]2-. Thus, [2M-2H]2- and [3M-2H]2- can be utilized to rapidly identify monodesmosidic and/or bidesmosidic ginsenosides as joint diagnostic anions. The position of the glycosyl radical might be the key factor affecting the formation of multicharged multimer ions from monodesmosidic ginsenosides. Consequently, three groups of ginsenoside isomers were differentiated by characteristic [nM-2H]2- anions. Using concentration-dependent characteristics and collision-induced dissociation (CID), we confirmed that [nM-2H]2- ions are non-covalently bound multimers whose aggregation has marked distinction between monodesmosidic and bidesmosidic ginsenosides, accounting for the differentiated formation of [nM-2H]2- between them. Graphical Abstract.


Assuntos
Ginsenosídeos/análise , Ginsenosídeos/química , Espectrometria de Massas por Ionização por Electrospray/métodos , Ânions , Cromatografia Líquida , Estrutura Molecular , Panax/química , Soluções , Espectrometria de Massas em Tandem
9.
J Agric Food Chem ; 65(36): 7926-7933, 2017 Sep 13.
Artigo em Inglês | MEDLINE | ID: mdl-28829589

RESUMO

Glyceryl 1,3-dipalmitate (GD) purified from Lactobacillus paracasei subsp. paracasei NTU 101-fermented products has been demonstrated to possess neuroprotective properties. We determined the effect of GD on oxygen-glucose deprivation and reperfusion (OGD/R)-induced SH-SY5Y neuroblastoma cell death. GD ameliorated OGD/R-induced apoptosis by elevating the protein expression of nuclear peroxisome proliferator-activated receptor γ (PPARγ) and nuclear factor erythroid 2-related factor 2 (Nrf2), thereby attenuating reactive oxygen species (ROS) generation. Pretreatment with GD reduced nuclear factor κ-light-chain-enhancer of activated B cells (NF-κB) expression from 1.54 ± 0.27 to 0.84 ± 0.46, thereby attenuating the induction of pro-inflammatory mediators, and increased the plasma membrane Ca2+ ATPase (PMCA) levels from 0.81 ± 0.02 to 1.08 ± 0.06, thus reducing the levels of cytosolic Ca2+; this also correlated with reduced cell death. We conclude that GD prevents SH-SY5Y cells from injury after OGD/R insult, possibly by modulating oxidative stress and inflammatory response.


Assuntos
Glucose/metabolismo , Lactobacillus paracasei/química , Neurônios/efeitos dos fármacos , Estresse Oxidativo/efeitos dos fármacos , Oxigênio/metabolismo , Palmitatos/farmacologia , Apoptose/efeitos dos fármacos , Linhagem Celular Tumoral , Sobrevivência Celular/efeitos dos fármacos , Humanos , Lactobacillus paracasei/metabolismo , NF-kappa B/metabolismo , Neurônios/citologia , Neurônios/metabolismo , PPAR gama/genética , PPAR gama/metabolismo , Palmitatos/química , Palmitatos/metabolismo
10.
Pharm Biol ; 55(1): 487-496, 2017 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-27937042

RESUMO

CONTEXT: Numerous etiological studies have established positive clinical association between hypertension and vascular dementia (VaD). Lactobacillus paracasei subsp. paracasei NTU 101-fermented products have been shown to decrease vascular risk factors such as hypertension, atherosclerosis, hyperlipidemia and obesity. OBJECTIVE: This study investigated the effect of ethanol extract of Lactobacillus paracasei subsp. paracasei NTU 101-fermented products (NTU101F) in hypertension-induced VaD in rats. MATERIALS AND METHODS: Hypertension was promoted by subcutaneous injection of deoxycorticosterone acetate (DOCA, 25 mg/kg body weight/day, twice a week) and substitution of drinking water with 1.0% NaCl and 0.2% KCl. The NTU101F groups (0.5, 1.0, and 5.0) administered NTU101F at the concentrations 11, 22, and 110 mg/kg body weight/day, respectively, starting from day 51 day of DOCA-salt treatment. Morris water maze (MWM) was used for testing learning and memory. Different biochemical estimations were used to assess oxidative stress and inflammatory response in hippocampus. RESULTS: Oral administration of NTU101F in DOCA-salt hypertension-induced VaD rats resulted in a significant decrease in blood pressure by 18.3-23.2% (p < 0.001), which was regulated by increasing eNOS density (about 3-fold) in the aorta, promoting NO production, and decreasing of matrix metallopeptidase 9 activity (about 2-fold) in the hippocampus, in addition to improve the kidney function and structure, decrease escape latency and increase the times spent in the target quadrant by 23.5-27.8% (p < 0.05). CONCLUSION: Overall, our findings suggest that NTU101F could exert neuroprotection in the brain and attenuate hypertension-induced VaD.


Assuntos
Comportamento Animal/efeitos dos fármacos , Produtos Fermentados do Leite , Demência/prevenção & controle , Hipocampo/efeitos dos fármacos , Hipertensão/terapia , Aprendizagem em Labirinto/efeitos dos fármacos , Memória/efeitos dos fármacos , Animais , Aorta/efeitos dos fármacos , Aorta/metabolismo , Aorta/fisiopatologia , Pressão Sanguínea/efeitos dos fármacos , Citocinas/metabolismo , Demência/etiologia , Demência/fisiopatologia , Demência/psicologia , Acetato de Desoxicorticosterona , Suplementos Nutricionais , Modelos Animais de Doenças , Elastina/metabolismo , Reação de Fuga/efeitos dos fármacos , Hipocampo/metabolismo , Hipocampo/fisiopatologia , Hipertensão/induzido quimicamente , Hipertensão/metabolismo , Hipertensão/fisiopatologia , Mediadores da Inflamação/metabolismo , Rim/efeitos dos fármacos , Rim/patologia , Rim/fisiopatologia , Peroxidação de Lipídeos/efeitos dos fármacos , Masculino , Metaloproteinase 9 da Matriz/metabolismo , Óxido Nítrico/metabolismo , Óxido Nítrico Sintase Tipo III/metabolismo , Estresse Oxidativo/efeitos dos fármacos , Ratos Wistar , Fatores de Tempo
11.
Biomarkers ; 21(3): 233-42, 2016.
Artigo em Inglês | MEDLINE | ID: mdl-26846302

RESUMO

Time-course metabolic changes of aristolochic acid nephrotoxicity (AAN) was investigated using acute AAN HK-2 model. And the AAN-related biomarkers were selected. In the results, 11 potential identified biomarkers were selected and validated using multivariate method combined with time-course analysis. Several metabolic pathways, including vitamin metabolism, lipids acalytion, trytophan metabolism and protein degradation were found to be associated with AAN pathology. This research will provide a valuable reference for the discovery of more potential biomarkers of AAN progression in clinic.


Assuntos
Aristolochia/efeitos adversos , Ácidos Aristolóquicos/efeitos adversos , Biomarcadores/metabolismo , Nefropatias/metabolismo , Aristolochia/química , Ácidos Aristolóquicos/administração & dosagem , Linhagem Celular , Cromatografia Líquida de Alta Pressão , Progressão da Doença , Humanos , Nefropatias/induzido quimicamente , Nefropatias/patologia , Túbulos Renais Proximais/efeitos dos fármacos , Túbulos Renais Proximais/patologia , Metabolismo dos Lipídeos/efeitos dos fármacos , Espectrometria de Massas , Redes e Vias Metabólicas/efeitos dos fármacos , Metabolômica/métodos , Triptofano/metabolismo , Vitaminas/metabolismo
12.
Artigo em Inglês | MEDLINE | ID: mdl-26621783

RESUMO

The chemical fingerprint and metabolic profile of traditional Chinese medicine is very complicated and has been a great challenge. In the present study, chemical fingerprint of ethyl acetate fraction of Gastrodia elata (EtAcGE) and metabolic profile of rat plasma sample after intragastric administration of EtAcGE (2.5g/kg) were investigated using ultra-high performance liquid chromatography coupled with quadrupole-time of flight mass spectrometry (UPLC/Q-TOF MS). A total of 38 chemical constituents of EtAcGE were identified by comparing their retention time, accurate molecular mass and characteristic fragment ions with those of references, or tentatively characterized by comparing molecular formula, fragment ions with that of known compound or information available in literature. And 40 compounds were detected in dosed rat plasma sample, including 16 prototypes and 24 metabolites underwent metabolic process of glucuronidation, glucosylation, sulfation, methylation, hydroxylation, dehydrogenation or mixed modes. The metabolic "soft spots" was hydroxyl or carboxy group. This is the first research for chemical fingerprint and metabolic profile of EtAcGE, which lay a foundation for the further investigation of EtAcGE.


Assuntos
Cromatografia Líquida de Alta Pressão/métodos , Gastrodia/química , Espectrometria de Massas/métodos , Extratos Vegetais/sangue , Acetatos , Animais , Masculino , Extratos Vegetais/metabolismo , Extratos Vegetais/farmacocinética , Ratos , Ratos Sprague-Dawley , Reprodutibilidade dos Testes
13.
Se Pu ; 33(7): 683-90, 2015 Jul.
Artigo em Chinês | MEDLINE | ID: mdl-26672195

RESUMO

Drug-induced hepatotoxicity is a worldwide health issue. And diagnosing the injury in the early stage is still a challenge in clinic. In this study, pattern recognition analysis of the ultra high performance liquid chromatography-mass spectrometry (UPLC-MS) of hepatocytes HL7702 was performed to develop differential metabolites related to hepatotoxicity induced by hepatotoxicants, including carbon tetrachloride (CCl4), acetaminophen (APAP), emodin, aristolochic acid (AA) and triptolide. Hepatocytes injuries were induced by 48 h of treatment with CCl4 (4 mmol/L), APAP (6.5 mmol/L), emodin (14 µmol/L), AA (35 µmol/L) and triptolide (18 nmol/L), separately. Global metabolomics profiling, multivariate analysis and database searching were performed to discover common differential metabolites for live injury. The positive hepatoprotective drug, bifendate, was used to repair triptolide induced hepatocytes injury, and bifendate-induced changes of hepatotoxicity-related metabolites were investigated. In the results, fatty acid oxidation and cellular oxidative stress-related metabolites, including nicotinamide adenine dinucleotide and glutathione were significantly changed between the control and hepatotoxicant-treated groups, and after treatment with bifendate, those perturbed metabolites all partly returned to normal level. In conclusion, we discovered potential hepatotoxicity-related metabolites that could be used to evaluate hepatotoxicity induced by chemicals, drugs and traditional Chinese medicines. This study also proved that metabolomics is one of the effective tools to investigate drug-induced hepatotoxicity.


Assuntos
Doença Hepática Induzida por Substâncias e Drogas , Hepatócitos/efeitos dos fármacos , Metabolômica , Acetaminofen/toxicidade , Ácidos Aristolóquicos/toxicidade , Tetracloreto de Carbono/toxicidade , Células Cultivadas , Cromatografia Líquida de Alta Pressão , Diterpenos/toxicidade , Emodina/toxicidade , Compostos de Epóxi/toxicidade , Humanos , Espectrometria de Massas , Fenantrenos/toxicidade
14.
Se Pu ; 33(7): 699-703, 2015 Jul.
Artigo em Chinês | MEDLINE | ID: mdl-26672197

RESUMO

An ultra performance liquid chromatography-quadrupole time of flight mass spectrometry (UPLC-QTOF-MS) method was developed and validated for the determination of N6-(4-hydroxybenzyl) adenine riboside and its pharmacokinetics in rat plasma. The chromatographic conditions were optimized. The separation was performed on an Agilent ZORBAX SB-C18 column(150 mm x 3 mm, 1.8 µm) with a gradient elution of 0.2% (v/v) formic acid aqueous solution and acetonitrile as the mobile phases at a flow rate of 0.35 mL/min. The detection was accomplished in positive mode with electrospray ionization (ESI) by UPLC-QTOF-MS, and 6-benzylamino purine was used as the internal standard (IS). The results showed that the linear range of calibration curve was 0.625-160 ng/mL for N6-(4-hydroxybenzyl) adenine riboside in rat plasma with the correlation coefficient more than 0.99. The recoveries were 88.41%-108.26%. The limit of detection was 0.1 ng/mL. The intra-day and inter-day precisions (RSDs) were less than 6%, and intra-day and inter-day accuracies (REs, RE = (measured concentration-spiked concentration)/spiked concentration x 100%) were less than ±15%. The method is rapid, sensitive and accurate for the quantitation of N6-(4-hydroxybenzyl) adenine riboside, which can be used for the study of pharmacokinetics of N6-(4-hydroxybenzyl) adenine riboside.


Assuntos
Adenosina/análogos & derivados , Cromatografia Líquida de Alta Pressão , Plasma/química , Espectrometria de Massas em Tandem , Adenosina/sangue , Adenosina/farmacocinética , Animais , Ratos
15.
J Ethnopharmacol ; 176: 49-54, 2015 Dec 24.
Artigo em Inglês | MEDLINE | ID: mdl-26471288

RESUMO

ETHNOPHARMACOLOGICAL RELEVANCE: Gastrodia elata Blume, a traditional Chinese herb, was widely used against convulsant, vertigo, paralysis, epilepsy, tetanus, asthma and immune dysfunctions. Gastrodin is one of the major bioactive components of G. elata and it is known for its anticonvulsive, anti-inflammatory, antiepileptic and neuroprotective effects. MATERIALS AND METHODS: An ultra high performance liquid chromatography-fluorescence detection (UHPLC-FLD) method was developed to determine gastrodin in rat plasma. Gastrodin and Thiamphenicol (internal standard, IS) were extracted from rat plasma by immediately protein precipitation. The pharmacokinetics of gastrodin in rats by following differently administered types was studies: intragastric administration of gastrodin (100mg/kg), parishin (116 mg/kg, with the same mole of gastrodin moiety) and G. elata extract (2.3g/kg, with the same mole of gastrodin moiety). Non-compartmental pharmacokinetic profiles were constructed using the software of WinNonlin (Phoenix, version 6.3), and the pharmacokinetic parameters were compared using unpaired Student's t-test. RESULTS: The results showed that the pharmacokinetic parameters, including Cmax, Tmax, AUC0-∞, t1/2, MRT, Vd, CL, were quite different among the three types of gastrodin administration. The administration of parishin and G. elata extract, which either could convert to gastrodin in vivo or contained free gastrodin and abundant gastrodin conjugates, gave rise to higher elimination half-life (t1/2) and mean residence time (MRT) values for gastrodin compared to free gastrodin administered. CONCLUSION: The comparison of the pharmacokinetics of gastrodin among three different administered types of gastrodin in rats suggested that administration of parishin or G. elata extract in clinic may result in a longer duration time of action than that of the administration of free gastrodin. The results may provide some guidance for the clinical applications of parishin and G. elata.


Assuntos
Álcoois Benzílicos/farmacocinética , Citratos/farmacocinética , Gastrodia , Glucosídeos/farmacocinética , Extratos Vegetais/farmacocinética , Animais , Álcoois Benzílicos/administração & dosagem , Álcoois Benzílicos/sangue , Cromatografia Líquida de Alta Pressão/métodos , Citratos/administração & dosagem , Citratos/sangue , Glucosídeos/administração & dosagem , Glucosídeos/sangue , Infusões Parenterais , Extratos Vegetais/administração & dosagem , Extratos Vegetais/sangue , Ratos Sprague-Dawley , Rizoma
16.
Anal Bioanal Chem ; 407(29): 8903-10, 2015 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-26416021

RESUMO

The pharmacokinetics of parishin, gastrodin, Gastrodia elata extract and Rhizoma Gastrodiae capsule was investigated by intragastric and/or intravenous administration to rats. Parishin was metabolized into nine metabolites after intravenous administration, and the area under the curve (AUC0-∞) of parishin and its metabolites (except parishin G and parishin E) increased nonlinearly from 72.5 to 220 mg/kg. When combining regression equation with the AUC0-∞ and dose of gastrodin injection, the percent conversion of parishin to gastrodin was obtained as 50 %. Based on multi-active metabolites of parishin in vivo, integrated pharmacokinetic mode was established. It is notable that each metabolite from parishin shares the similar metabolic process at three dosages of parishin and the bioavailability of parishin was approximately 14 %. The integrated pharmacokinetic mode was successfully applied to evaluate the holistic pharmacokinetics of gastrodin injection, G. elata extract and Rhizoma Gastrodiae capsule. The results showed that the holistic pharmacokinetics of gastrodin injection and G. elata extract was closed to that of gastrodin, but for parishin and Rhizoma Gastrodiae capsule, integrated pharmacokinetic parameters were more suitable to evaluate its holistic pharmacokinetics. Graphical abstract Pharmacokinetic study of Gastrodia elata in rats.


Assuntos
Álcoois Benzílicos/sangue , Citratos/sangue , Glucosídeos/sangue , Extratos Vegetais/sangue , Administração Intravenosa , Animais , Área Sob a Curva , Álcoois Benzílicos/administração & dosagem , Álcoois Benzílicos/metabolismo , Citratos/administração & dosagem , Citratos/metabolismo , Feminino , Gastrodia/química , Glucosídeos/administração & dosagem , Glucosídeos/metabolismo , Masculino , Extratos Vegetais/administração & dosagem , Extratos Vegetais/metabolismo , Ratos , Ratos Sprague-Dawley
17.
Food Funct ; 6(11): 3522-30, 2015 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-26299532

RESUMO

The anti-obesity activity of the water extract of soy milk fermented with Lactobacillus paracasei subsp. paracasei NTU 101 (W101) was investigated. A high-fat diet (HFD) was used to induce obesity in rats, and the effects of daily W101 feeding (8 weeks) were observed. The rats fed the HFD and supplemented with low-dose W101 (LW101, 15 mg per kg body weight per day) or high-dose W101 (HW101, 150 mg per kg body weight per day) had significantly reduced final body weight in comparison with that of the HFD group. W101 decreased the formation of lipid plaques in the aorta, reduced the adipocyte cross-sectional area and diameter, and reduced the levels of CCAAT/enhancer-binding protein ß (C/EBPß), peroxisome proliferator associated receptor γ (PPARγ), and C/EBPα. Regarding lipogenesis regulation in adipocytes, W101 suppressed heparin-releasable lipoprotein lipase (HR-LPL) in adipose tissues and inhibited lipid absorption, thereby reducing lipogenesis. Lactobacillus paracasei subsp. paracasei NTU 101-fermented soy milk may be used to develop health foods that prevent obesity.


Assuntos
Fármacos Antiobesidade/farmacologia , Produtos Fermentados do Leite/química , Obesidade/tratamento farmacológico , Leite de Soja/química , Água , Adipócitos/efeitos dos fármacos , Animais , Peso Corporal/efeitos dos fármacos , Proteína alfa Estimuladora de Ligação a CCAAT/metabolismo , Proteína beta Intensificadora de Ligação a CCAAT/metabolismo , Produtos Fermentados do Leite/microbiologia , Dieta Hiperlipídica/efeitos adversos , Gorduras na Dieta/efeitos adversos , Lactobacillus/metabolismo , Lipogênese/efeitos dos fármacos , Lipase Lipoproteica/metabolismo , Masculino , Obesidade/etiologia , PPAR gama/metabolismo , Ratos , Ratos Sprague-Dawley
18.
J Biochem Mol Toxicol ; 29(11): 533-43, 2015 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-26149242

RESUMO

The present study was undertaken to investigate the metabolic responses of human liver cells HL-7702 on chemicals purified from traditional Chinese medicine: emodin, triptolide, and aristolochic acid. Cytotoxicity tests demonstrated a dose-dependent toxic effect of emodin, triptolide, and aristolochic acid on HL7702 cells for 48 h. Emodin (14 µM), aristolochic acid (12 µg/mL), or triptolide (18 nM) was individually administrated to HL7702 and cell samples were collected after 48 h for metabolites extraction and analysis. Pattern recognition analysis reflected the significant difference in metabolic profiles between chemical-treated groups and the control group. Finally, eight metabolites including N1 -acetylspermidine, Glu Gly, N-undecanoylglycine, C16 sphinganine, sphinganine, glutathione, l-palmitoylcarnitine, and elaidic carnitine were detected as potential common biomarkers. Three pathways including sphinganine metabolism, fatty acid oxidation, and oxidative stress were identified. Our findings indicated that metabolomics would be an efficient approach to understand the molecular mechanism of hepatotoxicity induced by chemicals.


Assuntos
Ácidos Aristolóquicos/farmacologia , Diterpenos/farmacologia , Emodina/farmacologia , Hepatócitos/efeitos dos fármacos , Medicina Tradicional Chinesa , Metabolômica , Fenantrenos/farmacologia , Linhagem Celular , Cromatografia Líquida , Compostos de Epóxi/farmacologia , Hepatócitos/metabolismo , Humanos , Espectrometria de Massas
19.
Biomed Chromatogr ; 29(12): 1913-20, 2015 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-26010793

RESUMO

Parishin is a dominant active ingredient originating from Gastrodia elata Blume, and has good neuroprotective effects against brain disorders. In the present study, the metabolic profile of parishin by in vitro and in vivo experiments was investigated using ultra-high performance liquid chromatography coupled with quadrupole-time of flight mass spectrometry (UHPLC/Q-TOF MS) combined with an automated MS(E) technique. By comparison with reference compounds, accurate mass measurement, the characteristic fragmentation patterns of the parent drug parishin and gastrodin and relevant bio-transformation knowledge, 14 metabolites (seven hydrolyzates and seven derivatives of gastrodin) were detected and identified in rat plasma and urine after intragastric administration of parishin, including processes of hydrolyzation, oxidation, sulfation and glucuronidation. According to the proposed metabolic pathways of parishin, in vitro hydrolytic experiments and metabolic study of gastrodin in rat plasma, it can be inferred that parishin mainly functions as a prodrug and undergoes hydrolysis before being absorbed into the blood. The hydrolyzate, mainly gastrodin, was involved in further metabolism, which was responsible for pharmacological activities of parishin. In conclusion, this work provides valuable information on parishin metabolism using a rapid and reliable UHPLC/Q-TOF MS method, which could be widely used for the metabolic investigation of natural product.


Assuntos
Álcoois Benzílicos , Cromatografia Líquida de Alta Pressão/métodos , Citratos , Glucosídeos , Espectrometria de Massas/métodos , Animais , Álcoois Benzílicos/sangue , Álcoois Benzílicos/química , Álcoois Benzílicos/metabolismo , Álcoois Benzílicos/urina , Citratos/administração & dosagem , Citratos/sangue , Citratos/química , Citratos/metabolismo , Citratos/urina , Glucosídeos/administração & dosagem , Glucosídeos/sangue , Glucosídeos/química , Glucosídeos/metabolismo , Glucosídeos/urina , Glucuronídeos , Masculino , Ratos , Ratos Sprague-Dawley , Sulfatos
20.
Mol Biosyst ; 11(2): 635-46, 2015 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-25474166

RESUMO

Estradiol is a major drug used clinically to alleviate osteoporosis, partly through inhibition of the activity of osteoclasts, which play a crucial role in bone resorption. So far, little is known about the effects of estradiol on osteoclast metabolism. In this study, ultra-high performance liquid chromatography-tandem mass spectrometry (UPLC/MS)-based metabolomics strategy was used to investigate the metabolite response to 17ß-estradiol in mouse osteoclast RAW264.7, a commonly used cell model for studying osteoporosis. Our results showed that the application of estradiol altered the levels of 27 intracellular metabolites, including lysophosphatidylcholines (LysoPCs), other lipids and amino acid derivants. The changes of all the 27 metabolites were observed in the study of estradiol induced osteoclast proliferation inhibition (1 µM estradiol applied), while the changes of only 18 metabolites were observed in the study of differentiation inhibition (0.1 µM estradiol applied). Further pathway impact analysis determined glycerophospholipid metabolism as the main potential target pathway of estradiol, which was further confirmed by LCAT (phosphatidylcholine-sterol acyltransferase) activity changes and lipid peroxidative product (MDA, methane dicarboxylic aldehyde) changes caused by estradiol. Additionally, we found that estradiol significantly decreased intracellular oxidative stress during cell proliferation but not during cell differentiation. Our study suggested that estradiol generated a highly condition-dependent influence on osteoclast metabolism.


Assuntos
Diferenciação Celular/efeitos dos fármacos , Estradiol/farmacologia , Metabolômica/métodos , Osteoclastos/citologia , Osteoclastos/metabolismo , Animais , Proliferação de Células/efeitos dos fármacos , Cromatografia Líquida de Alta Pressão , Cromatografia de Fase Reversa , Confiabilidade dos Dados , Glicerofosfolipídeos/metabolismo , Espaço Intracelular/metabolismo , Malondialdeído/metabolismo , Metaboloma/efeitos dos fármacos , Camundongos , Análise Multivariada , Osteoclastos/efeitos dos fármacos , Osteoclastos/enzimologia , Estresse Oxidativo/efeitos dos fármacos , Fosfatidilcolina-Esterol O-Aciltransferase/metabolismo , Células RAW 264.7 , Superóxido Dismutase/metabolismo
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