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1.
BMC Complement Med Ther ; 21(1): 41, 2021 Jan 21.
Artigo em Inglês | MEDLINE | ID: mdl-33478471

RESUMO

BACKGROUND: The latest coronavirus SARS-CoV-2, discovered in China and rapidly spread Worldwide. COVID-19 affected millions of people and killed hundreds of thousands worldwide. There are many ongoing studies investigating drug(s) suitable for preventing and/or treating this pandemic; however, there are no specific drugs or vaccines available to treat or prevent SARS-CoV-2 as of today. METHODS: Fifty-eight fragrance materials, which are classified as allergen fragrance molecules, were selected and used in this study. Docking simulations were carried out using four functional proteins; the Covid19 Main Protase (MPro), Receptor binding domain (RBD) of spike protein, Nucleocapsid, and host Bromodomain protein (BRD2), as target macromolecules. Three different software, AutoDock, AutoDock Vina (Vina), and Molegro Virtual Docker (MVD), running a total of four different docking protocol with optimized energy functions were used. Results were compared with the five molecules reported in the literature as potential drugs against COVID-19. Virtual screening was carried out using Vina, molecules satisfying our cut-off (- 6.5 kcal/mol) binding affinity was confirmed by MVD. Selected molecules were analyzed using the flexible docking protocol of Vina and AutoDock default settings. RESULTS: Ten out of 58 allergen fragrance molecules were selected for further docking studies. MPro and BRD2 are potential targets for the tested allergen fragrance molecules, while RBD and Nucleocapsid showed weak binding energies. According to AutoDock results, three molecules, Benzyl Cinnamate, Dihydroambrettolide, and Galaxolide, had good binding affinities to BRD2. While Dihydroambrettolide and Galaxolide showed the potential to bind to MPro, Sclareol and Vertofix had the best calculated binding affinities to this target. When the flexible docking results analyzed, all the molecules tested had better calculated binding affinities as expected. Benzyl Benzoate and Benzyl Salicylate showed good binding affinities to BRD2. In the case of MPro, Sclareol had the lowest binding affinity among all the tested allergen fragrance molecules. CONCLUSION: Allergen fragrance molecules are readily available, cost-efficient, and shown to be safe for human use. Results showed that several of these molecules had comparable binding affinities as the potential drug molecules reported in the literature to target proteins. Thus, these allergen molecules at correct doses could have significant health benefits.


Assuntos
Alérgenos/química , Alérgenos/imunologia , /imunologia , Odorantes , Perfumes/química , /imunologia , Monofosfato de Adenosina/análogos & derivados , Monofosfato de Adenosina/química , Monofosfato de Adenosina/metabolismo , Alanina/análogos & derivados , Alanina/química , Alanina/metabolismo , Alérgenos/administração & dosagem , Alérgenos/uso terapêutico , Benzopiranos/química , Benzopiranos/metabolismo , Compostos de Benzil/química , Compostos de Benzil/metabolismo , Cinamatos/química , Cinamatos/metabolismo , /metabolismo , /metabolismo , Diterpenos/química , Diterpenos/metabolismo , Avaliação Pré-Clínica de Medicamentos , Humanos , Ligantes , Simulação de Acoplamento Molecular , Perfumes/administração & dosagem , Perfumes/uso terapêutico , Fosfoproteínas/química , Fosfoproteínas/metabolismo , Glicoproteína da Espícula de Coronavírus/química , Glicoproteína da Espícula de Coronavírus/metabolismo , Fatores de Transcrição/química , Fatores de Transcrição/metabolismo
2.
Gene ; 773: 145417, 2021 Mar 20.
Artigo em Inglês | MEDLINE | ID: mdl-33444679

RESUMO

Melissa officinalis (lemon balm) is a well-known pharmaceutical plant in traditional medicine around the world because of the high-value secondary metabolites. Nowadays, advances in computational biology and bioinformatics have opened new avenues to plant-based natural product drug discovery. Despite the pharmacological importance, there is low information about the genes encoding the important biosynthetic pathways related to the secondary metabolite in M. officinalis. In this study, the main genes related to the rosmarinic acid (RA) and terpenoid biosynthesis pathways were detected using transcriptome analysis. Furthermore, we isolated and characterized a novel M. officinalis Hydroxyphenylpyruvate reductase (HPPR) gene involved in RA biosynthesis pathway. An effective pipeline was used to generate 37,055 unigenes by evaluating 42,837,601 Illumina paired-end reads. Functional annotation of the unigenes revealed that 27,363 (73.84%) and 35,822 (96.67%) unigenes had significant similarity to identified proteins in the SwissProt and NR databases, respectively. Also, 10,062 (36.83%) out of 37,055 unigenes were assigned to 399 KEGG pathways. Since terpenes and RA are two prominent metabolites in this plant, the attention of this study has been on the pathways related to them. A total of 149 unigenes were found that are related to the terpenoids biosynthesis, including 75 unigenes involved in the methyl-erythritol phosphate and mevalonate pathway, terpenoid backbone biosynthesis genes, and 74 unigenes related to the terpene synthase. We also identified 144 and 30 unigenes that were associated with the biosynthesis of phenylpropanoid and the rosmarinic acid pathway. Consequently, this investigation can be a comprehensive and accurate transcriptome basis for further investigation in the metabolic engineering and detection of new genes and pathways in M. officinalis.


Assuntos
Cinamatos/metabolismo , Depsídeos/metabolismo , Melissa/genética , Terpenos/metabolismo , Transcriptoma/genética , Alquil e Aril Transferases/genética , Alquil e Aril Transferases/metabolismo , Vias Biossintéticas/genética , Biologia Computacional , Perfilação da Expressão Gênica , Regulação da Expressão Gênica de Plantas/genética , Sequenciamento de Nucleotídeos em Larga Escala , Melissa/metabolismo , Anotação de Sequência Molecular , Folhas de Planta/genética , Folhas de Planta/metabolismo
3.
PLoS One ; 15(9): e0239019, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-32941497

RESUMO

The melanosome is a specialized membrane-bound organelle that is involved in melanin synthesis, storage, and transportation. In contrast to melanosome biogenesis, the processes underlying melanosome degradation remain largely unknown. Autophagy is a process that promotes degradation of intracellular components' cooperative process between autophagosomes and lysosomes, and its role for process of melanosome degradation remains unclear. Here, we assessed the regulation of autophagy and its contributions to depigmentation associated with Melasolv (3,4,5-trimethoxycinnamate thymol ester). B16F1 cells-treated with Melasolv suppressed the α-MSH-stimulated increase of melanin content and resulted in the activation of autophagy. However, introduction of bafilomycin A1 strongly suppressed melanosome degradation in Melasolv-treated cells. Furthermore, inhibition of autophagy by ATG5 resulted in significant suppression of Melasolv-mediated depigmentation in α-MSH-treated cells. Taken together, our results suggest that treatment with Melasolv inhibits skin pigmentation by promoting melanosome degradation via autophagy activation.


Assuntos
Cinamatos/farmacologia , Melanossomas/efeitos dos fármacos , Melanossomas/metabolismo , Animais , Autofagossomos/metabolismo , Autofagia/efeitos dos fármacos , Linhagem Celular Tumoral , Cinamatos/metabolismo , Macrolídeos/farmacologia , Melaninas/metabolismo , Melanócitos/metabolismo , Camundongos , Pigmentação/efeitos dos fármacos , Transtornos da Pigmentação/metabolismo , Pigmentação da Pele/efeitos dos fármacos , alfa-MSH/efeitos dos fármacos , alfa-MSH/metabolismo
4.
J Chromatogr A ; 1620: 461003, 2020 Jun 07.
Artigo em Inglês | MEDLINE | ID: mdl-32156458

RESUMO

The enormous growth in drug discovery paradigm has necessitated continuous exploration of new methods for drug-protein interaction analysis. To enhance the role of these methodologies in designing rational drugs, this work extended an immobilized angiotensin II type I receptor (AT1R) based affinity chromatography in antihypertensive compound identification. We fused haloalkane dehalogenase at C-terminus of AT1R and expressed the fusion receptor in E. coli. The expressed receptor was covalently immobilized onto 8.0 µm microspheres by mixing the cell lysate with 6-chlorocaproic acid-modified amino polystyrene microspheres. The immobilized AT1R was utilized for thermodynamic and kinetic interaction analysis between the receptor and four specific ligands. Following confirmation of these interactions by molecular docking, we identified puerarin and rosmarinic acid by determining their binding to the receptor. Azilsartan, candesartan, valsartan and olmesartan displayed two kinds of binding sites to AT1R by injection amount-dependent method. By molecular docking, we recognize the driving forces of the interaction as electrostatic interaction, hydrogen bonds and van der Waals force. The dissociation rate constants (kd) of azilsartan, candesartan, valsartan and olmesartan to AT1R were 0.01138 ± 0.003, 0.05142 ± 0.003, 0.07547 ± 0.004 and 0.01310 ± 0.005 min-1 by peak profiling assay. Comparing with these parameters, puerarin and rosmarinic acid presented lower affinity (KA: 0.12 × 104 and 1.5 × 104/M) and slower kinetics (kd: 0.6864 ± 0.03 and 0.3005 ± 0.01 min-1) to the receptor. These results, taking together, indicated that the immobilized AT1R has the capacity to probe antihypertensive compounds.


Assuntos
Anti-Hipertensivos/metabolismo , Ensaios de Triagem em Larga Escala/métodos , Receptor Tipo 1 de Angiotensina/metabolismo , Anti-Hipertensivos/química , Benzimidazóis/química , Benzimidazóis/metabolismo , Sítios de Ligação , Cromatografia de Afinidade , Cinamatos/metabolismo , Depsídeos/metabolismo , Imidazóis/química , Imidazóis/metabolismo , Proteínas Imobilizadas/química , Proteínas Imobilizadas/metabolismo , Isoflavonas/metabolismo , Cinética , Ligantes , Simulação de Acoplamento Molecular , Oxidiazóis/química , Oxidiazóis/metabolismo , Receptor Tipo 1 de Angiotensina/química , Receptor Tipo 1 de Angiotensina/genética , Proteínas Recombinantes de Fusão/química , Proteínas Recombinantes de Fusão/metabolismo , Tetrazóis/química , Tetrazóis/metabolismo , Termodinâmica , Valsartana/química , Valsartana/metabolismo
5.
Molecules ; 25(4)2020 Feb 14.
Artigo em Inglês | MEDLINE | ID: mdl-32075047

RESUMO

Advances in cancer treatment have led to significant improvements in long-term survival in many types of cancer, but heart dysfunction and heart failure, associated with cancer treatment, have also increased. Anthracyclines are the main cause of this type of cardiotoxicity. In this study, we describe a combined experimental and cell morphology analysis approach for the high-throughput measurement and analysis of a cardiomyocyte cell profile, using partial least square linear discriminant analysis (PLS-LDA) as the pattern recognition algorithm. When screening a small-scale natural compound library, rosmarinic acid (RosA), as a candidate drug, showed the same cardioprotective effect as the positive control. We investigated the protective mechanism of RosA on a human cardiomyocyte cell line (AC16) and human induced pluripotent stem-cell-derived cardiomyocytes (hiPSC-CMs). We showed that RosA pretreatment suppressed doxorubicin (Dox)-induced cell apoptosis and decreased the activity of caspase-9. RosA promotes the expression of Heme oxygenase-1 (HO-1) and reduces the production of reactive oxygen species (Ros), which is induced by Dox. Meanwhile, it can also promote the expression of cardiac-development-related protein, including histone deacetylase 1 (HDAC1), GATA binding protein 4 (GATA4) and troponin I3, cardiac type (CTnI). Collectively, our data support the notion that RosA is a protective agent in hiPSC-CMs and has the potential for therapeutic use in the treatment of cancer therapy-related cardiac dysfunction and heart failure.


Assuntos
Cardiotoxicidade/metabolismo , Cinamatos/metabolismo , Depsídeos/metabolismo , Doxorrubicina/efeitos adversos , Miócitos Cardíacos/efeitos dos fármacos , Antibióticos Antineoplásicos/efeitos adversos , Antibióticos Antineoplásicos/farmacologia , Apoptose/efeitos dos fármacos , Cardiotoxicidade/etiologia , Cardiotoxicidade/patologia , Diferenciação Celular/efeitos dos fármacos , Proliferação de Células/efeitos dos fármacos , Células Cultivadas , Doxorrubicina/farmacologia , Humanos , Células-Tronco Pluripotentes Induzidas/efeitos dos fármacos , Miócitos Cardíacos/metabolismo , Neoplasias/complicações , Neoplasias/tratamento farmacológico
6.
Food Chem ; 317: 126415, 2020 Jul 01.
Artigo em Inglês | MEDLINE | ID: mdl-32087518

RESUMO

This paper focused on improving antityrosinase ability of quercetin, cinnamic acid, and ferulic acid (named Q-CA-FA) from Asparagus by combining heating with ultrasound treatments. Fluorescence spectroscopy and UPLC-MS were used to evaluate inhibitory mechanisms. Results showed that the impacts of combining heating (150 °C for 30 min) with ultrasound (600 W for 30 min) treatments were similar to heating treatment (150 °C for 120 min) alone, and the inhibition rate could reach 98.2% in the addition of 5 mM Q-CA-FA. Fluorescence quenching indicated that treated Q-CA-FA-tyrosinase complex was more stable, but combining treatments did not change the major force between tyrosinase and polyphenols. Thermodynamic analysis illustrated that the randomness of compounds was also increased. Interestingly, 2-hydroxy-3-(3-hydroxy-4-methoxy-phenyl)-propionic acid 4-(2,3-dihydroxy-propyl)-phenyl ester was newly detected, which might be the major reason for enhancing antityrosinase ability. Taken together, these results provide a creative insight on increasing antityrosinase activity by combining heating with ultrasound treatments.


Assuntos
Monofenol Mono-Oxigenase/metabolismo , Polifenóis/metabolismo , Sonicação , Asparagus (Planta)/química , Asparagus (Planta)/metabolismo , Cromatografia Líquida de Alta Pressão , Cinamatos/análise , Cinamatos/metabolismo , Ácidos Cumáricos/análise , Ácidos Cumáricos/metabolismo , Temperatura Alta , Monofenol Mono-Oxigenase/antagonistas & inibidores , Polifenóis/análise , Quercetina/análise , Quercetina/metabolismo , Espectrometria de Fluorescência , Espectrometria de Massas em Tandem , Termodinâmica
7.
Biomed Chromatogr ; 34(4): e4806, 2020 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-32012312

RESUMO

Rosmarinic acid (RA) is a phenolic acid originally isolated from the herb medicine Rosmarinus officinalis. The purpose of this study was to identify the metabolites of RA. RA was incubated with human liver microsomes in the presence of ß-nicotinamide adenine dinucleotide phosphate tetrasodium salt and/or uridine diphosphate glucuronic acid using glutathione (GSH) as a trapping agent. After 60-min incubation, the samples were analyzed using high-resolution liquid chromatography tandem mass spectrometry. Under the current conditions, 14 metabolites were detected and identified. Our data revealed that RA was metabolized through the following pathways: the first pathway is the oxidation of catechol to form ortho-quinone intermediates, which react with GSH to form mono-GSH adducts (M1, M2, and M3) and bis-GSH adducts (M4 and M5); the second pathway is conjugation with glucuronide to yield acylglucuronide (M7), which further reacts with GSH to form RA-S-acyl-GSH adduct (M9); the third pathway is hydroxylation to form M10, M11, and M12, which further react with GSH to form mono-GSH adducts (M13 and M14); the fourth pathway is conjugation with GSH through Michael addition (M6); the fifth pathway is conjugation with glucuronidation, forming M8, which is the major metabolic pathway of RA.


Assuntos
Cromatografia Líquida/métodos , Cinamatos/metabolismo , Depsídeos/metabolismo , Microssomos Hepáticos/metabolismo , Espectrometria de Massas em Tandem/métodos , Glutationa , Humanos , Modelos Moleculares , NADP , Espectrometria de Massas por Ionização por Electrospray , Uridina Difosfato Ácido Glucurônico
8.
Biomolecules ; 10(2)2020 01 24.
Artigo em Inglês | MEDLINE | ID: mdl-31991557

RESUMO

The present study demonstrates hormonal control of Salvia viridis growth and development using four different purine-type cytokinins at different concentrations. The addition of cytokinins significantly increased biomass of cultures, proliferation rate, and, interestingly, secondary metabolite production. The best response in terms of multiplication ratio was recorded on Murashige and Skoog medium supplemented with 0.5 mg/L BPA (N-benzylotetrahydropyranyl adenine), while the greatest biomass accumulation was achieved when supplemented with 1 mg/L m-T (meta-topoline). Quantitative UPLC-DAD analysis of the hydromethanolic extract from S. viridis culture revealed the presence of 12 polyphenols: seven phenolic acids and five phenylethanoids. The highest total content of polyphenolic compounds was found in shoots cultivated on medium with 2 mg/L BPA (18.66 mg/g DW): almost twice that of control shoots. The medium was also the most optimal for the biosynthesis of rosmarinic acid, the predominant phenolic acid. However, the greater phenylethanoid accumulation was stimulated by 1 mg/L m-T: the metabolite content was above three times higher than that found in shoots grown on the control medium (8.03 mg/g DW vs. 2.37 mg/g DW). Hence, it was demonstrated that phytohormones are capable of influencing not only vital physiological processes, but therapeutic potential of plants as well. Therefore, the cytokinin-based sage cultures may be also considered as the alternative sources of bioactive compounds.


Assuntos
Proliferação de Células/efeitos dos fármacos , Citocininas/farmacologia , Purinas/farmacologia , Salvia/efeitos dos fármacos , Cinamatos/metabolismo , Depsídeos/metabolismo , Hidroxibenzoatos/metabolismo , Células Vegetais/efeitos dos fármacos , Brotos de Planta/efeitos dos fármacos , Brotos de Planta/crescimento & desenvolvimento , Polifenóis/biossíntese , Salvia/crescimento & desenvolvimento
9.
Biochimie ; 170: 128-139, 2020 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-31945397

RESUMO

In search of the compounds that interfere with amyloid transformation of alpha-synuclein, 9 natural and synthetic cinnamic acid derivatives were studied. They are structurally similar to a half of curcumin, which has pronounced anti-aggregatory and anti-amyloid effects. We have shown that some of these derivatives prevent ovine prion protein amyloidization. Subsequently, thioflavin T binding assay showed that 3 out of 9 studied compounds effectively prevented amyloid transformation of alpha-synuclein with IC50 of 13, 50 and 251 µM. Molecular modeling approach revealed possible binding sites of the three selected ligands with alpha-synuclein fibrils, while monomeric alpha-synuclein does not bind to the ligands according to experimental results. This led us to believe that compounds may act by changing the structure of primary aggregates, preventing the formation of full-length fibrils. The inhibiting effect of the ligands on aggregation of alpha-synuclein was further confirmed by monitoring aggregation via turbidimetry, susceptibility to proteolytic cleavage, changes in beta-sheet content, and scanning ion-conductance microscopy. Studied derivatives were not cytotoxic, and, moreover, two studied compounds (ferulic and 3,4-dimethoxycinnamic acid) are found in plant sources and are natural metabolites present in human blood, so they can be promising candidate drugs for synucleinopathies, including Parkinson's disease.


Assuntos
Amiloide/química , Amiloide/metabolismo , Produtos Biológicos/metabolismo , Cinamatos/metabolismo , alfa-Sinucleína/química , alfa-Sinucleína/metabolismo , Produtos Biológicos/química , Cinamatos/química , Humanos , Simulação de Acoplamento Molecular , Conformação Proteica
10.
J Sci Food Agric ; 100(6): 2410-2417, 2020 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-31917463

RESUMO

BACKGROUND: Greek oregano is a culinary and medicinal herb native to the Mediterranean region; however nowadays it is cultivated in many regions all over the world. It is commonly used as a spice for flavoring food products and in various traditional medicine applications. This study investigated the effect of nitrogen fertilization at 0-150 kg N ha-1 on the potential bioaccessibility and activity of Greek oregano compounds. For this purpose the total phenolic content, rosmarinic acid content, and antioxidant activities of the raw material, as well as digested fractions, were determined. RESULTS: Nitrogen fertilization had a negative influence on the phenolic content and antioxidant activity of raw material; however, its effect on the potential bioaccessibility varied depending on the dose. The highest potential bioaccessibility and activity was determined for plants fertilized with 30 kg N ha-1 . For the gastric and intestinal phases of digestion, the potential bioaccessibility percentages were 39.5% and 29.6% for total phenolics, 53.1% and 11.2% for rosmarinic acid content, 45.2% and 44.4% for antiradical activity against 2,2'-azino-bis (3-ethylbenzothiazoline-6-sulfonic acid (ABTS), 39.2% and 27.2% for antiradical activity against 2,2-diphenyl-1-picrylhydrazyl (DPPH) and 50.2% and 23.4% for reducing power, respectively. CONCLUSION: The results showed that nitrogen fertilization is an important factor determining the in vitro bioaccessibility of Greek oregano compounds. Furthermore, the nutraceutical potential of herbs, in term of the bioaccessibility of bioactive compounds, may be optimized during plant cultivation by applying an accurate nitrogen level. © 2020 Society of Chemical Industry.


Assuntos
Fertilizantes , Nitrogênio , Origanum/química , Antioxidantes/análise , Antioxidantes/metabolismo , Disponibilidade Biológica , Cinamatos/análise , Cinamatos/metabolismo , Depsídeos/análise , Depsídeos/metabolismo , Digestão , Origanum/metabolismo , Fenóis/análise , Fenóis/metabolismo
11.
Biochem J ; 477(1): 61-74, 2020 01 17.
Artigo em Inglês | MEDLINE | ID: mdl-31764941

RESUMO

Black pepper, dried green fruit of Piper nigrum L., is a household spice most popular in the world. Piperine, the pungency compound of black pepper, is proposed to partially arise from phenylpropanoid pathway. In the biosynthesis of piperine, 4-coumarate:CoA ligase (4CLs) must play a pivotal role in activating intermediate acids to corresponding CoA thioesters to serve as substrates. Based on transcriptome data, we isolated three P. nigrum 4CL isoforms (Pn4CL1, -2, and -3) from unripe peppercorn. These Pn4CLs were expressed in E. coli for in vitro enzyme assay with putative substrates, namely cinnamic, coumaric, ferulic, piperonylic, 3,4-methylenedioxycinnamic (3,4-MDCA), and piperic acids. Phylogenetic analysis and substrate usage study indicated that Pn4CL1, active towards coumaric and ferulic acids, belongs to class I 4CL for lignin synthesis. Pn4CL2 was a typical cinnamate-specific coumarate:CoA ligase-like (CLL) protein. The Pn4CL3, as class II enzyme, exhibited general 4CL activity towards coumaric and ferulic acids. However, Pn4CL3 was also active towards piperonylic acid, 3,4-MDCA, and piperic acid. Pn4CL3 possessed ∼2.6 times higher catalytic efficiency (kcat/KM) towards 3,4-MDCA and piperic acid than towards coumaric and ferulic acids, suggesting its specific role in piperine biosynthesis. Different substrate preference among the Pn4CL isoforms can be explained by 3-dimensional protein structure modeling, which demonstrated natural variants in amino acid residues of binding pocket to accommodate different substrates. Quantitative PCR analysis of these isoforms indicated that Pn4CL1 transcript level was highest in the roots whereas Pn4CL2 in the fruits and Pn4CL3 in the leaves.


Assuntos
Cinamatos/metabolismo , Coenzima A Ligases/química , Ácidos Graxos Insaturados/biossíntese , Piper nigrum/enzimologia , Frutas/enzimologia , Isoenzimas/química , Folhas de Planta/enzimologia , Raízes de Plantas/enzimologia , Especificidade por Substrato
12.
Environ Toxicol Chem ; 39(3): 574-586, 2020 03.
Artigo em Inglês | MEDLINE | ID: mdl-31749247

RESUMO

The present study investigated the dietary bioaccumulation and biotransformation of hydrophobic organic sunscreen agents, 2-ethylhexyl-4-methoxycinnamate (EHMC) and octocrylene (OCT), in rainbow trout using a modified Organisation for Economic Co-operation and Development 305 dietary bioaccumulation test that incorporated nonbiotransformed reference chemicals. Trout were exposed to 3 dietary concentrations of each chemical to investigate the relationship between dietary exposure concentration and observed accumulation and depuration. Both EHMC and OCT were significantly biotransformed, resulting in mean in vivo whole-body biotransformation rate constants (kMET ) of 0.54 ± 0.06 and 0.09 ± 0.01 d-1 , respectively. The kMET values generated for both chemicals did not differ between dietary exposure concentrations, indicating that chemical concentrations in the fish were not high enough to saturate biotransformation enzymes. Both somatic and luminal biotransformation substantially reduce EHMC and OCT bioaccumulation potential in trout. Biomagnification factors (BMFs) and bioconcentration factors (BCFs) of EHMC averaged 0.0035 kg lipid kg lipid-1 and 396 L kg-1 , respectively, whereas those of OCT averaged 0.0084 kg lipid kg lipid-1 and 1267 L kg-1 . These values are 1 to 2 orders of magnitude lower than the BMFs and BCFs generated for reference chemicals of similar log KOW . In addition, for both chemicals, derived BMFs and BCFs fell below established bioaccumulation criteria (1.0 kg lipid kg lipid-1 and 2000 L kg-1 , respectively), suggesting that EHMC ad OCT are unlikely to bioaccumulate to a high degree in aquatic biota. Environ Toxicol Chem 2020;39:574-586. © 2019 SETAC.


Assuntos
Acrilatos/metabolismo , Bioacumulação , Cinamatos/metabolismo , Oncorhynchus mykiss/metabolismo , Protetores Solares/metabolismo , Poluentes Químicos da Água/metabolismo , Animais , Biotransformação , Interações Hidrofóbicas e Hidrofílicas
13.
Biomed Chromatogr ; 34(3): e4773, 2020 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-31813160

RESUMO

Lonicerae Japonicae Flos (LJF) is a typical herbal medicine and is used as a functional food. LJF, which has complex chemical compounds, has various biological effects. The global metabolomics, focusing on both the endogenous and exogenous metabolites, have not yet been investigated for LJF in normal healthy rats using LC-MS. In this study, plasma metabolomics was analyzed after the administration of LJF at different time intervals, and the exogenous metabolites were identified. Partial least squares discriminant analysis showed significant differences in chemical content in the dosed rats. Cholic acid, indoleacrylic acid, indolelactic acid, hippuric acid, N-acetyl-phenylalanine, and N-acetyl-serotonin significantly accumulated in the dosed rats. Lysophosphatidylethanolamine and lysophosphatidylcholine content, including plasmalogen, increased. There were 25 components of LJF, including 15 prototypes and 10 metabolites, that were identified. The 15 prototypes included phenolic acids, flavonoids, and iridoids, and their contents decreased with an increase in the administration time. Glucuronidation and sulfation of polyphenols were found for LJF. The exogenous glucuronide and sulfate metabolites-including dihydrocoumaric acid-sulfate, dihydrocaffeic acid-sulfate, dihydroferulic acid-sulfate, apigenin-glucuronide, apigenin-glucuronide-sulfate, isorhamnetin-glucuronide-sulfate, and others-were identified with a neutral loss of 176 and 80, respectively. The metabolic differences found in the study may serve as biomarkers of LJF consumption and promote the understanding of the mechanism of action of LJF.


Assuntos
Lonicera , Metaboloma/efeitos dos fármacos , Metabolômica/métodos , Extratos Vegetais , Administração Oral , Animais , Biomarcadores/sangue , Biomarcadores/química , Biomarcadores/metabolismo , Cromatografia Líquida de Alta Pressão , Cinamatos/sangue , Cinamatos/química , Cinamatos/metabolismo , Masculino , Espectrometria de Massas , Extratos Vegetais/administração & dosagem , Extratos Vegetais/metabolismo , Extratos Vegetais/farmacologia , Quercetina/sangue , Quercetina/química , Quercetina/metabolismo , Ratos , Ratos Sprague-Dawley , Reprodutibilidade dos Testes
14.
Plant Cell Rep ; 39(2): 207-215, 2020 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-31713663

RESUMO

KEY MESSAGE: Benzoate-Coenzyme A ligase enzyme activity catalyzing the conversion of free benzoic acid to benzoyl-CoA was detected and biochemically characterized in the elicitor-treated pear cell cultures. Asian pear (Pyrus pyrifolia) is an economically and nutritionally important fruit-bearing tree of the subtribe Malinae. Upon pathogen attack, pears produce unique benzoate-derived biphenyl phytoalexins. The upstream biosynthesis of the biphenyl in Malinae is still incomplete. Previously, protein preparations from yeast extract-treated pear cultures were able to convert L-phenylalanine to cinnamic acid catalyzed by the activity of the phenylalanine ammonia lyase. The same extract was able to perform a C2 side-chain cleavage of cinnamic acid to benzaldehyde followed by oxidation of the latter to benzoic acid owing to the molecularly-undefined benzaldehyde synthase and benzaldehyde dehydrogenase activities, respectively. The biosynthesis of biphenyls starts with benzoate-Coenzyme A ligase (BZL), which converts benzoic acid to benzoyl-CoA. Subsequently, the previously-defined biphenyl synthase uses benzoyl-CoA to form the biphenyls. The current study reports the first time detection and characterization of BZL activity in elicitor-treated pear cell cultures. The preferred substrate was benzoic acid (Km = 62 ± 4 µM). Magnesium or manganese was prerequisite for the activity, which was enhanced by ~ 70% in the presence of potassium. Maximum BZL activity was observed 18 h post elicitation, which is in agreement with the coordinate induction reported for the enzymes in the same pathway. The induced BZL activity preceded the accumulation of biphenyls supporting its involvement in their biosynthesis.


Assuntos
Compostos de Bifenilo/metabolismo , Coenzima A Ligases/genética , Células Vegetais , Pyrus/citologia , Sesquiterpenos/metabolismo , Acil Coenzima A/metabolismo , Benzaldeídos/metabolismo , Ácido Benzoico/metabolismo , Cinamatos/metabolismo , Coenzima A Ligases/metabolismo , Fenilalanina Amônia-Liase/metabolismo , Pyrus/metabolismo , Espectrometria de Massas em Tandem
15.
Talanta ; 206: 120195, 2020 Jan 01.
Artigo em Inglês | MEDLINE | ID: mdl-31514831

RESUMO

The ability to separate bioactive compounds from herbal medicines, which contain abundant components, is crucial for drug discovery. Conventional Countercurrent chromatography (CCC) methods for separating bioactive compounds are labor intensive and show low efficiency. Here, we present a novel integrative CCC method for separating lysine-specific demethylase 1 (LSD1) inhibitors from the roots of Salvia miltiorrhiza (RSM). The methanol extracts of RSM were separated into hydrosoluble and liposoluble fractions, which were online stored in coils. Subsequently, the targeting LSD1 constituents were isolated using isocratic, gradient, or recycling elution mode. All separation processes could be accomplished using one CCC apparatus. Using our separation strategy, two phenylpropanoids and four tanshinones were isolated, which were determined to be new classes of natural LSD1 inhibitors. Salvianolic acid B, which showed the most potent inhibitory activity with an IC50 of 0.11 µM, exhibiting a considerable potential as an anticancer agent. Promisingly, the integrative CCC could be a crucial tool for the target separation of enzyme inhibitors from herbal medicines.


Assuntos
Inibidores Enzimáticos/farmacologia , Histona Desmetilases/antagonistas & inibidores , Raízes de Plantas/química , Salvia miltiorrhiza/química , Benzofuranos/isolamento & purificação , Benzofuranos/metabolismo , Benzofuranos/farmacologia , Linhagem Celular Tumoral , Movimento Celular/efeitos dos fármacos , Cinamatos/isolamento & purificação , Cinamatos/metabolismo , Cinamatos/farmacologia , Distribuição Contracorrente/métodos , Depsídeos/isolamento & purificação , Depsídeos/metabolismo , Depsídeos/farmacologia , Inibidores Enzimáticos/isolamento & purificação , Inibidores Enzimáticos/metabolismo , Histona Desmetilases/metabolismo , Humanos , Simulação de Acoplamento Molecular , Fenantrenos/isolamento & purificação , Fenantrenos/metabolismo , Fenantrenos/farmacologia , Ligação Proteica
16.
PLoS One ; 14(12): e0226559, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-31841559

RESUMO

Iodine biofortification has been gaining interest in recent years as a sustainable and innovative approach to eradicate iodine deficiency disorders. Studying the impact of iodine biofortification on plant phenotype, biochemical and physiological parameters is crucial to leverage the expertise and best practices for the agro-food industry and human health. The aim of this study was to evaluate iodine biofortification on the main quantitative and qualitative traits of basil (Ocimum basilicum L.) plants cultivated both in open field and in growth chamber. The impact of KI and KIO3 treatments was evaluated on biomass production, as well as on the synthesis of phenolic compounds, especially rosmarinic acid and other caffeic acid derivatives, and on the essential oil (EO) composition. These compounds are typically accumulated in basil leaves and strongly contribute to the plant nutraceutical value and aroma. In open field, the use of increasing concentrations of both iodine salts gradually enhanced iodine accumulation in leaves, also determining an increase of the antioxidant power, total phenolics, rosmarinic acid and cinnamic acid accumulation. The composition of EO was only slightly affected by the treatments, as all the samples were characterized by a linalool chemotype and a minor alteration in their relative content was observed. A growth chamber experiment was performed to test EO variation in controlled conditions, broadening the range of iodine concentrations. In this case, plant chemotype was significantly affected by the treatments and large EO variability was observed, suggesting that iodine form and concentration can potentially influence the EO composition but that in open field this effect is overcome by environmental factors.


Assuntos
Biofortificação/métodos , Iodo/farmacologia , Ocimum basilicum/efeitos dos fármacos , Ocimum basilicum/metabolismo , Óleos Voláteis/metabolismo , Fenóis/metabolismo , Monoterpenos Acíclicos/análise , Monoterpenos Acíclicos/metabolismo , Agricultura/métodos , Biomassa , Cinamatos/análise , Cinamatos/metabolismo , Deficiências Nutricionais/prevenção & controle , Depsídeos/análise , Depsídeos/metabolismo , Ambiente Controlado , Humanos , Iodo/análise , Iodo/deficiência , Ocimum basilicum/química , Óleos Voláteis/análise , Fenóis/análise , Folhas de Planta/química , Folhas de Planta/efeitos dos fármacos , Folhas de Planta/metabolismo , Óleos Vegetais/análise , Óleos Vegetais/metabolismo
17.
Zhongguo Zhong Yao Za Zhi ; 44(21): 4704-4712, 2019 Nov.
Artigo em Chinês | MEDLINE | ID: mdl-31872668

RESUMO

Rosmarinic acid,a hydrosoluble polyphenolic hydroxyl compound,is the active ingredient in such traditional Chinese medicines as Menthae Haplocalycis Herba,Salviae Miltiorrhizae Radix et Rhizoma,Rosemary,Perillae Folium. Because of its good anti-inflammatory,anti-oxidant and anti-tumor effects,it is widely used in food,medicine and other fields. However,the metabolic process and metabolites of rosmarinic acid in vivo have not been completely defined. In this study,an efficient method of ultra-high performance liquid chromatography combined with linear ion trap-Orbitrap(UHPLC-LTQ-Orbitrap) mass spectrometer was used to analyze the metabolites in vivo of rosmarinic acid in rats. Plasma,urine and feces samples were collected after oral administration of rosmarinic acid. After biological samples were processed by solid phase extraction,Acquity UPLC  BEH C18 column(2. 1 mm × 100 mm,1. 7 µm) was used with 0. 1% formic acid(A)-acetonitrile(B) solution as the mobile phase at the speed of 0. 30 m L·min-1 and temperature of 35 ℃ under gradient conditions. The plasma,urine,feces and the blank samples were then analyzed by ESI-LTQ-Orbitrap under both negative and positive ion modes. Based on the accurate mass measurement(<5),MS/MS fragmentation patterns,standards and literatures,a total of 36 metabolites were screened out and identified in the biological samples collected from rats after intragastric administration. Three were identified 3 from rat plasma,31 from urine,and 7 from feces. The main metabolic pathways of rosmarinic acid in rats can be divided into five parts. Rosmarinic acid were first decomposed into small molecules,such as trans-caffeic acid,coumaric acid,m-hydroxybenzoic acid and Danshensu,which were followed by sulfation,methylation,glucuronic acid conjugation and glucose conjugation. The results showed that UHPLC-LTQ-Orbitrap mass spectrometer could be used to analyze the metabolism of rosmarinic acid in rats,and provide reference for further studies on toxicology,pharmacodynamics and secondary development of Chinese medicine.


Assuntos
Cinamatos/metabolismo , Depsídeos/metabolismo , Medicamentos de Ervas Chinesas/metabolismo , Espectrometria de Massas em Tandem , Animais , Cromatografia Líquida de Alta Pressão , Ratos
18.
Sci Rep ; 9(1): 17153, 2019 11 20.
Artigo em Inglês | MEDLINE | ID: mdl-31748605

RESUMO

Lignin is a complex phenylpropanoid polymer deposited in plant cell walls. Lignin has long been recognized as an important limiting factor for the polysaccharide-oriented biomass utilizations. To mitigate lignin-associated biomass recalcitrance, numerous mutants and transgenic plants that produce lignocellulose with reduced lignin contents and/or lignins with altered chemical structures have been produced and characterised. However, it is not fully understood how altered lignin chemistry affects the supramolecular structure of lignocellulose, and consequently, its utilization properties. Herein, we conducted comprehensive chemical and supramolecular structural analyses of lignocellulose produced by a rice cad2 mutant deficient in CINNAMYL ALCOHOL DEHYDROGENASE (CAD), which encodes a key enzyme in lignin biosynthesis. By using a solution-state two-dimensional NMR approach and complementary chemical methods, we elucidated the structural details of the altered lignins enriched with unusual hydroxycinnamaldehyde-derived substructures produced by the cad2 mutant. In parallel, polysaccharide assembly and the molecular mobility of lignocellulose were investigated by solid-state 13C MAS NMR, nuclear magnetic relaxation, X-ray diffraction, and Simon's staining analyses. Possible links between CAD-associated lignin modifications (in terms of total content and chemical structures) and changes to the lignocellulose supramolecular structure are discussed in the context of the improved biomass saccharification efficiency of the cad2 rice mutant.


Assuntos
Oxirredutases do Álcool/química , Oxirredutases do Álcool/metabolismo , Lignina/química , Lignina/metabolismo , Oryza/química , Oryza/metabolismo , Proteínas de Plantas/química , Biomassa , Parede Celular/química , Parede Celular/metabolismo , Cinamatos/química , Cinamatos/metabolismo , Estrutura Molecular , Mutação/genética , Proteínas de Plantas/metabolismo , Plantas Geneticamente Modificadas/química , Plantas Geneticamente Modificadas/metabolismo
19.
Bioorg Chem ; 93: 103316, 2019 12.
Artigo em Inglês | MEDLINE | ID: mdl-31585271

RESUMO

This study aimed to synthesize and screen tyrosinase inhibitors for delay fruit browning. A series of vanillyl cinnamate analogues were designed and synthesized by simple processes, and the inhibitory effects of all the synthesized derivatives on mushroom tyrosinase were evaluated. In the enzymatic activity test, compounds 21, 22, and 26 had significant (P < 0.05) effect on mushroom tyrosinase at a preliminary screening dose (1 mg/mL in vitro). IC50 analysis showed that the IC50 values of compounds 21, 22 and 26 were 268.5 µM, 213.2 µM and 413.5 µM, respectively. In the cytotoxicity evaluation, Cell Counting Kit-8 (CCK-8) assay showed that compounds 21, 22 and 26 had no significant effect on the proliferation of hepatocyte L02 and B16 melanoma cells at the dosage of 25-200 µM. Inhibition of tyrosinase activity and melanin content in B16 melanoma cells investigations indicated that compounds 21, 22 and 26 inhibited both cellular tyrosinase activity and melanin content dose-dependently and more strongly than the reference standard arbutin. The UV-visible spectra showed compound 22 inhibits the formation of dopamine quinone, further the molecular docking analysis of compound 22 with tyrosinase (PDB: 2Y9X) indicated that compound 22 interacted with the amino acid residues of tyrosinase. The results of anti-browning test showed that compounds 21, 22 and 26 had significant tyrosinase inhibition and anti-browning effects on fresh-cut apple slices at 4 °C in 48 h. Compound 22 could be used as novel tyrosinase inhibitor to delay fruit browning.


Assuntos
Cinamatos/metabolismo , Inibidores Enzimáticos/síntese química , Monofenol Mono-Oxigenase/antagonistas & inibidores , Agaricales/enzimologia , Animais , Sítios de Ligação , Linhagem Celular Tumoral , Proliferação de Células/efeitos dos fármacos , Sobrevivência Celular/efeitos dos fármacos , Cinamatos/química , Cinamatos/farmacologia , Inibidores Enzimáticos/metabolismo , Inibidores Enzimáticos/farmacologia , Cinética , Melaninas/antagonistas & inibidores , Melaninas/metabolismo , Camundongos , Simulação de Acoplamento Molecular , Monofenol Mono-Oxigenase/metabolismo , Estrutura Terciária de Proteína , Relação Estrutura-Atividade
20.
J Nat Prod ; 82(11): 3140-3149, 2019 11 22.
Artigo em Inglês | MEDLINE | ID: mdl-31622095

RESUMO

A flavonoid glycoside, quercitrin (1), and two phenylpropanoyl sucrose derivatives, vanicoside B (2) and lapathoside C (3), were isolated for the first time from the herb Persicaria dissitiflora. Vanicoside B (2) exhibited antiproliferative activity against a panel of cancer cell lines in triple-negative breast cancer (TNBC) MDA-MB-231 cells. The underlying mechanisms of the antitumor activity of 2 were investigated in TNBC cells. Upregulation of cyclin-dependent kinase 8 (CDK8) was observed in a claudin-low molecular subtype of TNBC cells. A molecular modeling study indicated that 2 showed a high affinity for CDK8. Further investigations revealed that 2 suppressed CDK8-mediated signaling pathways and the expression of epithelial-mesenchymal transition proteins and induced cell cycle arrest and apoptosis in MDA-MB-231 and HCC38 TNBC cells. Moreover, 2 inhibited tumor growth without overt toxicity in a nude mouse xenograft model implanted with MDA-MB-231 cells. Taken together, these findings demonstrate the significance of CDK8 activity in TNBC and suggest a potential use of 2 as a therapeutic candidate for the treatment of aggressive human triple-negative breast cancer.


Assuntos
Antineoplásicos Fitogênicos/química , Antineoplásicos Fitogênicos/farmacologia , Cinamatos/química , Cinamatos/farmacologia , Quinase 8 Dependente de Ciclina/efeitos dos fármacos , Polygonaceae/química , Neoplasias de Mama Triplo Negativas/tratamento farmacológico , Animais , Apoptose/efeitos dos fármacos , Pontos de Checagem do Ciclo Celular/efeitos dos fármacos , Linhagem Celular Tumoral , Cinamatos/metabolismo , Quinase 8 Dependente de Ciclina/metabolismo , Ensaios de Seleção de Medicamentos Antitumorais , Feminino , Humanos , Camundongos , Camundongos Nus , Estrutura Molecular , Neoplasias de Mama Triplo Negativas/metabolismo , Ensaio Tumoral de Célula-Tronco , Ensaios Antitumorais Modelo de Xenoenxerto
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