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1.
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
2.
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
3.
Enzyme Microb Technol ; 131: 109392, 2019 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-31615678

RESUMO

Rosmarinic acid (RA), as a hydroxycinnamic acid ester of caffeic acid (CA) and 3,4-dihydroxyphenyllactic acid (3,4-DHPL), is a phenylpropanoid-derived plant natural product and has diverse biological activities. This work acts as a modular platform for microbial production using a two-cofactor (ATP and CoA) regeneration system to product RA based on a cell-free biosynthetic approach. Optimal activity of the reaction system was pH 8 and 30 °C. Total turnover number for ATP and CoA was 820.60 ±â€¯28.60 and 444.50 ±â€¯9.65, respectively. Based on the first hour data, the RA productivity reached 320.04 mg L-1 h-1 (0.889 mM L-1 h-1).


Assuntos
Trifosfato de Adenosina/metabolismo , Anti-Inflamatórios não Esteroides/metabolismo , Cinamatos/metabolismo , Coenzima A/metabolismo , Depsídeos/metabolismo , Sistema Livre de Células , Concentração de Íons de Hidrogênio , Temperatura Ambiente
4.
Biomed Chromatogr ; 33(9): e4599, 2019 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-31108569

RESUMO

Rosmarinic acid (RA) is a caffeic acid derivative and one of the most abundant and bioactive constituents in Java tea (Orthosiphon stamineus), which has significant biological activities. However, relatively few studies have been conducted to describe this compound's metabolites in vivo. Therefore, an ultra-high-performance liquid chromatography coupled to quadrupole-time-of-flight tandem mass spectrometry (UHPLC-QTOF-MS/MS) analysis with a three-step data mining strategy was established for the metabolic profile of RA. Firstly, the exogenously sourced ions were filtered out by the MarkerView software and incorporated with Microsoft Office Excel software. Secondly, a novel modified mass detects filter strategy based on the predicted metabolites was developed for screening the target ions with narrow, well-defined mass detection ranges. Thirdly, the diagnostic product ions and neutral loss filtering strategy were applied for the rapid identification of the metabolites. Finally, a total of 16 metabolites were reasonably identified in urine, bile and feces, while metabolites were barely found in plasma. The metabolites of RA could also be distributed rapidly in liver and kidney. Glucuronidation, methylation and sulfation were the primary metabolic pathways of RA. The present findings might provide the theoretical basis for evaluating the biological activities of RA and its future application.


Assuntos
Cromatografia Líquida de Alta Pressão/métodos , Cinamatos/análise , Cinamatos/metabolismo , Mineração de Dados/métodos , Depsídeos/análise , Depsídeos/metabolismo , Orthosiphon/química , Espectrometria de Massas em Tandem/métodos , Animais , Cinamatos/química , Cinamatos/farmacocinética , Depsídeos/química , Depsídeos/farmacocinética , Masculino , Ratos , Ratos Sprague-Dawley , Distribuição Tecidual
5.
Int J Mol Sci ; 20(10)2019 May 15.
Artigo em Inglês | MEDLINE | ID: mdl-31096565

RESUMO

Many medicinal plant species are currently threatened in their natural habitats because of the growing demand for phytochemicals worldwide. A sustainable alternative for the production of bioactive plant compounds are plant biofactories based on cell cultures and organs. In addition, plant extracts from biofactories have significant advantages over those obtained from plants, since they are free of contamination by microorganisms, herbicides and pesticides, and they provide more stable levels of active ingredients. In this context, we report the establishment of Satureja khuzistanica cell cultures able to produce high amounts of rosmarinic acid (RA). The production of this phytopharmaceutical was increased when the cultures were elicited with coronatine and scaled up to a benchtop bioreactor. S. khuzistanica extracts enriched in RA were found to reduce the viability of cancer cell lines, increasing the sub-G0/G1 cell population and the activity of caspase-8 in MCF-7 cells, which suggest that S. khuzistanica extracts can induce apoptosis of MCF-7 cells through activation of the extrinsic pathway. In addition, our findings indicate that other compounds in S. khuzistanica extracts may act synergistically to potentiate the anticancer activity of RA.


Assuntos
Aziridinas/farmacologia , Cinamatos/metabolismo , Cinamatos/farmacologia , Cicloexenos/farmacologia , Depsídeos/metabolismo , Depsídeos/farmacologia , Espécies em Perigo de Extinção , Extratos Vegetais/farmacologia , Satureja/metabolismo , Adenocarcinoma/tratamento farmacológico , Reatores Biológicos , Caspase 8/metabolismo , Caspases/metabolismo , Técnicas de Cultura de Células , Pontos de Checagem do Ciclo Celular/efeitos dos fármacos , Linhagem Celular Tumoral/efeitos dos fármacos , Sobrevivência Celular/efeitos dos fármacos , Células Hep G2/efeitos dos fármacos , Humanos , Células MCF-7 , Compostos Fitoquímicos/farmacologia , Plantas Medicinais/química , Satureja/crescimento & desenvolvimento
6.
J Pharm Biomed Anal ; 172: 126-138, 2019 Aug 05.
Artigo em Inglês | MEDLINE | ID: mdl-31035094

RESUMO

Salvia miltiorrhiza Bunge is a traditional Chinese medicine, and its water-soluble phenolic acid active compounds have very important medicinal value; however, the synthesis pathways of the main active ingredients remain unknown. Here, we employed nuclear magnetic resonance (NMR)-based metabolomics and transcriptomics techniques to study the biosynthesis mechanism of salvianolic acids. High-performance liquid chromatography (HPLC) combined with NMR showed an improvement over traditional techniques, and 54 metabolites were detected. The results of the multivariate statistical analysis showed that salvianolic acid B (SAB), rosmarinic acid (RA), caffeic acid, succinate, and citrate were among the multiple compounds that were increased in the methyl jasmonate (MeJA)-elicited group; the levels of sucrose, fructose, glutamine, and tyrosine were decreased. Combined with the differentially expressed genes (DEGs) found by transcriptome sequencing, we speculate that the synthesis of RA after MeJA treatment mostly occurred through caffeic acid and bypassed 4-hydroxyphenyllactic acid. This provides useful information for the study of salvianolic acids synthesis.


Assuntos
Benzofuranos/metabolismo , Cinamatos/metabolismo , Depsídeos/metabolismo , Espectroscopia de Prótons por Ressonância Magnética/métodos , Salvia miltiorrhiza/metabolismo , Acetatos/farmacologia , Vias Biossintéticas/efeitos dos fármacos , Células Cultivadas , Cromatografia Líquida de Alta Pressão/métodos , Ciclopentanos/farmacologia , Perfilação da Expressão Gênica/métodos , Metabolômica/métodos , Oxilipinas/farmacologia , Fenilpropionatos/metabolismo , Raízes de Plantas/citologia
7.
Biomed Pharmacother ; 110: 111-117, 2019 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-30466000

RESUMO

In light of the widespread use of herbal medicines containing rosmarinic acid (RA) and limited literature available thereon, we investigated the metabolic interactions of RA with human cytochrome P450 monooxygenases (CYPs) and uridine diphosphate glucuronosyltransferases (UGTs). The involvement of selected enzymes (CYP1A2, CYP2C19, CYP2E1, CYP3 A4, UGT1A1, UGT1A6, and UGT2B7) in the metabolism of RA and the inhibitory effect of RA on the enzyme activity were comprehensively evaluated using human recombinant isozyme system. Additionally, concentration-dependent RA metabolism mediated by phase I enzymes (including CYPs) or UGT was investigated in human liver microsome (HLM) system. A significant disappearance of RA was observed in the seven CYP and UGT isoforms studied, indicating their possible involvement in the metabolism of RA. Based on Michaelis-Menten kinetics, the metabolism study using HLM suggests that the UGT system may have a higher capacity and lower affinity for the metabolism of RA than phase I enzyme (including CYP) systems. Moreover, RA weakly inhibited CYP2C9 and 2E1 activities with IC50 values of 39.6 and 61.0 µM, respectively, while moderately inhibiting UGT1A1, 1A6, and 2B7 with IC50 values of 9.24, 19.1, and 23.4 µM, respectively. By constructing Line weaver-Burk plots, the type of inhibition exhibited by RA on CYP and UGT activities was determined as follows: CYP2C19, mixed inhibition; CYP2E1, UGT1A1, UGT1A6, and UGT2B7, competitive inhibition. Based on the comparison of the IC50 and Ki values obtained in the current study with the previously reported plasma concentrations of RA after oral dosing in humans, it is suggested that RA may significantly inhibit the activities of the tested UGTs, rather than CYPs, in clinical settings. Thus, the present study could provide a basis for further studies on clinically significant interactions between UGT substrate drugs and herbal medicines containing RA.


Assuntos
Cinamatos/metabolismo , Inibidores das Enzimas do Citocromo P-450/metabolismo , Sistema Enzimático do Citocromo P-450/metabolismo , Depsídeos/metabolismo , Metabolismo Energético/fisiologia , Glucuronosiltransferase/antagonistas & inibidores , Glucuronosiltransferase/metabolismo , Anti-Inflamatórios não Esteroides/metabolismo , Anti-Inflamatórios não Esteroides/farmacologia , Cinamatos/farmacologia , Inibidores das Enzimas do Citocromo P-450/farmacologia , Depsídeos/farmacologia , Relação Dose-Resposta a Droga , Metabolismo Energético/efeitos dos fármacos , Humanos
8.
Int J Mol Sci ; 19(12)2018 Nov 28.
Artigo em Inglês | MEDLINE | ID: mdl-30487420

RESUMO

Jasmonic acid (JA) carboxyl methyltransferase (JMT), a key enzyme in jasmonate-regulated plant responses, may be involved in plant defense and development by methylating JA to MeJA, thus influencing the concentrations of MeJA in plant. In this study, we isolated the JMT gene from Salvia miltiorrhiza, an important medicinal plant widely used to treat cardiovascular disease. We present a genetic manipulation strategy to enhance the production of phenolic acids by overexpresion SmJMT in S. miltiorrhiza. Global transcriptomic analysis using RNA sequencing showed that the expression levels of genes involved in the biosynthesis pathway of phenolic acids and MeJA were upregulated in the overexpression lines. In addition, the levels of endogenous MeJA, and the accumulation of rosmarinic acid (RA) and salvianolic acid (Sal B), as well as the concentrations of total phenolics and total flavonoids in transgenic lines, were significantly elevated compared with the untransformed control. Our results demonstrate that overexpression of SmJMT promotes the production of phenolic acids through simultaneously activating genes encoding key enzymes involved in the biosynthesis pathway of phenolic acids and enhancing the endogenous MeJA levels in S. miltiorrhiza.


Assuntos
Hidroxibenzoatos/metabolismo , Metiltransferases/metabolismo , Proteínas de Plantas/metabolismo , Salvia miltiorrhiza/metabolismo , Cinamatos/metabolismo , Ciclopentanos/metabolismo , Depsídeos/metabolismo , Regulação da Expressão Gênica de Plantas/genética , Regulação da Expressão Gênica de Plantas/fisiologia , Metiltransferases/genética , Oxilipinas/metabolismo , Proteínas de Plantas/genética , Plantas Geneticamente Modificadas/genética , Plantas Geneticamente Modificadas/metabolismo , Salvia miltiorrhiza/genética
9.
J Food Sci ; 83(8): 2092-2100, 2018 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-30007045

RESUMO

The covalent interactions between whey protein isolate (WPI) and rosmarinic acid (RosA) at two different conditions, alkaline (pH 9) and enzymatic (in the presence of tyrosinase, PPO), at room temperature with free atmospheric air were studied. The conjugates formed between WPI and RosA were characterized in terms of their physicochemical and functional properties. The changes in protein structure were analyzed by intrinsic fluorescence and binding of 8-anilino-1-naphthalenesulfonic acid. The findings show that the covalent interactions caused a decrease in free amino and thiol groups and tryptophan content at both conditions. The decrease at enzymatic conditions was lower than at alkaline conditions. In addition, modified WPI at alkaline conditions exhibited higher antioxidative capacity compared to the modification at enzymatic conditions. However, WPI modified at enzymatic condition showed mild antimicrobial activity against Staphylococcus aureus LMG 10147 and MU50 compared to WPI modified at alkaline conditions and unmodified WPI (control). The modified WPI can be used as multifunctional ingredient into various food products with an additional health promoting effect of the bound phenolic compounds.


Assuntos
Antibacterianos , Antioxidantes , Cinamatos/química , Cinamatos/metabolismo , Depsídeos/química , Depsídeos/metabolismo , Proteínas do Soro do Leite/química , Proteínas do Soro do Leite/metabolismo , Fenômenos Químicos , Concentração de Íons de Hidrogênio , Monofenol Mono-Oxigenase/metabolismo , Fenóis
10.
Appl Microbiol Biotechnol ; 102(18): 7775-7793, 2018 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-30022261

RESUMO

Rosmarinic acid (RA) is a highly valued natural phenolic compound that is very commonly found in plants of the families Lamiaceae and Boraginaceae, including Coleus blumei, Heliotropium foertherianum, Rosmarinus officinalis, Perilla frutescens, and Salvia officinalis. RA is also found in other members of higher plant families and in some fern and horned liverwort species. The biosynthesis of RA is catalyzed by the enzymes phenylalanine ammonia lyase and cytochrome P450-dependent hydroxylase using the amino acids tyrosine and phenylalanine. Chemically, RA can be produced via methods involving the esterification of 3,4-dihydroxyphenyllactic acid and caffeic acid. Some of the derivatives of RA include melitric acid, salvianolic acid, lithospermic acid, and yunnaneic acid. In plants, RA is known to have growth-promoting and defensive roles. Studies have elucidated the varied pharmacological potential of RA and its derived molecules, including anticancer, antiangiogenic, anti-inflammatory, antioxidant, and antimicrobial activities. The demand for RA is therefore, very high in the pharmaceutical industry, but this demand cannot be met by plants alone because RA content in plant organs is very low. Further, many plants that synthesize RA are under threat and near extinction owing to biodiversity loss caused by unscientific harvesting, over-collection, environmental changes, and other inherent features. Moreover, the chemical synthesis of RA is complicated and expensive. Alternative approaches using biotechnological methodologies could overcome these problems. This review provides the state of the art information on the chemistry, sources, and biosynthetic pathways of RA, as well as its anticancer properties against different cancer types. Biotechnological methods are also discussed for producing RA using plant cell, tissue, and organ cultures and hairy-root cultures using flasks and bioreactors. The recent developments and applications of the functional genomics approach and heterologous production of RA in microbes are also highlighted. This chapter will be of benefit to readers aiming to design studies on RA and its applicability as an anticancer agent.


Assuntos
Antineoplásicos Fitogênicos/biossíntese , Cinamatos/metabolismo , Depsídeos/metabolismo , Neoplasias/tratamento farmacológico , Plantas/genética , Animais , Antineoplásicos Fitogênicos/farmacologia , Vias Biossintéticas , Biotecnologia , Cinamatos/farmacologia , Depsídeos/farmacologia , Genômica , Humanos , Plantas/metabolismo
11.
Chem Biodivers ; 15(10): e1800190, 2018 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-30027606

RESUMO

Horminum pyrenaicum L. is a medicinal, aromatic and ornamental plant native to the Pyrenees and Alps. The phytochemical composition is affected by various ecological factors, climatic conditions and cultivating factors, and especially phenological growth stages. Flavonoids, phenolic acids and triterpenic acids were identified and quantified in the above-ground parts of H. pyrenaicum during the phenological stages. The massive flowering was distinguished with the greatest total amounts of phenolic compounds (22232.8 µg/g), and rosmarinic acid was the predominant compound. The amounts of triterpenic compounds were at the lowest during the intense growth stage and significantly increased up to the massive flowering. The prevailing compounds were ursolic acid and betulinic acid, 12092.4 µg/g and 2618.9 µg/g, respectively. The major essential oil compounds were ß-phellandrene (56.6%), caryophyllene oxide (5.9%), (Z)-caryophyllene (4.4%), myrcene (4.1%) and thymol (3.6%). Selecting the optimal harvesting time could ensure the herbal raw material rich with preferred bioactive compounds.


Assuntos
Flavonoides/análise , Lamiaceae/química , Óleos Voláteis/química , Triterpenos/análise , Cinamatos/análise , Cinamatos/metabolismo , Depsídeos/análise , Depsídeos/metabolismo , Flavonoides/metabolismo , Hidroxibenzoatos/análise , Hidroxibenzoatos/metabolismo , Lamiaceae/crescimento & desenvolvimento , Óleos Voláteis/metabolismo , Triterpenos/metabolismo
12.
Environ Microbiol ; 20(12): 4230-4244, 2018 12.
Artigo em Inglês | MEDLINE | ID: mdl-30051572

RESUMO

The interference of plant compounds with bacterial quorum sensing (QS) is a major mechanism through which plants and bacteria communicate. However, little is known about the modes of action and effects on signal integrity during this type of communication. We have recently shown that the plant compound rosmarinic acid (RA) specifically binds to the Pseudomonas aeruginosa RhlR QS receptor. To determine the effect of RA on expression patterns, we carried out global RNA-seq analysis. The results show that RA induces the expression of 128 genes, amongst which many virulence factor genes. RA triggers a broad QS response because 88% of the induced genes are known to be controlled by QS, and because RA stimulated genes were found to be involved in all four QS signalling systems within P. aeruginosa. This finding was confirmed through the analysis of transcriptional fusions transferred to wt and a rhlI/lasI double mutant. RA did not induce gene expression in the rhlI/lasI/rhlR triple mutant indicating that the effects observed are due to the RA-RhlR interaction. Furthermore, RA induced seven sRNAs that were all encoded in regions close to QS and/or RA induced genes. This work significantly enhances our understanding of plant bacteria interaction.


Assuntos
Cinamatos/farmacologia , Depsídeos/farmacologia , Regulação Bacteriana da Expressão Gênica/efeitos dos fármacos , Pseudomonas aeruginosa/metabolismo , Percepção de Quorum/efeitos dos fármacos , Proteínas de Bactérias/genética , Proteínas de Bactérias/metabolismo , Proteínas de Transporte/metabolismo , Cinamatos/metabolismo , Depsídeos/metabolismo , Pseudomonas aeruginosa/efeitos dos fármacos , Percepção de Quorum/genética , Fatores de Virulência/genética , Fatores de Virulência/metabolismo
13.
Planta Med ; 84(12-13): 920-934, 2018 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-29843181

RESUMO

Lithospermum officinale is a valuable source of bioactive metabolites with medicinal and industrial values. However, little is known about genes involved in the biosynthesis of these metabolites, primarily due to the lack of genome or transcriptome resources. This study presents the first effort to establish and characterize de novo transcriptome assembly resource for L. officinale and expression analysis for three of its tissues, namely leaf, stem, and root. Using over 4Gbps of RNA-sequencing datasets, we obtained de novo transcriptome assembly of L. officinale, consisting of 77,047 unigenes with assembly N50 value as 1524 bps. Based on transcriptome annotation and functional classification, 52,766 unigenes were assigned with putative genes functions, gene ontology terms, and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathways. KEGG pathway and gene ontology enrichment analysis using highly expressed unigenes across three tissues and targeted metabolome analysis showed active secondary metabolic processes enriched specifically in the root of L. officinale. Using co-expression analysis, we also identified 20 and 48 unigenes representing different enzymes of lithospermic/chlorogenic acid and shikonin biosynthesis pathways, respectively. We further identified 15 candidate unigenes annotated as cytochrome P450 with the highest expression in the root of L. officinale as novel genes with a role in key biochemical reactions toward shikonin biosynthesis. Thus, through this study, we not only generated a high-quality genomic resource for L. officinale but also propose candidate genes to be involved in shikonin biosynthesis pathways for further functional characterization.


Assuntos
Benzofuranos/metabolismo , Ácido Clorogênico/metabolismo , Depsídeos/metabolismo , Lithospermum/genética , Metaboloma , Naftoquinonas/metabolismo , Transcriptoma , Vias Biossintéticas , Ontologia Genética , Lithospermum/química , Lithospermum/metabolismo , Folhas de Planta/química , Folhas de Planta/genética , Folhas de Planta/metabolismo , Raízes de Plantas/química , Raízes de Plantas/genética , Raízes de Plantas/metabolismo , Caules de Planta/química , Caules de Planta/genética , Caules de Planta/metabolismo
14.
Org Biomol Chem ; 16(16): 3038-3051, 2018 04 25.
Artigo em Inglês | MEDLINE | ID: mdl-29634062

RESUMO

Reinvestigating antibiotic scaffolds that were identified during the Golden Age of antibiotic discovery, but have long since been "forgotten", has proven to be an effective strategy for delivering next-generation antibiotics capable of combatting multidrug-resistant superbugs. In this study, we have revisited the trichloro-substituted depsidone, nidulin, as a selective and unexploited antibiotic lead produced by the fungus Aspergillus unguis. Manipulation of halide ion concentration proved to be a powerful tool for modulating secondary metabolite production and triggering quiescent pathways in A. unguis. Supplementation of the culture media with chloride resulted in a shift in co-metabolite profile to dichlorounguinols and nornidulin at the expense of the non-chlorinated parent, unguinol. Surprisingly, only marginal enhancement of nidulin was observed, suggesting O-methylation may be rate-limiting. Similarly, supplementation of the media with bromide led to the production of the corresponding bromo-analogues, but also resulted in a novel family of depsides, the unguidepsides. Unexpectedly, depletion of chloride from the media halted the biosynthesis of the non-chlorinated parent compound, unguinol, and redirected biosynthesis to a novel family of ring-opened analogues, the unguinolic acids. Supplementation of the media with a range of unnatural salicylic acids failed to yield the corresponding nidulin analogues, suggesting the compounds may be biosynthesised by a single polyketide synthase. In total, 12 new and 11 previously reported nidulin analogues were isolated, characterised and assayed for in vitro activity against a panel of bacteria, fungi and mammalian cells, providing a comprehensive structure-activity profile for the nidulin scaffold.


Assuntos
Antibacterianos/metabolismo , Aspergillus/metabolismo , Animais , Antibacterianos/química , Antibacterianos/farmacologia , Depsídeos/química , Depsídeos/metabolismo , Dibenzoxepinas/química , Dibenzoxepinas/metabolismo , Dibenzoxepinas/farmacologia , Resistência a Múltiplos Medicamentos/efeitos dos fármacos , Lactonas/química , Lactonas/metabolismo , Camundongos , Metabolismo Secundário , Relação Estrutura-Atividade
15.
Int J Mol Sci ; 19(2)2018 Feb 03.
Artigo em Inglês | MEDLINE | ID: mdl-29401682

RESUMO

Both caffeic acid and 3,4-dihydroxyphenyllactic acid (danshensu) are synthesized through two distinct routs of the shikimic acid biosynthesis pathway. In many plants, especially the rosemary and sage family of Lamiaceae, these two compounds are joined through an ester linkage to form rosmarinic acid (RA). A further structural diversity of RA derivatives in some plants such as Salvia miltiorrhiza Bunge is a form of RA dimer, salvianolic acid-B (SA-B), that further give rise to diverse salvianolic acid derivatives. This review provides a comprehensive perspective on the chemistry and pharmacology of these compounds related to their potential therapeutic applications to dementia. The two common causes of dementia, Alzheimer's disease (AD) and stroke, are employed to scrutinize the effects of these compounds in vitro and in animal models of dementia. Key pharmacological mechanisms beyond the common antioxidant and anti-inflammatory effects of polyphenols are highlighted with emphasis given to amyloid beta (Aß) pathologies among others and neuronal regeneration from stem cells.


Assuntos
Alcenos/farmacologia , Doença de Alzheimer/tratamento farmacológico , Cinamatos/farmacologia , Demência Vascular/tratamento farmacológico , Depsídeos/farmacologia , Fármacos Neuroprotetores/farmacologia , Nootrópicos/farmacologia , Polifenóis/farmacologia , Alcenos/isolamento & purificação , Alcenos/metabolismo , Doença de Alzheimer/genética , Doença de Alzheimer/metabolismo , Doença de Alzheimer/patologia , Peptídeos beta-Amiloides/antagonistas & inibidores , Peptídeos beta-Amiloides/genética , Peptídeos beta-Amiloides/metabolismo , Animais , Diferenciação Celular , Cinamatos/isolamento & purificação , Cinamatos/metabolismo , Demência Vascular/genética , Demência Vascular/metabolismo , Demência Vascular/patologia , Depsídeos/isolamento & purificação , Depsídeos/metabolismo , Modelos Animais de Doenças , Regulação da Expressão Gênica/efeitos dos fármacos , Humanos , Lactatos/metabolismo , Células-Tronco Neurais/efeitos dos fármacos , Células-Tronco Neurais/metabolismo , Células-Tronco Neurais/patologia , Neurônios/efeitos dos fármacos , Neurônios/metabolismo , Neurônios/patologia , Fármacos Neuroprotetores/isolamento & purificação , Fármacos Neuroprotetores/metabolismo , Nootrópicos/isolamento & purificação , Nootrópicos/metabolismo , Polifenóis/isolamento & purificação , Polifenóis/metabolismo , Acidente Vascular Cerebral/genética , Acidente Vascular Cerebral/metabolismo , Acidente Vascular Cerebral/patologia , Acidente Vascular Cerebral/prevenção & controle , Proteínas tau/antagonistas & inibidores , Proteínas tau/genética , Proteínas tau/metabolismo
16.
Molecules ; 23(2)2018 Jan 31.
Artigo em Inglês | MEDLINE | ID: mdl-29385035

RESUMO

Phytochemical characters were evaluated in a five-year-old lemon balm collection consisting of 15 and 13 subspecies officinalis and altissima accessions, respectively. Stems were lower in essential oil than leaves. First cut leaves (June) gave more oil than those of the second cut (August). Subspecies officinalis plants had leaf oils rich in geranial, neral and citronellal in various proportions in the first cut. However, in the second cut the oils from all accessions appeared very similar with 80-90% geranial plus neral. Leaf oils of subsp. altissima contained sesquiterpenes (ß-caryophyllene, caryophyllene oxide, germacrene D) and also further monoterpenes in the second cut. Leaves had higher rosmarinic acid (RA) contents than stems. More RA was in subsp. officinalis than subsp. altissima leaves. First cut leaves were richer in RA than those from second cut. Total phenolics and antioxidant parameters showed that lemon balm is a valuable source of plant antioxidants.


Assuntos
Aldeídos/química , Cinamatos/química , Depsídeos/química , Melissa/química , Monoterpenos/química , Óleos Voláteis/química , Folhas de Planta/química , Caules de Planta/química , Aldeídos/metabolismo , Cinamatos/metabolismo , Depsídeos/metabolismo , Melissa/metabolismo , Monoterpenos/metabolismo , Óleos Voláteis/metabolismo , Folhas de Planta/metabolismo , Caules de Planta/metabolismo
17.
Phytochemistry ; 148: 63-70, 2018 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-29421512

RESUMO

The CRISPR (Clustered Regularly Interspaced Short Palindromic Repeats)/Cas9 (CRISPR-associated) system is a powerful genome editing tool that has been used in many species. In this study, we focused on the phenolic acid metabolic pathway in the traditional Chinese medicinal herb Salvia miltiorrhiza, using the CRISPR/Cas9 system to edit the rosmarinic acid synthase gene (SmRAS) in the water-soluble phenolic acid biosynthetic pathway. The single guide RNA (sgRNA) was designed to precisely edit the most important SmRAS gene, which was selected from 11 family members through a bioinformatics analysis. The sequencing results showed that the genomes of 50% of the transgenic regenerated hairy roots had been successfully edited. Five biallelic mutants, two heterozygous mutants and one homozygous mutant were obtained from 16 independent transgenic hairy root lines when the sgRNA was driven by the Arabidopsis U6 promoter, while no mutants were obtained from 13 independent transgenic hairy root lines when the sgRNA was driven by the rice U3 promoter. Subsequently, expression and metabolomics analysis showed that the contents of phenolic acids, including rosmarinic acid (RA) and lithospermic acid B, and the RAS expression level were decreased in the successfully edited hairy root lines, particularly in the homozygous mutants. In addition, the level of the RA precursor 3,4-dihydroxyphenyllactic acid clearly increased. These results indicated that the CRISPR/Cas9 system can be utilized to identify important genes in a gene family with the assistance of bioinformatics analysis and that this new technology is an efficient and specific tool for genome editing in S. miltiorrhiza. This new system presents a promising potential method to regulate plant metabolic networks and improve the quality of traditional Chinese medicinal herbs.


Assuntos
Salvia miltiorrhiza/química , Benzofuranos/metabolismo , Sistemas CRISPR-Cas , Cinamatos/metabolismo , Depsídeos/metabolismo , Genoma de Planta , Hidroxibenzoatos/metabolismo , Lactatos/metabolismo , Mutagênese , Plantas Geneticamente Modificadas/genética , RNA Guia/metabolismo
18.
Redox Biol ; 15: 548-556, 2018 05.
Artigo em Inglês | MEDLINE | ID: mdl-29413966

RESUMO

Phenolic acids represent abundant components contained in human diet. However, the negative charge in their carboxylic group limits their capacity to diffuse through biological membranes, thus hindering their access to cell interior. In order to promote the diffusion of rosmarinic acid through biological membranes, we synthesized several lipophilic ester- and amide-derivatives of this compound and evaluated their capacity to prevent H2O2-induced DNA damage and apoptosis in cultured human cells. Esterification of the carboxylic moiety with lipophilic groups strongly enhanced the capacity of rosmarinic acid to protect cells. On the other hand, the amide-derivatives were somewhat less effective but exerted less cytotoxicity at high concentrations. Cell uptake experiments, using ultra-high performance liquid chromatography coupled with tandem mass spectrometry (UHPLC-MS/MS), illustrated different levels of intracellular accumulation among the ester- and amide-derivatives, with the first being more effectively accumulated, probably due to their extensive hydrolysis inside the cells. In conclusion, these results highlight the hitherto unrecognized fundamental importance of derivatization of diet-derived phenolic acids to unveil their biological potential.


Assuntos
Apoptose/efeitos dos fármacos , Cinamatos/farmacologia , Dano ao DNA/efeitos dos fármacos , Depsídeos/farmacologia , Estresse Oxidativo/efeitos dos fármacos , Amidas/química , Amidas/farmacologia , Cinamatos/química , Cinamatos/metabolismo , Citoplasma/efeitos dos fármacos , Citoplasma/metabolismo , Depsídeos/química , Depsídeos/metabolismo , Ésteres/química , Ésteres/farmacologia , Humanos , Peróxido de Hidrogênio/toxicidade , Ferro/química , Quelantes de Ferro/química , Células Jurkat , Espectrometria de Massas em Tandem
19.
J Photochem Photobiol B ; 179: 91-97, 2018 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-29351879

RESUMO

Shoot cultures of Aronia melanocarpa, A. arbutifolia and A. × prunifolia were maintained on Murashige and Skoog medium with 1 mg/l each of BA and NAA under monochromatic lights (far-red, red, blue lights, UV-A-irradiation), in darkness, and under white light (control). HPLC-DAD analyses of 19 phenolic acids and 11 flavonoids in methanolic extracts from the shoots revealed in all of them the presence of three depsides (chlorogenic, neochlorogenic and rosmarinic acids), protocatechuic acid, four flavonoid glycosides (cynaroside, quercitrin, hyperoside and rutoside), and additionally, in A. arbutifolia, 3,4-dihydroxyphenylacetic acid. Depending on light quality, the total amounts of these metabolites increased 1.8-5.9 times, reaching maximum values under blue light: 527.40 and 144.61 mg 100 g-1 DW (A. melanocarpa), 543.27 and 85.82 mg 100 g-1 DW (A. arbutifolia) and 1615.18 and 220.65 mg 100 g-1 DW (A. × prunifolia), respectively. The maximum total amounts were 1.3-3.6 times higher than under white light. The quantities of individual metabolites changed from 1.2 to 11.0 times, with high amounts of neochlorogenic acid and quercitrin in A. melanocarpa (243.35 and 75.64 mg 100 g-1 DW), and of chlorogenic and rosmarinic acids and quercitrin in A. arbutifolia (236.52, 219.35 and 51.01 mg 100 g-1 DW). Extremely high amounts of depsides (418.83, 644.68, 548.86 mg 100 g-1 DW) and quercitrin (165.88 mg 100 g-1 DW) were produced in cultures of the hybrid - A. × prunifolia. The results are potentially useful for practical applications. This is the first report documented the importance of light quality on the production of phenolic acids and flavonoids in three aronia in vitro cultures.


Assuntos
Flavonoides/metabolismo , Hidroxibenzoatos/metabolismo , Luz , Photinia/efeitos da radiação , Biomassa , Ácido Clorogênico/análise , Ácido Clorogênico/metabolismo , Cromatografia Líquida de Alta Pressão , Cinamatos/análise , Cinamatos/metabolismo , Depsídeos/análise , Depsídeos/metabolismo , Flavonoides/análise , Hidroxibenzoatos/análise , Photinia/química , Photinia/metabolismo , Brotos de Planta/química , Brotos de Planta/metabolismo , Brotos de Planta/efeitos da radiação , Quercetina/análogos & derivados , Quercetina/análise , Quercetina/metabolismo , Raios Ultravioleta
20.
Free Radic Biol Med ; 115: 232-245, 2018 02 01.
Artigo em Inglês | MEDLINE | ID: mdl-29221989

RESUMO

Phenolic acids have been associated to a wide range of important health benefits underlain by a common molecular mechanism of action. Considering that significant membrane permeation is prevented by their hydrophilic character, we hypothesize that their main effects result from the interplay with cell membrane surface. This hypothesis was tested using the paradigmatic caffeic acid (CA) and two of its ester derivatives, rosmarinic (RA) and chlorogenic (CGA) acids, for which we predict, based on molecular dynamics simulations, a shallow location in phospholipid bilayers dependent on the protonation-state. Using complementary experimental approaches, an interaction with the membrane was definitely revealed for the three compounds, with RA exhibiting the highest lipid bilayer partition, and the redox signals of membrane-bound RA and CA being clearly detected. Cholesterol decreased the compounds bilayer partition, but not their ability to lower membrane dipole potential. In more complex membrane models containing also sphingomyelin, with liquid disordered (ld)/ liquid ordered (lo) phases coexistence, mimicking domains in the external leaflet of human plasma membrane, all compounds were able to affect nanodomains lateral organization. RA, and to a lesser extent CGA, decreased the size of lo domains. The most significant effect of CA was the possible formation of a rigid gel-like phase, enriched in sphingomyelin. In addition, all phenolic acids decreased the order of lo domains. In sum, phenolic acid effects on the membrane are enhanced in cholesterol-rich lo phases, which predominate in the outer leaflet of human cell membranes and are involved in many key cellular processes.


Assuntos
Ácidos Cafeicos/química , Ácido Clorogênico/química , Cinamatos/química , Depsídeos/química , Hidroxibenzoatos/química , Bicamadas Lipídicas/química , Lipídeos de Membrana/química , Esfingomielinas/metabolismo , Ácidos Cafeicos/metabolismo , Membrana Celular/metabolismo , Células Cultivadas , Ácido Clorogênico/metabolismo , Colesterol/química , Colesterol/metabolismo , Cinamatos/metabolismo , Depsídeos/metabolismo , Ésteres/química , Humanos , Hidroxibenzoatos/metabolismo , Lipídeos de Membrana/metabolismo , Microdomínios da Membrana/metabolismo , Potenciais da Membrana , Simulação de Dinâmica Molecular , Fosfolipídeos , Esfingomielinas/química
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