RESUMEN
Abstract The antioxidant activity of Tetragonisca angustula honey (TAH) and its ethanolic extract (TAEE) were investigated. The total levels of phenolic (TPC) and flavonoids (TFC) were also evaluated. The results for TPC were 19.91 ± 0.38 and 29.37 ± 1.82 mg GAE g-1 and for TFC 0.20 ± 0.02 and 0.14 ± 0.01 mg QE g-1 of TAH and TAEE, respectively. Antioxidant activities were 73.29 ± 0.49% and 93.36 ± 0.27% in the DPPH● assay and 71.73 ± 4.07% and 97.86 ± 0.35% in ABTS●+ for TAH and TAEE, respectively. The total reducing activity was determined by the method of reducing power (PR) and iron ion (Fe III) and the results varied in PR from 151.7 ± 25.7 and 230.7 ± 25.2 mg GAE L-1, for TAH and TAEE respectively and for (Fe III) in EC50 0.284 in TAEE and 0.687 in TAH. Chemical analysis by HPLC-DAD of the ethanolic extract (TAEE) revealed the presence of ferulic acid as majority phenolic component in the extract. The 1H NMR analysis confirmed this structure and showed the also presence of glucose, citric acid, succinic acid, proline and hydrocarbon derivatives. In addition, the botanical origin was also investigated and showed a multifloral characteristic, having found 19 pollen types with a botanical predominance of the Anacardiaceae family, with Tapirira pollen occurring as predominant (42.6%) and Schinus as secondary (25.7%). The results showed that T. angustula honey is an interesting source of antioxidant phenolic compounds due to its floral origin and can act as a protector of human health when consumed.
Resumo A atividade antioxidante do mel de Tetragonisca angustula (TAH) e seu extrato etanólico (TAEE) foram investigados. Os níveis totais de fenólicos (TPC) e flavonóides (TFC) também foram avaliados. Os resultados para TPC foram 19,91 ± 0,38 e 29,37 ± 1,82 mg GAE g-1 e para TFC 0,20 ± 0,02 e 0,14 ± 0,01 mg QE g-1 de TAH e TAEE, respectivamente. As atividades antioxidantes foram 73,29 ± 0,49% e 93,36 ± 0,27% no ensaio DPPH● e 71,73 ± 4,07% e 97,86 ± 0,35% no ABTS●+ para TAH e TAEE, respectivamente. A atividade redutora total foi determinada pelo método de poder redutor (PR) e íon ferrico (Fe III) e os resultados variaram em PR de 151,7 ± 25,7 e 230,7 ± 25,2 mg GAE L-1, para TAH e TAEE respectivamente e para (Fe III) em EC50 0,284 em TAEE e 0,687 em TAH. A análise química por HPLC-DAD do extrato etanólico (TAEE) revelou a presença de ácido ferúlico como componente majoritário no extrato. A análise de RMN 1H confirmou esta estrutura e mostrou a presença de glicose, ácido cítrico, ácido succínico, prolina e derivados de hidrocarbonetos no TAEE. Além disso, a origem botânica também foi investigada e apresentou característica multifloral, tendo encontrado 19 tipos polínicos com predomínio botânico da família Anacardiaceae, sendo o pólen Tapirira predominante (42,6%) e o Schinus secundário (25,7%). Os resultados mostraram que o mel de T. angustula é uma interessante fonte de compostos fenólicos antioxidantes devido à sua origem floral e pode atuar como protetor da saúde humana quando consumido.
Asunto(s)
Humanos , Animales , Miel/análisis , Antioxidantes , Fenoles/análisis , Brasil , Ácidos CumáricosRESUMEN
OBJECTIVE@#To investigate the effect of ferulic acid, a natural compound, on pancreatic beta cell viability, Ca2+ channels, and insulin secretion.@*METHODS@#We studied the effects of ferulic acid on rat insulinoma cell line viability using the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide viability assay. The whole-cell patch-clamp technique and enzyme-linked immunosorbent assay were also used to examine the action of ferulic acid on Ca2+ channels and insulin secretion, respectively.@*RESULTS@#Ferulic acid did not affect cell viability during exposures up to 72 h. The electrophysiological study demonstrated that ferulic acid rapidly and concentration-dependently increased L-type Ca2+ channel current, shifting its activation curve in the hyperpolarizing direction with a decreased slope factor, while the voltage dependence of inactivation was not affected. On the other hand, ferulic acid have no effect on T-type Ca2+ channels. Furthermore, ferulic acid significantly increased insulin secretion, an effect inhibited by nifedipine and Ca2+-free extracellular fluid, confirming that ferulic acid-induced insulin secretion in these cells was mediated by augmenting Ca2+ influx through L-type Ca2+ channel. Our data also suggest that this may be a direct, nongenomic action.@*CONCLUSION@#This is the first electrophysiological demonstration that acute ferulic acid treatment could increase L-type Ca2+ channel current in pancreatic β cells by enhancing its voltage dependence of activation, leading to insulin secretion.
Asunto(s)
Ratas , Animales , Secreción de Insulina , Insulina/farmacología , Células Secretoras de Insulina/metabolismo , Ácidos Cumáricos/metabolismo , Calcio/metabolismoRESUMEN
p-coumaric acid is one of the aromatic compounds that are widely used in food, cosmetics and medicine due to its properties of antibacterium, antioxidation and cardiovascular disease prevention. Tyrosine ammonia-lyase (TAL) catalyzes the deamination of tyrosine to p-coumaric acid. However, the lack of highly active and specific tyrosine ammonia lyase limits cost-effective microbial production of p-coumaric acid. In order to improve biosynthesis efficiency of p-coumaric acid, two tyrosine ammonia-lyases, namely Fc-TAL2 derived from Flavobacterium columnare and Fs-TAL derived from Flavobacterium suncheonense, were selected and characterized. The optimum temperature (55 ℃) and pH (9.5) for Fs-TAL and Fc-TAL2 are the same. Under optimal conditions, the specific enzyme activity of Fs-TAL and Fc-TAL2 were 82.47 U/mg and 13.27 U/mg, respectively. Structural simulation and alignment analysis showed that the orientation of the phenolic hydroxyl group of the conserved Y50 residue on the inner lid loop and its distance to the substrate were the main reasons accounting for the higher activity of Fs-TAL than that of Fc-TAL2. The higher activity and specificity of Fs-TAL were further confirmed via whole-cell catalysis using recombinant Escherichia coli, which could convert 10 g/L tyrosine into 6.2 g/L p-coumaric acid with a yield of 67.9%. This study provides alternative tyrosine ammonia-lyases and may facilitate the microbial production of p-coumaric acid and its derivatives.
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Amoníaco-Liasas/química , Ácidos Cumáricos , Escherichia coli/genética , TirosinaRESUMEN
Objective: To investigate the regulatory effects and signaling mechanism of sodium ferulate on the proliferation and apoptosis of human skin hypertrophic scar fibroblasts (HSFbs). Methods: The experimental research methods were used. The 4th-6th passage of HSFbs from human skin were used for the following experiments. HSFbs were co-cultured with sodium ferulate at final mass concentrations of 1, 1×10-1, 1×10-2, 1×10-3, 1×10-4, 1×10-5, and 1×10-6 mg/mL for 48 hours, and methyl thiazolyl tetrazolium method was used to determine the cell absorbance values and linear regression was used to analyze the half lethal concentration (LC50) of sodium ferulate (n=6). HSFbs were co-cultured with sodium ferulate at final mass concentrations of 0.1, 0.2, 0.3, and 0.4 mg/mL for 24, 48, 72, and 96 hours, and methyl thiazolyl tetrazolium method was used to determine the cell absorbance values and the cell proliferation inhibition rate was calculated (n=3). According to the random number table, the cells were divided into 0.300 mg/mL sodium ferulate group, 0.030 mg/mL sodium ferulate group, 0.003 mg/mL sodium ferulate group treated with sodium ferulate at corresponding final mass concentrations, and negative control group without any treatment. After 72 hours of culture, the cell absorbance values were determined by methyl thiazolyl tetrazolium method (n=5), the microscopic morphology of cells was observed by transmission electron microscope (n=3), the cell apoptosis was detected by TdT-mediated dUTP-biotin nick end labeling (TUNEL) assay and the apoptosis index was calculated (n=4), the protein expressions of B lymphocystoma-2 (Bcl-2), Bcl-2-associated X protein (Bax), and cysteine aspartic acid specific protease-3 (caspase-3) were determined by immunohistochemistry (n=4), and the protein expressions of transformed growth factor β1 (TGF-β1), phosphorylated Smad2/3, phosphorylated Smad4, and phosphorylated Smad7 were detected by Western blotting (n=4). Data were statistically analyzed with one-way analysis of variance and Dunnett test. Results: The LC50 of sodium ferulate was 0.307 5 mg/mL. After being cultured for 24-96 hours, the cell proliferation inhibition rates of cells treated with sodium ferulate at four different mass concentrations tended to increase at first but decrease later, which reached the highest after 72 hours of culture, so 72 hours was chosen as the processing time for the subsequent experiments. After 72 hours of culture, the cell absorbance values in 0.003 mg/mL sodium ferulate group, 0.030 mg/mL sodium ferulate group, and 0.300 mg/mL sodium ferulate group were 0.57±0.06, 0.53±0.04, 0.45±0.05, respectively, which were significantly lower than 0.69±0.06 in negative control group (P<0.01). After 72 hours of culture, compared with those in negative control group, the cells in the three groups treated with sodium ferulate showed varying degrees of nuclear pyknosis, fracture, or lysis, and chromatin loss. In the cytoplasm, mitochondria were swollen, the rough endoplasmic reticulum was expanded, and local vacuolation gradually appeared. After 72 hours of culture, compared with that in negative control group, the apoptosis indexes of cells were increased significantly in 0.003 mg/mL sodium ferulate group, 0.030 mg/mL sodium ferulate group, and 0.300 mg/mL sodium ferulate group (P<0.05 or P<0.01). After 72 hours of culture, compared with those in negative control group, the protein expressions of Bcl-2 of cells in 0.300 mg/mL sodium ferulate group was significantly decreased (P<0.01), the protein expressions of Bax of cells in 0.030 mg/mL sodium ferulate group and 0.300 mg/mL sodium ferulate group were significantly increased (P<0.05), and the protein expression of caspase-3 of cells in 0.300 mg/mL sodium ferulate group was significantly increased (P<0.01). After 72 hours of culture, compared with those in negative control group, the protein expression levels of TGF-β1, phosphorylated Smad2/3, and phosphorylated Smad4 of cells in 0.030 mg/mL sodium ferulate group and 0.300 mg/mL sodium ferulate group were significantly decreased (P<0.05 or P<0.01), and the protein expression levels of phosphorylated Smad7 of cells in 0.003 mg/mL sodium ferulate group, 0.030 mg/mL sodium ferulate group, and 0.300 mg/mL sodium ferulate group were significantly increased (P<0.01). Conclusions: Sodium ferulate can inhibit the proliferation of HSFbs of human skin and promote the apoptosis of HSFbs of human skin by blocking the expression of key proteins on the TGF-β/Smad signaling pathway and synergistically activating the mitochon- drial apoptosis pathway.
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Humanos , Apoptosis , Caspasa 3/metabolismo , Proliferación Celular , Cicatriz Hipertrófica/metabolismo , Ácidos Cumáricos , Fibroblastos/metabolismo , Transducción de Señal , Proteína X Asociada a bcl-2/farmacologíaRESUMEN
O objetivo do tratamento endodôntico é manter a integridade da raiz, bem como, prevenir ou resolver doenças periapicais, pela erradicação dos microrganismos e de suas fontes de nutrientes provenientes do sistema de canais radiculares. A complexidade da anatomia dos canais radiculares e dos biofilmes multiespécies aumenta a dificuldade em eliminar os microrganismos e controlar a inflamação por procedimentos químico-mecânicos convencionais, o que justifica o uso de medicações intracanais. Novos compostos com amplo efeito antimicrobiano e potencial antiinflamatório, como os ácidos fenólicos, poderiam ser explorados como princípios ativos de medicamentos intracanais. Entretanto, para aumentar a solubilidade, controlar a liberação e estender os efeitos biológicos dos ácidos fenólicos, seria interessante incorporá-los em carreadores de medicamentos como os hidrogéis de quitosana. Este estudo foi dividido em dois capítulos que apresentaram como objetivos: 1) avaliar as atividades antimicrobiana, antibiofilme e antiinflamatória e a citotoxicidade do ácido cinâmico e seus derivados; 2) sintetizar e caracterizar as propriedades químicas e físico-mecânicas de hidrogéis termossensíveis de quitosana e poloxamer contendo ácidos fenólicos, e avaliar o efeito desses hidrogéis sobre biofilmes multiespécies e na viabilidade de macrófagos e fibroblastos. No capítulo 1, a atividade antimicrobiana do ácido cinâmico (CI) e seus derivados ácido cumárico (CO), ácido cafeico (CA), ácido ferúlico (FE) e ácido sinápico (SI) foi avaliada pela determinação da concentração inibitória e bactericida mínima (CIM/CBM) e Concentração Inibitória Fracionada (CIF) sobre Enterococcus faecalis, Streptococcus mutans, Lactobacillus casei, Actinomyces israelii e Fusobacterium nucleatum. Os ácidos fenólicos foram selecionados e seu efeito em biofilmes dual-espécies e multiespécies com as mesmas cepas padrão ou cepas clínicas foram avaliados por contagem bacteriana, microscopia eletrônica de varredura e microscopia confocal. A viabilidade de fibroblastos L929 e macrófagos RAW 264.7 na presença desses ácidos fenólicos foi avaliada por ensaios de resazurina. Além disso, os níveis de mRNA dos marcadores pró-inflamatórios TNF-α, IL-1ß, iNOS e COX-2 foram determinados por PCR quantitativo TaqMan após exposição de macrófagos aos ácidos fenólicos e ao lipopolissacarídeo (LPS). No capítulo 2, foi realizada a síntese e caracterização físicomecânica de hidrogéis de quitosana-poloxamer (CPH) contendo ácidos fenólicos e avaliado seus efeitos no biofilme multiespécies e na viabilidade de macrófagos e fibroblastos. Os dados foram analisados estatisticamente considerando p< 0,05. O ácido cinâmico e o ácido cafeico apresentaram efeito inibitório e bactericida contra todas as espécies bacterianas testadas, com os menores valores de CIM e CBM. Entretanto, não houve efeito sinérgico entre eles (FICI> 0,5). Ambos os compostos (5x a CIM mais alta) foram eficazes na eliminação de biofilmes dual-espécies e na redução significativa de biofilmes multiespécies, especialmente o ácido cinâmico. O ácido cinâmico causou toxicidade mínima para ambas as culturas celulares nas concentrações de CIM e o ácido cafeico não foi citotóxico em concentrações abaixo de 0,125 mg/mL. Ambos os compostos reduziram significativamente TNF-α, IL-1ß, iNOS e COX-2, de maneira dose-dependente. Os CPH foram caracterizados como termorreversíveis e com propriedades mecânicas e bioadesivas desejáveis. O efeito dos hidrogéis CPH+CA (77,8%) e CPH+CI (73,2%) em reduzir os biofilmes multiespécies foi superior ao CPH+ hidroxido de cálcio (CH) (53,6%) e CPH+ clorexidina (CHX) (39,9%). Em geral, CPH + CI causou menor citotoxicidade quando comparado a CPH + CA, para ambas as linhagens celulares. Conclui-se que o ácido cinâmico e ácido cafeico apresentaram efeito bactericida e contra biofilmes formados por bactérias associadas com infecções endodônticas, causando baixa citotoxicidade. Ambos os compostos apresentaram efeito antiinflamatório, inibindo a expressão de marcadores próinflamatórios em macrófagos estimulados por LPS. Os hidrogéis de quitosana-poloxamer foram termorreversíveis e apresentaram adequadas propriedades mecânicas e adesivas para aplicação clínica, e quando combinados principalmente com ácido cinâmico, promoveram a redução de biofilmes multiespécies formados nos túbulos dentinários, causando baixa toxicidade em fibroblastos e macrófagos(AU)
The objective of endodontic treatment is to maintain the integrity of the root, as well as to prevent or resolve periapical diseases, by eradicating microorganisms and their sources of nutrients from the root canal system. The complexity of root canal anatomy and multispecies biofilms increases the difficulty in eliminating microorganisms and controlling inflammation by conventional chemical-mechanical procedures, which justifies the use of intracanal medications. New compounds with broad antimicrobial effect and anti-inflammatory potential, such as phenolic acids, could be explored as active principles of intracanal medications. However, to increase the solubility, control the release and extend the biological effects of phenolic acids, it would be interesting to incorporate them into drug carriers such as chitosan hydrogels. This study was divided into two chapters with the following objectives: 1) to evaluate the antimicrobial, antibiofilm and anti-inflammatory activities and the cytotoxicity of cinnamic acid and its derivatives; 2) synthesize and characterize chemical and physicomecanical properties of thermosensitive chitosan and poloxamer hydrogels containing phenolic acids and evaluate the effect of these hydrogels on multispecies biofilms and on the viability of macrophages and fibroblasts. In chapter 1, the antimicrobial activity of cinnamic acid (CI) and its derivatives coumaric acid (CO), caffeic acid (CA), ferulic acid (FE) and sinapic acid (SI) was evaluated by determining the minimum inhibitory and bactericidal concentration (MIC/MBC) and Fractional Inhibitory Concentration (FIC) on Enterococcus faecalis, Streptococcus mutans, Lactobacillus casei, Actinomyces israelii and Fusobacterium nucleatum. Phenolic acids were selected and their effect on bispecies and multispecies biofilms with the same standard or clinical strains were evaluated by bacterial counts, scanning electron microscopy and confocal microscopy. The viability of L929 fibroblasts and RAW 264.7 macrophages in the presence of these phenolic acids was evaluated by resazurin assays. In addition, mRNA levels of the proinflammatory markers TNF-α, IL-1ß, iNOS and COX-2 were determined by quantitative TaqMan PCR after macrophage exposure to phenolic acids and lipopolysaccharide (LPS). In chapter 2, the synthesis and physical-mechanical characterization of chitosanpoloxamer (CPH) hydrogels containing phenolic acids were performed and their effects on multispecies biofilm and on the viability of macrophages and fibroblasts were evaluated. Data were statistically analyzed considering p< 0.05. Cinnamic acid and caffeic acid showed an inhibitory and bactericidal effect against all bacterial species tested, with the lowest MIC and MBC values. However, no synergistic effect was observed between the compounds (FICI> 0.5). Both compounds (at 5x the highest MIC) were effective in eliminating dual-species biofilms and significantly decreasing multispecies biofilms, especially cinnamic acid. Cinnamic acid caused minimal toxicity to both cell cultures at MIC concentrations and caffeic acid was not cytotoxic at concentrations below 0.125 mg/mL. Both compounds significantly reduced TNF-α, IL1ß, iNOS and COX-2, in a dose-dependent manner. CPH were characterized as thermoreversible and with adequate mechanical and bioadhesive properties. The effect of CPH+CA (77.8%) and CPH+CI (73.2%) hydrogels against multispecies biofilms was superior to CPH + calcium hydroxide (CH) (53.6%) and CPH + chlorhexidine (CHX) (39.9%). In general, CPH + CI caused less cytotoxicity when compared to CPH + CA, for both cell lines. In conclusion, cinnamic acid and caffeic acid showed bactericidal effect and against biofilms of bacteria associated with endodontic infections, causing minimal cytotoxicity. In addition, both compounds showed an anti-inflammatory effect, inhibiting the expression of pro-inflammatory markers in LPS-stimulated macrophages. The chitosan-poloxamer hydrogels were thermoreversible and presented adequate mechanical and bioadhesive properties for clinical application, and when combined specially with cinnamic acid, they promoted the reduction of multispecies biofilms formed in the dentinal tubules, causing low toxicity to fibroblasts and macrophages(AU)
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Ácidos Cafeicos , Cinamatos , Cinamatos/toxicidad , Streptococcus mutans , Actinomyces , Fusobacterium nucleatum , Enterococcus faecalis , Poloxámero , Ácidos Cumáricos , Compuestos Fenólicos , Lacticaseibacillus casei , AntiinfecciososRESUMEN
ABSTRACT Purpose To investigate the effect of ferulic acid (FA) on spinal cord injury (SCI)-induced motor dysfunction and to explore the possible pharmacological mechanisms. Methods Adult male Wistar rats were used in our study. SCI was achieved by clipping the spinal cord T9 of the rat by a vascular clip for 2 minutes. The motor function of the rat was evaluated by Basso, Beattie, and Bresnahan scoring method (BBB) and inclined plane test. Hematoxylin and eosin (HE) staining, NISSL staining, and transmission electron microscopic examination were used to evaluate alterations at the histological level. Polymerase chain reaction (PCR), Western blots, and enzyme-linked immunosorbent assays (ELISA) were employed in biochemical analysis. Results The BBB score and inclined plane test score significantly decreased after SCI surgery, whereas chronic FA treatment (dose of 90 mg/kg, i.g.) for 28 days improved SCI-induced motor dysfunction. HE staining showed that SCI surgery induced internal spinal cord edema, but the structural changes of the spinal cord could be reversed by FA treatment. NISSL staining and transmission electron microscopic examination confirmed the improvement of the effect of FA on the injury site. In the biochemical analysis, it could be found that FA inhibitedSCI-induced mRNA and protein overexpression of pro-inflammatory cytokines (IL-1β, IL-6, TNF-α), as well as iNOS and COX-2 via the modulation of NF-κB level in the spinal cord of SCI rat. Moreover, the SCI-induced decrease of Bcl-2/Bax ratio was also reversed by FA treatment. However, the effect of FA on the expression of Beclin-1 was not statistically significant. Conclusions FA showed a therapeutic effect on SCI, which may be associated with the regulation of neuroinflammation and apoptosis.
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Animales , Masculino , Ratas , Traumatismos de la Médula Espinal/tratamiento farmacológico , Médula Espinal , Ratas Wistar , Ratas Sprague-Dawley , Apoptosis , Ácidos Cumáricos , Recuperación de la FunciónRESUMEN
Ischemia-reperfusion injury (IRI) has brought attention to flap failure in reconstructive surgery. To improve the prognosis of skin transplantation, we performed experimental IRI by surgical obstruction of blood flow and used sodium ferulate (SF) to prevent IRI in rats. After SF treatment, the morphological and histological changes of the skin flaps were observed by H&E and Masson's trichrome staining. We also detected the expression levels of COX-1, HO-1, and Ki67 by immunohistochemical and western blot analysis. Moreover, enzyme-linked immunosorbent assay was used to identify the content of tumor necrosis factor (TNF)-α, myeloperoxidase (MPO), malondialdehyde (MDA), and nitric oxide (NO) in peripheral blood and skin tissue. Compared with the model group, SF treatment significantly improved the recovered flap area (%) and promoted collagen synthesis. Cyclooxygenase-2 (COX-2) expression was significantly inhibited by heme oxygenase-1 (HO-1) induction after SF treatment. Furthermore, SF significantly inhibited the levels of TNF-α in peripheral blood, MPO and MDA in the skin tissue, and the increased synthesis of NO. Our results showed the protective effects of SF on IRI after flap transplantation and we believe that the protective effects of SF was closely related to the alleviation of the inflammatory response and the inhibition of the oxidative stress injury.
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Animales , Ratas , Daño por Reperfusión/prevención & control , Daño por Reperfusión/tratamiento farmacológico , Estrés Oxidativo , Ácidos Cumáricos/farmacología , Antiinflamatorios/farmacologíaRESUMEN
Plants of the genera Werneria (Asteraceae) and Xenophyllum (genus extracted from Werneria) are used in traditional medicine of Latin America for the treatment of mountain sickness, hypertension and gastrointestinal disorders. Only a small number of species of these genera have been studied, leading to the isolation of compounds belonging to the classes of benzofurans, chromenes, acetophenones, coumarates, diterpenes and pyrrolizidine alkaloids. Some of the plant extracts and/or compounds have shown antimicrobial, anti-HIV, hypotensive and photoprotective activities.
Las plantas de los geÌneros Werneria (Asteraceae) y Xenophyllum (geÌnero extraido de Werneria) son usadas en la medicina tradicional de AmeÌrica Latina para el tratamiento del mal de montanÌa, hipertensioÌn y desoÌrdenes gastrointestinales. Solo un pequenÌo nuÌmero de especies de estos geÌneros ha sido investigado, lograÌndose aislar compuestos que pertenecen a las clases de benzofuranos, cromenos, acetofenonas, cumaratos, diterpenos y alcaloides pirrolizidiÌnicos. Algunos de los extractos y/o compuestos de dichas plantas han mostrado actividades antimicrobianas, anti-HIV, hipotensoras y fotoprotectoras.
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Plantas Medicinales/química , Extractos Vegetales/uso terapéutico , Asteraceae/química , Acetofenonas/química , Terpenos/análisis , Benzopiranos/química , Flavonoides/química , Ácido Clorogénico/química , Ácidos Cumáricos/química , Alcaloides/química , Mal de Altura/tratamiento farmacológico , Hipertensión/tratamiento farmacológico , Medicina TradicionalRESUMEN
Caffeic acid (CA; 3,4-dihydroxycinnamic acid) is an aromatic compound obtained by the phenylpropanoid pathway. This natural product has antioxidant, antitumor, antiviral, and anti-inflammatory activities. It is also a precursor of CA phenethyl ester (CAPE), a compound with potential as an antidiabetic and liver-protective agent. CA can be found at low concentrations in plant tissues, and hence, its purification is difficult and expensive. Knowledge regarding the pathways, enzymes, and genes involved in CA biosynthesis has paved the way for enabling the design and construction of microbial strains with the capacity of synthesizing this metabolite. In this review, metabolic engineering strategies for the generation of Escherichia coli strains for the biotechnological production of CA are presented and discussed.
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Ácidos Cafeicos/metabolismo , Escherichia coli/metabolismo , Ingeniería Metabólica/métodos , Productos Biológicos , Biotecnología , Ácidos CumáricosRESUMEN
Ten batches of Angelica sinensis from three producing areas( Tuoxiang,Minxian and Weiyuan of Gansu province) were selected as the research objects,and processed into raw A. sinensis,A. sinensis with alcohol,and A. sinensis with soil respectively through the standard processing methods. Ultra-high performance liquid chromatography( UPLC) was used to establish fingerprint for three processed products of A. sinensis,and determine the contents of 9 phenolic acids and phthalide compounds. The similarity was analyzed with Similarity Evaluation System for Chromatographic Fingerprint of Traditional Chinese Medicine,which showed that the chromatographic peaks of the same processed samples of A. sinensis were basically similar,with all similarities greater than 0. 950. The difference between different processed products and their control spectra was not obvious,with all similarities also higher than 0. 950.On the basis of using principal component analysis( PCA) and OPLS-DA to seek the difference components between groups,the improved distance coefficient method can be used to effectively distinguish the three processed products of A. sinensis by fingerprint similarity. At the same time,the determination method of nine phenolic acids and phthalide in A. sinensis was established by UPLC,and the comparison between different processed products was carried out. The results showed that the content of various components was changed as compared with the raw A. sinensis. The contents of coniferyl ferulate and ligustilide in the A. sinensis with alcohol were increased significantly,and the content of coniferyl ferulate was obviously increased in A. sinensis with soil. The method established in this paper can effectively distinguish different processed products of A. sinensis and determine the content of the main components in them.
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4-Butirolactona , Angelica sinensis , Química , Benzofuranos , Cromatografía Líquida de Alta Presión , Ácidos Cumáricos , Medicamentos Herbarios Chinos , Hidroxibenzoatos , Medicina Tradicional China , Análisis de Componente PrincipalRESUMEN
OBJECTIVE@#A strain of Aspergillus niger (A. niger), capable of releasing bound phenolic acids from wheat bran, was isolated. This strain was identified by gene sequence identification. The antioxidant and anti-inflammatory capacity of ferulic acid released from wheat bran by this A. niger strain (FA-WB) were evaluated.@*METHODS@#Molecular identification techniques based on PCR analysis of specific genomic sequences were conducted; antioxidant ability was examined using oxygen radical absorbance capacity (ORAC), cellular antioxidant activity (CAA) assays, and erythrocyte hemolysis assays. RAW264.7 cells were used as a model to detect anti-inflammatory activity.@*RESULTS@#The filamentous fungal isolate was identified to be A. niger. ORAC and CAA assay showed that FA-WB had better antioxidant activity than that of the ferulic acid standard. The erythrocyte hemolysis assay results suggested that FA-WB could attenuate AAPH-induced oxidative stress through inhibition of reactive oxy gen species (ROS) generation. FA-WB could significantly restore the AAPH-induced increase in intracellular antioxidant enzyme activities to normal levels as well as inhibit the intracellular malondialdehyde formation. TNF-a, IL-6, and NO levels indicated that FA-WB can inhibit the inflammation induced by lipopolysaccharide (LPS).@*CONCLUSION@#Ferulic acid released from wheat bran by a new strain of A. niger had good anti-inflammatory activity and better antioxidant ability than standard ferulic acid.
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Animales , Humanos , Ratones , Antiinflamatorios , Metabolismo , Farmacología , Antioxidantes , Metabolismo , Farmacología , Aspergillus niger , Genética , Metabolismo , Ácidos Cumáricos , Metabolismo , Farmacología , ADN de Hongos , Fibras de la Dieta , Microbiología , Eritrocitos , Metabolismo , Fermentación , Células Hep G2 , Interleucina-6 , Metabolismo , Lipopolisacáridos , Farmacología , Ovinos , Factor de Necrosis Tumoral alfa , MetabolismoRESUMEN
Acrolein, known as one of the most common reactive carbonyl species, is a toxic small molecule affecting human health in daily life. This study is focused on the scavenging abilities and mechanism of ferulic acid and some other phenolic acids against acrolein. Among the 13 phenolic compounds investigated, ferulic acid was found to have the highest efficiency in scavenging acrolein under physiological conditions. Ferulic acid remained at (3.04±1.89)% and acrolein remained at (29.51±4.44)% after being incubated with each other for 24 h. The molecular mechanism of the detoxifying process was also studied. Detoxifying products, namely 2-methoxy-4-vinylphenol (product 21) and 5-(4-hydroxy-3-methoxyphenyl)pent-4-enal (product 22), were identified though nuclear magnetic resonance (NMR) and gas chromatography-mass spectrometry (GC-MS), after the scavenging process. Ferulic acid showed significant activity in scavenging acrolein under physiological conditions. This study indicates a new method for inhibiting damage from acrolein.
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Acroleína/toxicidad , Ácidos Cumáricos/farmacología , Glutatión/fisiología , Hidroxibenzoatos/farmacología , Espectroscopía de Resonancia Magnética , Relación Estructura-ActividadRESUMEN
Background: Aspergillus ochraceus was isolated from coffee pulp and selected as an interesting hydroxycinnamoyl esterase strain producer, using an activity microplate high-throughput screening method. In this work, we purified and characterized a new type C A. ochraceus feruloyl esterase (AocFaeC), which synthesized specifically butyl hydroxycinnamates in a ternary solvent system. Results: AocFaeC was produced by solid state fermentation, reaching its maximal activity (1.1 U/g) after 48 h of culture. After purification, the monomeric protein (34 kDa) showed a specific activity of 57.9 U/mg towards methyl ferulate. AocFaeC biochemical characterization confirmed its identity as a type C feruloyl esterase and suggested the presence of a catalytic serine in the active site. Its maximum hydrolytic activity was achieved at 40°C and pH 6.5 and increased by 109 and 77% with Ca2+ and Mg2+, but decreased by 90 and 45% with Hg2+ and Cu2+, respectively. The initial butyl ferulate synthesis rate increased from 0.8 to 23.7 nmol/min after transesterification condition improvement, using an isooctane:butanol:water ternary solvent system, surprisingly the synthesis activity using other alcohols was negligible. At these conditions, the synthesis specific activities for butyl p-coumarate, sinapinate, ferulate, and caffeate were 87.3, 97.6, 168.2, and 234 U/µmol, respectively. Remarkably, AocFaeC showed 5 folds higher butyl caffeate synthesis rate compared to type B Aspergillus niger feruloyl esterase, a well-known enzyme for its elevated activity towards caffeic acid esters. Conclusions: Type C feruloyl esterase from A. ochraceus is a butanol specific biocatalyst for the synthesis of hydroxycinnamates in a ternary solvent system
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Aspergillus ochraceus/enzimología , Hidrolasas de Éster Carboxílico/metabolismo , Ácidos Cumáricos/síntesis química , Solventes , Espectrofotometría , Hidrolasas de Éster Carboxílico/aislamiento & purificación , Cromatografía , Café , Butanoles , Electroforesis , FermentaciónRESUMEN
ABSTRACT Propolis produced by selected bees Apis mellifera were collected from March to June of 2013 and in March of 2015 and analyzed in order to evaluate the influence of climate, colony of origin, and food supplementation of colonies on the content of total phenolic and flavonoid by chromatographic analysis and antioxidant activity by radical scavenging of 2,2-diphenyl-1-picrylhydrazyl hydrate (DPPH) and 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulphonic acid) (ABTS) and ferric reducing antioxidant power (FRAP) methods. The Principal Component Analysis (PCA) was carried out with propolis collected in 2013 and two clusters were formed. Propolis produced in the months of March and April showed a higher content of total phenolic compounds (TPC) and antioxidant capacity than those produced in May and June. The results of PCA obtained from samples collected in March of 2013 and 2015 showed two clusters, and propolis collected in 2015 were more bioactive and presented a higher content of TPC. The chromatographic analysis of extracts allowed the identification of phenolic acids p-coumaric, ferulic and caffeic with similar chemical profiles that could be closely related to the botanical origin of propolis. It can be concluded that the season and food supplementation of colonies influenced the chemical composition and the biological activity of samples analysed.
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Animales , Própolis/química , Estaciones del Año , Abejas/fisiología , Suplementos Dietéticos , Hidroxibenzoatos/análisis , Antioxidantes/análisis , Valores de Referencia , Temperatura , Flavonoides/análisis , Ácidos Cafeicos/análisis , Análisis Multivariante , Reproducibilidad de los Resultados , Cromatografía Líquida de Alta Presión , Ácidos Cumáricos/análisis , Análisis de Componente Principal , Indicadores y ReactivosRESUMEN
Ephedra tweediana (Ephedraceae), conocida como tramontana, es empleada en la medicina popular como antiasmático. Se analizaron comparativamente los extractos acuosos de los tallos herbáceos, lignificados y partes subterráneas, provenientes de ejemplares femeninos y masculinos. En los tallos lignificados se determinó la presencia de proapigeninidina (cuya identidad fue corroborada por comparación con un testigo de apigeninidina sometido a estudios de FAB-MS, UV, HPLC, 1H-NMR y espectroscopía IR); mientras que en los órganos subterráneos se determinó la presencia de proapigeninidina y propelargonidina (la identidad de la pelargonidina fue establecida por comparación con un testigo de pelargonidina sometido a estudios de TLC/HPTLC y espectroscopía UV-Visible). Estos compuestos no se observaron en los tallos herbáceos. Los tallos herbáceos presentaron las mayores concentraciones de flavonoides y ácidos hidroxicinámicos totales. Los órganos subterráneos presentaron la mayor concentración de taninos y proantocianidinas. En los tallos herbáceos se detectó una reacción fuertemente positiva para flavonoides. No se observó reacción positiva para proantocianidinas. En el tallo aéreo lignificado se observó una reacción positiva para flavonoides y proantocianidinas a nivel de la peridermis. En los órganos subterráneos, los flavonoides y proantocianidinas se localizaron principalmente en los tejidos más externos. Este trabajo constituye el primer aporte a la dinámica de polifenoles de E. tweediana.
Ephedra tweediana (Ephedraceae), known as "tramontana" is used in folk medicine as antiasthmatic. Aqueous extracts obtained from young stems, woody stems and underground parts were analyzed and compared. In lignified stems was detected proapigeninidin (whose identity was confirmed by comparison with a control apigeninidin subjected to FAB-MS, UV, HPLC, 1H-NMR and IR spectroscopy), while underground organs were detected proapigeninidin and propelargonidin (by comparison with a control pelargonidin whose identity was established by studies of TLC/HPTLC, and UV-visible spectroscopy). These compounds were not observed in the herbaceous stems. The herbaceous stems had the highest concentrations of total flavonoids and hydroxycinnamic acids. Uderground organs had the highest concentration of tannins and proanthocyanidins. In herbaceous stems a strong positive reaction for flavonoids was detected. No positive reaction was observed for proanthocyanidins. In the periderm of woody aerial stem a positive reaction for flavonoids and proanthocyanidins were observed. In the underground organs, flavonoids and proanthocyanidins were located mainly in the more external tissues. This work is the first contribution to the dynamic of E. tweediana polyphenols.
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Humanos , Ácidos Cumáricos/análisis , Ephedra/química , Flavonoides/análisis , Fenoles/análisis , Extractos Vegetales/química , Cromatografía de Gases/métodos , Proantocianidinas/análisisRESUMEN
Previous studies have reported on the glucose and lipid-lowering effects of ferulic acid (FA) but its anti-obesity potential has not yet been firmly established. This study investigated the possible anti-obesitogenic effects of FA in mice fed a high-fat diet (HFD) for 15 weeks. To assess the antiobesity potential of FA, 32 male Swiss mice, weighing 20–25 g (n=6–8 per group) were fed a normal diet (ND) or HFD, treated orally or not with either FA (10 mg/kg) or sibutramine (10 mg/kg) for 15 weeks and at the end of this period, the body weights of animals, visceral fat accumulation, plasma levels of glucose and insulin hormone, amylase and lipase activities, the satiety hormones ghrelin and leptin, and tumor necrosis factor-α (TNF-α) and monocyte chemoattractant protein-1 (MCH-1) were analyzed. Results revealed that FA could effectively suppress the HFD-associated increase in visceral fat accumulation, adipocyte size and body weight gain, similar to sibutramine, the positive control. FA also significantly (P<0.05) decreased the HFD-induced elevations in serum lipid profiles, amylase and lipase activities, and the levels of blood glucose and insulin hormone. The markedly elevated leptin and decreased ghrelin levels seen in HFD-fed control mice were significantly (P<0.05) reversed by FA treatment, almost reaching the values seen in ND-fed mice. Furthermore, FA demonstrated significant (P<0.05) inhibition of serum levels of inflammatory mediators TNF-α, and MCH-1. These results suggest that FA could be beneficial in lowering the risk of HFD-induced obesity via modulation of enzymatic, hormonal and inflammatory responses.
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Animales , Masculino , Ratones , Fármacos Antiobesidad/farmacología , Ácidos Cumáricos/farmacología , Ciclobutanos/farmacología , Grasa Intraabdominal/efectos de los fármacos , Obesidad/tratamiento farmacológico , Tejido Adiposo/patología , Dieta Alta en Grasa , Modelos Animales de Enfermedad , Obesidad/patologíaRESUMEN
Ferulic acid (FA) is an active component of herbal medicines. One of the best documented activities of FA is its antioxidant property. Moreover, FA exerts antiallergic, anti-inflammatory, and hepatoprotective effects. However, the metabolic pathways of FA in humans remain unclear. To identify whether human CYP or UGT enzymes are involved in the metabolism of FA, reaction phenotyping of FA was conducted using major CYP-selective chemical inhibitors together with individual CYP and UGT Supersomes. The CYP- and/or UGT-mediated metabolism kinetics were examined simultaneously or individually. Relative activity factor and total normalized rate approaches were used to assess the relative contributions of each major human CYPs towards the FA metabolism. Incubations of FA with human liver microsomes (HLM) displayed NADPH- and UDPGA-dependent metabolism with multiple CYP and UGT isoforms involved. CYPs and UGTs contributed equally to the metabolism of FA in HLM. Although CYP1A2 and CYP3A4 appeared to be the major contributors in the CYP-mediated clearance, their contributions to the overall clearance are still minor (< 25%). As a constitute of many food and herbs, FA poses low drug-drug interaction risk when co-administrated with other herbs or conventional medicines because multiple phase I and phase II enzymes are involved in its metabolism.
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Humanos , Ácidos Cumáricos , Química , Metabolismo , Sistema Enzimático del Citocromo P-450 , Química , Metabolismo , Medicamentos Herbarios Chinos , Metabolismo , Glucuronosiltransferasa , Química , Metabolismo , Cinética , Medicina Tradicional China , Microsomas Hepáticos , QuímicaRESUMEN
ABSTRACT The development and validation of a simple and efficient method for the quantification of ferulic acid in poly (D,L-lactide-co-glycolide) (PLGA) nanoparticles coated with chitosan (CS) by reverse phase high performance liquid chromatography coupled to photodiode array detection was described. For the chromatographic analysis, a reverse phase C-18 column was used, mobile phase consisting of acetonitrile and 0.5% acetic acid (37:63, v/v), isocratically eluted at a flow rate of 1 mL/min. Drug determination was performed at 320 nm. The method was validated in terms of the selectivity, linearity, precision, accuracy, robustness, limits of detection and quantification. The method was linear in the range of 10 to 100 µg/mL (r=0.999) and presented limit of detection and quantification of 102 ng/mL and 310 ng/mL, respectively. The method was precise (intra and inter-day) based on relative standard deviation values (less than 3.20%). The recovery was between 101.06 and 102.10%. Robustness was demonstrated considering change in mobile phase proportion. Specificity assay showed no interference from the components of nanoparticles or from the degradation products derived from acidic and oxidative conditions. The proposed method was suitable to be applied in determining the encapsulation efficiency of ferulic acid in PLGA-CS nanoparticles and can be employed as stability indicating one.
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Cromatografía Líquida de Alta Presión/métodos , Quitosano/farmacología , Nanopartículas/clasificación , Ácidos Cumáricos/farmacología , Estudio de Validación , Antioxidantes/clasificaciónRESUMEN
Background: Brettanomyces bruxellensis is an important spoilage yeast in the winemaking process. The capacity of this yeast to generate an undesired off-flavor constitutes a significant loss in the Chilean wine industry. Results: The proteomic profile of B. bruxellensis in the presence of p-coumaric acid was determined by 2D gel electrophoresis, gel image analysis and differential spot selection. A set of 41 proteins showed a differential accumulation of ±2 and a p-value <0.0001. The homology sequence analysis was performed using the databases available. Differential proteins belonged to the categories of 'energy production and conversion' and 'amino acid transport and metabolism'. Conclusions: The proteomic profile of B. bruxellensis cultivated in the presence of p-coumaric acid in synthetic wine, agrees with the hypothesis of metabolic flux regulation, allowing a better conditioning to an adverse environment. This study involved the translational level of B. bruxellensis in the production of ethylphenols and corroborated that this yeast presented an advantage in these stress conditions. Thus, this work will allow an understanding of the regulation and processes involved in the production of ethyl-derivate compounds by B. bruxellensis. Furthermore, it allows the development of newer and better techniques for spoilage yeast control.
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Vino , Ácidos Cumáricos/metabolismo , Proteoma , Brettanomyces/metabolismo , Fenoles , Levaduras , Electroforesis en Gel Bidimensional , Electroforesis , Industria VitivinícolaRESUMEN
Fractionation of the EtOH extract from aerial parts of Baccharis uncinella C. DC. (Asteraceae) led to isolation of caffeic and ferulic acids, which were identified from spectroscopic and spectrometric evidence. These compounds exhibit antioxidant and anti-inflammatory properties and have been shown to be effective in the prevention/treatment of metabolic syndrome. This study investigated whether the combined treatment of caffeic and ferulic acids exhibits a more significant beneficial effect in a mouse model with metabolic syndrome. The combination treatment with caffeic and ferulic acids was tested for 60 days in C57 mice kept on a high-fat (40%) diet. The data obtained indicated that treatment with caffeic and ferulic acids prevented gain in body weight induced by the high-fat diet and improved hyperglycemia, hypercholesterolemia and hypertriglyceridemia. The expression of a number of metabolically relevant genes was affected in the liver of these animals, showing that caffeic and ferulic acid treatment results in increased cholesterol uptake and reduced hepatic triglyceride synthesis in the liver, which is a likely explanation for the prevention of hepatic steatosis. In conclusion, the combined treatment of caffeic and ferulic acids displayed major positive effects towards prevention of multiple aspects of the metabolic syndrome and liver steatosis in an obese mouse model.