RESUMEN
AIMS: This study aimed to investigate the effects of Umbelliferone (UMB) on the inflammation underlying alveolar bone resorption in mouse periodontitis. METHODS: Male Swiss mice subjected to a ligature of molars were grouped as non-treated (NT), received UMB (15, 45, or 135 mg/kg) or saline daily for 7 days, respectively, and were compared with naïve mice as control. Gingival tissues were evaluated by myeloperoxidase (MPO) activity and interleukin-1ß level by ELISA. The bone resorption was directly assessed on the region between the cement-enamel junction and the alveolar bone crest. Microscopically, histomorphometry of the furcation region, immunofluorescence for nuclear factor-kappa B (NF-ĸB), and immunohistochemistry for tartrate-resistant acid phosphatase (TRAP), and cathepsin K (CTSK) were performed. Systemically, body mass variation and leukogram were analyzed. RESULTS: Periodontitis significantly increased MPO activity, interleukin-1ß level, and NF-ĸB+ immunofluorescence, and induced severe alveolar bone and furcation resorptions, besides increased TRAP+ and CTSK+ cells compared with naïve. UMB significantly prevented the inflammation by reducing MPO activity, interleukin-1ß level, and NF-ĸB+ intensity, besides reduction of resorption of alveolar bone and furcation area, and TRAP+ and CTSK+ cells compared with the NT group. Periodontitis or UMB treatment did not affect the animals systemically. CONCLUSION: UMB improved periodontitis by reducing inflammation and bone markers.
Asunto(s)
Pérdida de Hueso Alveolar , Interleucina-1beta , Periodontitis , Umbeliferonas , Animales , Masculino , Ratones , Pérdida de Hueso Alveolar/prevención & control , Pérdida de Hueso Alveolar/patología , Pérdida de Hueso Alveolar/tratamiento farmacológico , Periodontitis/tratamiento farmacológico , Periodontitis/patología , Umbeliferonas/uso terapéutico , Umbeliferonas/farmacología , Osteoclastos/efectos de los fármacos , Osteoclastos/patología , FN-kappa B/metabolismo , Fosfatasa Ácida Tartratorresistente/metabolismo , Peroxidasa , Inflamación , Catepsina K , Ligadura , Encía/patología , Encía/efectos de los fármacosRESUMEN
The aim of this work was to compare the anti-inflammatory and antioxidant effects of three natural coumarins: 1,2-benzopyrone, umbelliferone and esculetin. The antioxidant capacity of coumarins was evaluated using both chemical and biological in vitro assays. Chemical assays included DPPH and ABTSâ+ radical scavenging as well as ferric ion reducing ability power (FRAP) assay. Inhibition of mitochondrial ROS generation and lipid peroxidation in brain homogenates were used as biological in vitro assays. The experimental method of carrageenan-induced pleurisy in rats was used for the in vivo investigation of the anti-inflammatory activity. In silico molecular docking analysis was undertaken to predict the affinity of COX-2 to the coumarins. Considering the antioxidant capacity, esculetin was the most efficient one as revealed by all employed assays. Particularly, the mitochondrial ROS generation was totally abolished by the compound at low concentrations (IC50 = 0.57 µM). As for the anti-inflammatory effects, the COX-2 enzyme presented good affinities to the three coumarins, as revealed by the molecular docking analyses. However, considering the in vivo anti-inflammatory effects, 1,2-benzopyrone was the most efficient one in counteracting pleural inflammation and it potentiated the anti-inflammatory actions of dexamethasone. Umbelliferone and esculetin treatments failed to reduce the volume of pleural exudate. Overall, therefore, our results support the notion that this class of plant secondary metabolites displays promising effects in the prevention and/or treatment of inflammation and other diseases associated with oxidative stress, although the singularities regarding the type of the inflammatory process and pharmacokinetics must be taken into account.
Asunto(s)
Antioxidantes , Cumarinas , Ratas , Animales , Antioxidantes/farmacología , Antioxidantes/uso terapéutico , Cumarinas/farmacología , Cumarinas/uso terapéutico , Especies Reactivas de Oxígeno , Ciclooxigenasa 2/metabolismo , Simulación del Acoplamiento Molecular , Umbeliferonas/farmacología , Umbeliferonas/uso terapéutico , Inflamación/inducido químicamente , Inflamación/tratamiento farmacológico , Antiinflamatorios/farmacología , Antiinflamatorios/uso terapéutico , Extractos Vegetales/farmacologíaRESUMEN
This study investigated the in vitro and in vivo antiproliferative activity of esculetin against hepatocellular carcinoma, and clarified its potential molecular mechanisms. Cell viability was determined by the MTT (tetrazolium) colorimetric assay. In vivo antitumor activity of esculetin was evaluated in a hepatocellular carcinoma mouse model. Seventy-five C57BL/6J mice were implanted with Hepa1-6 cells and randomized into five groups (n=15 each) given daily intraperitoneal injections of vehicle (physiological saline), esculetin (200, 400, or 700 mg·kg-1·day-1), or 5-Fu (200 mg·kg-1·day-1) for 15 days. Esculetin significantly decreased tumor growth in mice bearing Hepa1-6 cells. Tumor weight was decreased by 20.33, 40.37, and 55.42% with increasing doses of esculetin. Esculetin significantly inhibited proliferation of HCC cells in a concentration- and time-dependent manner and with an IC50 value of 2.24 mM. It blocked the cell cycle at S phase and induced apoptosis in SMMC-7721 cells with significant elevation of caspase-3 and caspase-9 activity, but did not affect caspase-8 activity. Moreover, esculetin treatment resulted in the collapse of mitochondrial membrane potential in vitro and in vivo accompanied by increased Bax expression and decreased Bcl-2 expression at both transcriptional and translational levels. Thus, esculetin exerted in vitro and in vivo antiproliferative activity in hepatocellular carcinoma, and its mechanisms involved initiation of a mitochondrial-mediated, caspase-dependent apoptosis pathway.
Asunto(s)
Antioxidantes/uso terapéutico , Apoptosis/efectos de los fármacos , Carcinoma Hepatocelular/tratamiento farmacológico , Neoplasias Hepáticas/tratamiento farmacológico , Potencial de la Membrana Mitocondrial/efectos de los fármacos , Umbeliferonas/uso terapéutico , Animales , Proliferación Celular/efectos de los fármacos , Supervivencia Celular/efectos de los fármacos , Masculino , Ratones , Ratones Endogámicos C57BL , Neoplasias Experimentales , Distribución AleatoriaRESUMEN
BACKGROUND: Coumarins, also known as benzopyrones, are plant-derived products with several pharmacological properties, including antioxidant and anti-inflammatory activities. Based on the wide distribution of coumarin derivatives in plant-based foods and beverages in the human diet, our objective was to evaluate both the antioxidant and intestinal anti-inflammatory activities of six coumarin derivatives of plant origin (scopoletin, scoparone, fraxetin, 4-methyl-umbeliferone, esculin and daphnetin) to verify if potential intestinal anti-inflammatory activity was related to antioxidant properties. METHODS: Intestinal inflammation was induced by intracolonic instillation of TNBS in rats. The animals were treated with coumarins by oral route. The animals were killed 48 h after colitis induction. The colonic segments were obtained after laparotomy and macroscopic and biochemical parameters (determination of glutathione level and myeloperoxidase and alkaline phosphatase activities) were evaluated. The antioxidant properties of these coumarins were examined by lipid peroxidation and DPPH assays. RESULTS: Treatment with esculin, scoparone and daphnetin produced the best protective effects. All coumarin derivatives showed antioxidant activity in the DPPH assay, while daphnetin and fraxetin also showed antioxidant activity by inhibiting lipid peroxidation. Coumarins, except 4-methyl-umbeliferone, also showed antioxidant activity through the counteraction of glutathione levels or through the inhibition of myeloperoxidase activity. DISCUSSION: The intestinal anti-inflammatory activity of coumarin derivatives were related to their antioxidant properties, suggesting that consumption of coumarins and/or foods rich in coumarin derivatives, particularly daphnetin, esculin and scoparone, could prevent intestinal inflammatory disease.
Asunto(s)
Antiinflamatorios/farmacología , Antioxidantes/farmacología , Colitis/metabolismo , Colon/efectos de los fármacos , Cumarinas/farmacología , Fitoterapia , Extractos Vegetales/farmacología , Fosfatasa Alcalina/metabolismo , Animales , Antiinflamatorios/uso terapéutico , Antioxidantes/uso terapéutico , Compuestos de Bifenilo/metabolismo , Colitis/etiología , Colitis/prevención & control , Colon/metabolismo , Cumarinas/uso terapéutico , Esculina/farmacología , Esculina/uso terapéutico , Glutatión/metabolismo , Inflamación/etiología , Inflamación/metabolismo , Inflamación/prevención & control , Enfermedades Inflamatorias del Intestino/metabolismo , Enfermedades Inflamatorias del Intestino/prevención & control , Peroxidación de Lípido/efectos de los fármacos , Masculino , Estrés Oxidativo/efectos de los fármacos , Peroxidasa/metabolismo , Picratos/metabolismo , Extractos Vegetales/uso terapéutico , Ratas , Ratas Wistar , Umbeliferonas/farmacología , Umbeliferonas/uso terapéuticoRESUMEN
The aim of the present study was to compare the effects of the 4-methylesculetin with those produced by prednisolone and sulphasalazine and to elucidate the mechanisms involved in its action. Colitis was induced in rat by instillation of trinitrobenzenesulphonic acid (TNBS). The colon damage was evaluated using macroscopic, microscopic and biochemical analysis. In addition, in vitro studies were performed to evaluate cytokine production in cell cultures using the murine macrophage cell line RAW264.7, mouse splenocytes and the human colonic epithelial cell line Caco-2. 4-Methylesculetin produced a reduction of the macroscopic damage score and the recovery of the intestinal cytoarchitecture. These effects were associated with a prevention of the GSH depletion and an inhibition in AP activity. After colitis relapse, 4-methylesculetin improved the colonic inflammatory status as evidenced by histological findings, with a reduction in apoptosis, as well as biochemically by inhibition of colonic myeloperoxidase, alkaline phosphatase and metalloproteinase 9 activities. Paired with this inhibitive activity, there was a decrease in malondialdehyde content and in IL-1ß levels. In vitro assays revealed that 4-methylesculetin promoted an inhibition in IL-1ß, IL-8, IL-2 and IFN-γ production in cell cultures. In conclusion, 4-methylesculetin showed similar efficacy to that obtained with either prednisolone or sulphasalazine, both in the acute phase of colitis as well as following a curative protocol. The intestinal anti-inflammatory activity by 4-methylesculetin is likely related to its ability in reduce colonic oxidative stress and inhibit pro-inflammatory cytokine production.
Asunto(s)
Antiinflamatorios/farmacología , Apoptosis/efectos de los fármacos , Colitis/patología , Cumarinas/farmacología , Prednisolona/farmacología , Sulfasalazina/farmacología , Umbeliferonas/farmacología , Fosfatasa Alcalina/antagonistas & inhibidores , Fosfatasa Alcalina/metabolismo , Animales , Antiinflamatorios/química , Línea Celular , Colitis/inducido químicamente , Cumarinas/química , Citocinas/metabolismo , Regulación de la Expresión Génica/efectos de los fármacos , Glutatión/metabolismo , Humanos , Masculino , Malondialdehído/metabolismo , Metaloproteinasa 9 de la Matriz/metabolismo , Ratones , Peroxidasa/metabolismo , Ratas , Ratas Wistar , Recurrencia , Ácido Trinitrobencenosulfónico/toxicidad , Umbeliferonas/química , Umbeliferonas/uso terapéuticoRESUMEN
7-Hydroxycoumarin (umbelliferone, 1), the main metabolite of coumarin, has been reported to produce potent antinociceptive effects in animal models of pain. However, the biochemical events involved in antinociception mediated by 1 are currently not well understood. In the present study, the mechanisms by which 1 exerts its pharmacological actions were investigated. Acute pretreatment of mice with 1 produced a long-lasting antinociceptive effect against complete Freund's adjuvant (CFA)-induced hyperalgesia. The subchronic administration of 1 inhibited CFA-induced hyperalgesia and paw edema, while it did not cause any apparent toxicity. Another set of experiments showed that 1 inhibited carrageenan-induced mechanical hyperalgesia, but not mechanical hyperalgesia induced by prostaglandin E(2) (PGE(2)), suggesting that it acts upstream of PGE(2.) Treatment with 1 was able to prevent the plantar tissue neutrophil influx induced by local inflammatory stimuli. In addition, 1 exhibited inhibitory effects on the release of hyperalgesic cytokines (TNF-α and IL-1ß) and the production of PGE(2), a directly acting hyperalgesic mediator. The present results suggest that the antinociceptive effect of 1 is correlated with the inhibition of neutrophil migration, cytokine release, and PGE(2) production and are supportive of the further investigation of the therapeutic potential of 1 to control inflammatory pain.
Asunto(s)
Analgésicos/farmacología , Dolor/tratamiento farmacológico , Umbeliferonas/farmacología , Analgésicos/química , Analgésicos/metabolismo , Analgésicos/uso terapéutico , Animales , Brasil , Dinoprostona/farmacología , Edema/inducido químicamente , Edema/tratamiento farmacológico , Adyuvante de Freund/farmacología , Ratones , Modelos Animales , Estructura Molecular , Neutrófilos/efectos de los fármacos , Neutrófilos/fisiología , Dolor/inducido químicamente , Dimensión del Dolor , Factor de Necrosis Tumoral alfa/farmacología , Umbeliferonas/química , Umbeliferonas/uso terapéuticoRESUMEN
OBJECTIVES: In the present study we investigated the antinociceptive, anti-inflammatory and antipyretic effects of 7-hydroxycoumarin (7-HC) in animal models. METHODS: The effects of oral 7-HC were tested against acetic acid-induced writhing, formalin test, tail flick test, complete Freund's adjuvant (CFA)-induced hypernociception, carrageenan-induced paw oedema, lipopolysaccharide-induced fever and the rota rod test. KEY FINDINGS: 7-HC (3-60 mg/kg) produced a dose-related antinociception against acetic acid-induced writhing in mice and in the formalin test. In contrast, treatment with 7-HC did not prevent thermal nociception in the tail flick test. A single treatment with 7-HC, 60 mg/kg, produced a long-lasting antinociceptive effect against CFA-induced hypernociception, a chronic inflammatory pain stimulus. Notably, at 60 mg/kg per day over 4 days the administration of 7-HC produced a continuous antinociceptive effect against CFA-induced hypernociception. 7-HC (30-120 mg/kg) produced anti-inflammatory and antipyretic effects against carrageenan-induced inflammation and lipopolysaccharide-induced fever, respectively. Moreover, 7-HC was found to be safe with respect to ulcer induction. In the rota rod test, 7-HC-treated mice did not show any motor performance alterations. CONCLUSIONS: The prolonged antinociceptive and anti-inflammatory effects of 7-HC, in association with its low ulcerogenic activity, indicate that this molecule might be a good candidate for development of new drugs for the control of chronic inflammatory pain and fever.
Asunto(s)
Antiinflamatorios no Esteroideos/uso terapéutico , Inflamación/tratamiento farmacológico , Dolor/tratamiento farmacológico , Umbeliferonas/uso terapéutico , Enfermedad Aguda , Administración Oral , Animales , Antiinflamatorios no Esteroideos/administración & dosificación , Antiinflamatorios no Esteroideos/efectos adversos , Antiinflamatorios no Esteroideos/farmacología , Enfermedad Crónica , Modelos Animales de Enfermedad , Relación Dosis-Respuesta a Droga , Edema/tratamiento farmacológico , Fiebre/tratamiento farmacológico , Masculino , Ratones , Actividad Motora/efectos de los fármacos , Dimensión del Dolor , Ratas , Ratas Wistar , Umbeliferonas/administración & dosificación , Umbeliferonas/efectos adversos , Umbeliferonas/farmacologíaRESUMEN
The therapeutic effects of umbelliferone (30, 60 and 90 mg/kg), a coumarin isolated from Typha domingensis (Typhaceae) were investigated in a mouse model of bronchial asthma. BALB/c mice were immunized and challenged by nasal administration of ovalbumin. Treatment with umbelliferone (60 and 90 mg/kg) caused a marked reduction of cellularity and eosinophil numbers in bronchoalveolar lavage fluids from asthmatic mice. In addition, a decrease in mucus production and lung inflammation were observed in mice treated with umbelliferone. A reduction of IL-4, IL-5, and IL-13, but not of IFN-gamma, was found in bronchoalveolar lavage fluids of mice treated with umbelliferone, similar to that observed with dexamethasone. The levels of ovalbumin-specific IgE were not significantly altered after treatment with umbelliferone. In conclusion, our results demonstrate that umbelliferone attenuates the alteration characteristics of allergic airway inflammation. The investigation of the mechanisms of action of this molecule may contribute for the development of new drugs for the treatment of asthma.
Asunto(s)
Antiasmáticos/farmacología , Asma/inmunología , Neumonía/inmunología , Umbeliferonas/farmacología , Animales , Antiasmáticos/química , Antiasmáticos/uso terapéutico , Asma/patología , Líquido del Lavado Bronquioalveolar/citología , Líquido del Lavado Bronquioalveolar/inmunología , Modelos Animales de Enfermedad , Relación Dosis-Respuesta a Droga , Interleucina-13/inmunología , Interleucina-4/inmunología , Interleucina-5/inmunología , Masculino , Ratones , Ratones Endogámicos BALB C , Estructura Molecular , Ovalbúmina/inmunología , Neumonía/patología , Umbeliferonas/química , Umbeliferonas/uso terapéuticoRESUMEN
Introducción: Las cumarinas son compuestos ampliamente distribuidos en la naturaleza. En el humano el principal metabolito de la cumarina es la 7-hidroxicumarina. Estos compuestos presentan actividad antitumoral in vitro e in vivo. Previamente reportamos que la cumarina y la 7-hidroxicumarina ejercen actividad citostática en líneas de adenocarcinoma; sin embargo, se desconocen los aspectos relacionados a la fase del ciclo celular en la que estos compuestos pueden actuar.Objetivo: Determinar el efecto de la cumarina y la 7-hidroxicumarina en la transición del ciclo celular en líneas de adenocarcinoma pulmonar.Material y métodos: Tres líneas de adenocarcinoma pulmonar (SK-LU-1, 1.3.15 y 3A5A) y células mononucleares de individuos sanos estimuladas con fitohemaglutinina, se incubaron por 24h y 40h respectivamente, con 100 µg/mL de cumarina o 7-hidroxicumarina. Las perturbaciones en el ciclo celular se evaluaron por medio de la determinación de contenido de DNA por citometría de flujo.Resultados: Posterior al tratamiento, las líneas celulares presentaron acumulación significativa de células en la fase G0/G1, disminuyendo proporcionalmente la población de células en la fase S. En cambio, las células mononucleares no presentaron cambios significativos después del tratamiento.Conclusiones: Estos resultados demuestran que los compuestos cumarínicos causan un arresto de las células tumorales en la fase G0/G1. El conocimiento de las perturbaciones en el ciclo celular, inducidas por estos productos, puede facilitar la elaboración de esquemas de tratamiento mediante combinaciones con otros fármacos que arresten en distinta(s) fase(s) del ciclo. Además, esta metodología permite evaluar la susceptibilidad de las células tumorales a los agentes quimioterapéuticos.