RESUMEN
Quinoa is one of the crops well-adapted to high altitude regions that can grow relatively well under drought, humid, and high UV radiation conditions. This study was performed to investigate the effects of gamma-radiation on quinoa. Seeds were treated with various doses of 50 Gy, 100 Gy, 200 Gy, 300 Gy, 400 Gy, 600 Gy, 800 Gy, and 1000 Gy. We investigated germination, as well as plant height, chlorophyll content, and normalized difference vegetation index (NDVI) at 0, 30, 44, 58, and 88 days after transplanting (DAT) and panicle weight at 88 DAT. The plants grown from the seeds treated at radiation doses greater than 200 Gy showed reduced values in most growth and physiological characteristics. The germination rate and germination speed were higher in the 50 Gy-treated seeds than in 0 Gy-treated (control) seeds. Plant height and panicle weight were highest in the plants from 50 Gy-treated seeds. Chlorophyll content was higher in all treated samples than in the controls. NDVI value showed the highest value in 0 Gy controls and plants treated with 50 Gy. The antioxidant activity was also higher in the plants from the seeds treated with 50 Gy and 100 Gy, showing a steady increase as the radiation dose increased even at 200 Gy. The plants from seeds treated with 0 Gy showed higher expression of proteins related to photorespiration and tubulin chains. The plants from seeds treated with 50 Gy induced more stress-responsive proteins.
Asunto(s)
Chenopodium quinoa , Chenopodium quinoa/metabolismo , Clorofila/metabolismo , Rayos gamma , Semillas/metabolismo , Semillas/efectos de la radiaciónRESUMEN
Global warming has an impact on crop growth and development. Flowering time is particularly sensitive to environmental factors such as day length and temperature. In this study, we investigated the effects of global warming on flowering using an open-top Climatron chamber, which has a higher temperature and CO2 concentration than in the field. Two different soybean cultivars, Williams 82 and IT153414, which exhibited different flowering times, were promoted flowering in the open-top Climatron chamber than in the field. We more specifically examined the expression patterns of soybean flowering genes on the molecular level under high-temperature conditions. The elevated temperature induced the expression of soybean floral activators, GmFT2a and GmFT5a as well as a set of GmCOL genes. In contrast, it suppressed floral repressors, E1 and E2 homologs. Moreover, high-temperature conditions affected the expression of these flowering genes in a day length-independent manner. Taken together, our data suggest that soybean plants properly respond and adapt to changing environments by modulating the expression of a set of flowering genes in the photoperiod pathway for the successful production of seeds and offspring.
Asunto(s)
Perfilación de la Expresión Génica/métodos , Glycine max/crecimiento & desarrollo , Proteínas de Plantas/genética , Flores/genética , Flores/crecimiento & desarrollo , Regulación de la Expresión Génica de las Plantas , Calentamiento Global , Calor , Fotoperiodo , Reacción en Cadena en Tiempo Real de la Polimerasa , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa , Glycine max/genética , Estrés FisiológicoRESUMEN
Phospholipids (PLs) in 57 varieties of soybeans were profiled by high-performance liquid chromatography-electrospray ionization-tandem mass spectrometry and principal component analysis (PCA) to discriminate PL-rich soybeans. The PL calibration curves showed linearity with correlation coefficients >0.9964. The recoveries at 5 mg/L spiked level ranged from 72.8 to 86.7% and those at 12.5 mg/L from 78.2 to 85.1%. The repeatability at a 5 mg/L spiked level ranged from 2.5 to 7.0% and those at 12.5 mg/L from 1.2 to 3.9%. The average total PL content in the 57 soybean varieties was about 35.3 mg/kg. The total PL content was the highest in Aodaiz (35, 48.7 ± 1.4 mg/kg) and the lowest in Poongsannamul (56, 16.0 ± 0.7 mg/kg). The PCA showed that RS-78sun (42), Gyeongsang #1 (3) and Aodaiz (35) are the most improved varieties of the investigated 57 varieties from the viewpoint of PL content.
Asunto(s)
Cromatografía Líquida de Alta Presión/métodos , Glycine max/química , Fosfolípidos/análisis , Espectrometría de Masas en Tándem/métodos , Modelos Lineales , Fosfolípidos/química , Análisis de Componente Principal , Reproducibilidad de los Resultados , República de Corea , Semillas/químicaRESUMEN
Anthocyanins are known to have antioxidant and antiinflammatory effects. We hypothesized that anthocyanins would enhance wound healing in Sprague-Dawley rats. The purpose of this study was to evaluate our hypothesis and investigate the mechanism of wound healing enhancement. The cytoprotective effect of an immortalized epidermal keratinocyte cell line (HaCaT) and human neonatal dermal fibroblasts in response to various concentrations of anthocyanins was determined. Vascular endothelial growth factor (VEGF) and thrombospondin 1 (TSP1) of HaCaT were measured by Western blot analysis. Anthocyanins were applied to the wounds in rats, and the healing ratio was calculated. Tissue VEGF, TSP1, CD31, nuclear factor-κB, and phosphorylation of IκBα were measured. The viability of the HaCaT cell line and human neonatal dermal fibroblasts increased under cytotoxicity by H2O2 in the anthocyanin-treated groups. The VEGF in the anthocyanin-treated groups increased, whereas TSP1 decreased. Wounds in the experimental groups healed faster, and VEGF and CD31 increased in the experimental groups, whereas TSP1 decreased. Anthocyanins inhibited the translocation of nuclear factor-κB (p65) from cytosol to nucleus and also prevented the phosphorylation of IκBα. Anthocyanins enhance wound healing through a cytoprotective effect, enhancement of angiogenesis, and an antiinflammatory effect.
Asunto(s)
Antocianinas/farmacología , Antioxidantes/farmacología , Fibroblastos/efectos de los fármacos , Glycine max , Queratinocitos/efectos de los fármacos , Semillas , Cicatrización de Heridas/efectos de los fármacos , Animales , Antiinflamatorios/farmacología , Factores de Transcripción Básicos con Cremalleras de Leucinas y Motivos Hélice-Asa-Hélice/metabolismo , Biomarcadores/metabolismo , Western Blotting , Línea Celular , Citocinas/metabolismo , Citotoxinas/efectos adversos , Fibroblastos/metabolismo , Humanos , Peróxido de Hidrógeno/efectos adversos , Queratinocitos/metabolismo , Ratas , Ratas Sprague-Dawley , Factor A de Crecimiento Endotelial Vascular/metabolismo , Cicatrización de Heridas/fisiologíaRESUMEN
BACKGROUND: Soybean sprouts (Kongnamool) are one of the most popular and nutritive traditional vegetables in East Asia. Anthracnose caused by Colletotrichum gloeosporioides is one of the most serious diseases of soybean sprouts. In order to obtain basic information for breeding and/or selecting soybean genotypes with increased natural defense against anthracnose, phenolic compounds were profiled for healthy and infected soybean (Glycine max Merr.) sprouts by using high-performance liquid chromatography coupled with tandem mass spectrometry. RESULTS: Tryptophan and eight phenolic compounds (daidzin, genistin, malonyldaidzin, malonylgenistin, daidzein, glycitein, genistein and coumestrol) were determined from healthy and inoculated sprouts. Total identified phenolic content was 40.02 ± 0.03 mg kg⻹, 99.4% of which was isoflavones. CONCLUSION: The monitoring suggested that de novo induced glycitein appeared to act as a phytoalexin in the defence mechanism of the soybean sprouts against C. gloeosporioides, and constitutively formed seven phenolic components that functioned as phytoanticipins in the diseased soybean sprouts.
Asunto(s)
Colletotrichum/crecimiento & desarrollo , Glycine max/metabolismo , Glycine max/microbiología , Isoflavonas/biosíntesis , Enfermedades de las Plantas/microbiología , Plantones/metabolismo , Plantones/microbiología , Antifúngicos/análisis , Antifúngicos/química , Antifúngicos/metabolismo , Antifúngicos/farmacología , Cromatografía Líquida de Alta Presión , Colletotrichum/efectos de los fármacos , Colletotrichum/inmunología , Germinación , Glucósidos/análisis , Glucósidos/biosíntesis , Glucósidos/química , Glucósidos/farmacología , Isoflavonas/análisis , Isoflavonas/química , Isoflavonas/farmacología , Pruebas de Sensibilidad Microbiana , Micelio/efectos de los fármacos , Micelio/crecimiento & desarrollo , Micelio/inmunología , Fenoles/análisis , Fenoles/química , Fenoles/metabolismo , Fenoles/farmacología , Enfermedades de las Plantas/inmunología , Inmunidad de la Planta , República de Corea , Plantones/crecimiento & desarrollo , Plantones/inmunología , Sesquiterpenos/análisis , Sesquiterpenos/química , Sesquiterpenos/metabolismo , Sesquiterpenos/farmacología , Glycine max/crecimiento & desarrollo , Glycine max/inmunología , Espectrometría de Masa por Ionización de Electrospray , Esporas Fúngicas/efectos de los fármacos , Esporas Fúngicas/crecimiento & desarrollo , Esporas Fúngicas/inmunología , Espectrometría de Masas en Tándem , FitoalexinasRESUMEN
Ischemia-reperfusion injury is a phenomenon that occurs when tissues are subjected to ischemia for a variable period of time, and then reperfused. Inflammatory reaction has been implicated as one of the most important mechanism of ischemia-reperfusion injury. The purpose of this study was to evaluate the anti-inflammatory effects of anthocyanins from black soybean seed coat on keratinocytes in vitro and ischemia-reperfusion injury in vivo. We investigated the inhibition, by anthocyanins, of the expression of various inflammatory genes associated with ischemia-reperfusion injury in the tumor necrosis factor-alpha-treated (TNF-α) immortalized epidermal keratinocyte cell line (HaCaT). We also investigated the effects of anthocyanins on the survival of skin flaps after ischemia-reperfusion injury in the rats. According to Western blot analysis and a luciferase activity assay, anthocyanins inhibited TNF-α-induced intercellular adhesion molecule-1 and cyclooxygenase-2 (COX-2) levels through the NF-κB-dependent pathway. Administration of anthocyanins (50 and 100 mg/kg) significantly improved the flap area survival in the 10-hour ischemic model from 62% to 74.5% and 83%, respectively (P = 0.001). The related cytokines in skin flap also changed as the same pattern as in vitro. Our results indicate that anthocyanins from black soybean seed coat had anti-inflammatory effects on the HaCaT cell line and increase the survival of skin flaps through anti-inflammatory properties against ischemia-reperfusion injury.
Asunto(s)
Antocianinas/farmacología , Glycine max , Queratinocitos/efectos de los fármacos , Fitoterapia , Extractos Vegetales/farmacología , Daño por Reperfusión/prevención & control , Colgajos Quirúrgicos/irrigación sanguínea , Animales , Antocianinas/uso terapéutico , Antiinflamatorios/farmacología , Antiinflamatorios/uso terapéutico , Biomarcadores/metabolismo , Western Blotting , Línea Celular , Citocinas/metabolismo , Supervivencia de Injerto , Queratinocitos/metabolismo , Masculino , Extractos Vegetales/uso terapéutico , Distribución Aleatoria , Ratas , Ratas Sprague-Dawley , SemillasRESUMEN
The proper timing of flowering in response to environmental changes is critical for ensuring crop yields. FLOWERING LOCUS T (FT) homologs of the phosphatidylethanolamine-binding protein family play important roles as floral integrators in many crops. In soybean, we identified 17 genes of this family, and characterized biological functions in flowering for ten FT homologs. Overexpression of GmFT homologs in Arabidopsis revealed that a set of GmFT homologs, including GmFT2a/2b, GmFT3a/3b, and GmFT5a/5b, promoted flowering similar to FT; in contrast, GmFT1a/1b, GmFT4, and GmFT6 delayed flowering. Consistently, expressions of GmFT2a, GmFT2b, and GmFT5a were induced in soybean leaves in response to floral inductive short days, whereas expressions of GmFT1a and GmFT4 were induced in response to long days. Exon swapping analysis between floral activator GmFT2a and floral repressor GmFT4 revealed that the segment B region in the fourth exon is critical for their antagonistic functions. Finally, expression analysis of GmFT2a, GmFT5a, and GmFT4 in soybean accessions exhibiting various flowering times indicated that the mRNA levels of GmFT2a and GmFT5a were higher in early flowering accessions than in late-flowering accessions, while GmFT4 showed the opposite pattern. Moreover, the relative mRNA levels between GmFT2a/GmFT5a and GmFT4 was important in determining day length-dependent flowering in soybean accessions. Taken together, our results suggest that the functions of GmFT homologs have diversified into floral activators and floral repressors during soybean evolution, and the timing of flowering in response to changing day length is determined by modulating the activities of antagonistic GmFT homologs.
RESUMEN
Microglia plays a critical role in the brain and protects neuronal cells from toxins. However, over-activation of microglia leads to deleterious effects. Lipopolysaccharide (LPS) has been reported to affect neuronal cells via activation of microglia as well as directly to initiate neuroinflammation. In the present study, we evaluated the anti-inflammatory and anti-oxidative effect of anthocyanins against LPS-induced neurotoxicity in an animal model and in cell cultures. Intraperitoneal injections of LPS (250 µg/kg/day for 1 week) induce ROS production and promote neuroinflammation and neurodegeneration which ultimately leads to memory impairment. However, anthocyanins treatment at a dose of 24 mg/kg/day for 2 weeks (1 week before and 1 week co-treated with LPS) prevented ROS production, inhibited neuroinflammation and neurodegeneration, and improved memory functions in LPS-treated mice. Both histological and immunoblot analysis indicated that anthocyanins reversed the activation of JNK, prevented neuroinflammation by lowering the levels of inflammatory markers (p-NF-kB, TNF-α, and IL-1ß), and reduced neuronal apoptosis by reducing the expression of Bax, cytochrome c, cleaved caspase-3, and cleaved PARP-1, while increasing the level of survival proteins p-Akt, p-GSK3ß, and anti-apoptotic Bcl-2 protein. Anthocyanins treatment increased the levels of memory-related pre- and post-synaptic proteins and improved the hippocampus-dependent memory in the LPS-treated mice. Overall, this data suggested that consumption of naturally derived anti-oxidant agent such as anthocyanins ameliorated several pathological events in the LPS-treated animal model and we believe that anthocyanins would be a safe therapeutic agent for slowing the inflammation-induced neurodegeneration in the brain against several diseases such as Alzheimer's disease and Parkinson's disease.
Asunto(s)
Envejecimiento/patología , Antocianinas/uso terapéutico , Glucógeno Sintasa Quinasa 3 beta/metabolismo , Hipocampo/patología , Proteínas Quinasas JNK Activadas por Mitógenos/metabolismo , Memoria , Degeneración Nerviosa/tratamiento farmacológico , Proteínas Proto-Oncogénicas c-akt/metabolismo , Animales , Antocianinas/farmacología , Apoptosis/efectos de los fármacos , Línea Celular , Supervivencia Celular/efectos de los fármacos , Hipocampo/efectos de los fármacos , Hipocampo/fisiopatología , Inflamación/patología , Lipopolisacáridos , Masculino , Aprendizaje por Laberinto/efectos de los fármacos , Memoria/efectos de los fármacos , Ratones Endogámicos C57BL , Microglía/efectos de los fármacos , Microglía/metabolismo , Microglía/patología , Modelos Biológicos , Degeneración Nerviosa/patología , Degeneración Nerviosa/fisiopatología , Degeneración Nerviosa/prevención & control , Estrés Oxidativo/efectos de los fármacos , Fosforilación/efectos de los fármacos , Especies Reactivas de Oxígeno/metabolismo , Sinapsis/efectos de los fármacos , Sinapsis/metabolismo , Sinapsis/patologíaRESUMEN
Aging is a major factor involved in neurological impairments, decreased anti-oxidant activities, and enhanced neuroinflammation. D-galactose (D-gal) has been considered an artificial aging model which induces oxidative stress and inflammatory response resulting in memory and synaptic dysfunction. Dietary supplementation exerts valuable effects against oxidative stress and neuroinflammation. Polyphenolic flavonoids, such as anthocyanins, have been reported as an anti-inflammatory and anti-oxidant agents against various neurodegenerative diseases. Recently, our group reported anthocyanin neuroprotection of the developing rat brain against ethanol-induced oxidative stress and neurodegenaration and ethanol-induced neuronal apoptosis via GABAB1 receptor intracellular signaling in prenatal rat hippocampus. Here, we examined the protective effect of anthocyanin neuroprotection against D-gal-induced oxidative and inflammatory response in the hippocampus and cortex regions and explore the potential mechanism of its action. Our results indicated that anthocyanins treatment significantly improved behavioral performance of D-gal-treated rats in Morris water maze and Y-maze tests. One of the potential mechanisms of this action was decreased expression of the receptor for advance glycation end product, reduced level of reactive oxygen species (ROS) and lipid peroxidation as well as markers of the Alzheimer's disease. Furthermore, the results also indicated that anthocyanins inhibited activated astrocytes and neuroinflammation via suppression of various inflammatory markers including p-NF- K B, inducible nitric oxide synthase (iNOS), and tumor necrosis factor-alpha (TNF-α) in the hippocampus and cortex regions of D-gal-treated rats brain. Moreover, anthocyanins abrogated neuroapoptosis via C-jun N-terminal kinase (p-JNK) suppression and improved deregulated synaptic proteins including synaptophysin, synaptosomal-associated protein (SNAP)-23, SNAP-25, and phosphorylated CREB. This data suggests that anthocyanins could be a safe and promising anti-oxidant and anti-neuroinflammatory agent for age-related neurodegenerative diseases such as Alzheimer's disease.
Asunto(s)
Antocianinas/farmacología , Disfunción Cognitiva/inducido químicamente , Disfunción Cognitiva/metabolismo , Galactosa/toxicidad , Mediadores de Inflamación/metabolismo , Estrés Oxidativo/fisiología , Animales , Antocianinas/uso terapéutico , Disfunción Cognitiva/tratamiento farmacológico , Mediadores de Inflamación/antagonistas & inhibidores , Masculino , Aprendizaje por Laberinto/efectos de los fármacos , Aprendizaje por Laberinto/fisiología , Estrés Oxidativo/efectos de los fármacos , Distribución Aleatoria , Ratas , Ratas Sprague-DawleyRESUMEN
We examined the inhibition of the expression of some inflammatory genes associated with ischemia-reperfusion (I/R) injury by anthocyanins isolated from black soybean seed coat in tumor necrosis factor-alpha (TNF-alpha)-treated bovine aortic endothelial cells. In addition, its potential use on I/R-injury was investigated using rats subjected to 30-min occlusion of left descending coronary artery followed by 24-h reperfusion. Western blot analysis and luciferase activity assay showed that anthocyanins inhibited TNF-alpha-induced vascular cell adhesion molecule-1, intracellular adhesion molecule-1, and cyclooxygenase-2 levels, which is through NF-kappaB-dependent pathway. Further, anthocyanins protected myocardiac injury from I/R in rats. It is suggested that anthocyanins from black soybean seed coat can be used as a useful drug to modulate cardiovascular disorder.
Asunto(s)
Antocianinas/farmacología , Regulación de la Expresión Génica/efectos de los fármacos , Daño por Reperfusión Miocárdica/prevención & control , Daño por Reperfusión/genética , Factor de Necrosis Tumoral alfa/fisiología , Animales , Bovinos , Ciclooxigenasa 2/genética , Modelos Animales de Enfermedad , Células Endoteliales/citología , Endotelio Vascular/citología , Endotelio Vascular/efectos de los fármacos , Endotelio Vascular/metabolismo , Molécula 1 de Adhesión Intercelular/genética , Daño por Reperfusión Miocárdica/genética , Miocardio/metabolismo , Miocardio/patología , FN-kappa B/metabolismo , Ratas , Semillas , Glycine max , Molécula 1 de Adhesión Celular Vascular/genéticaRESUMEN
Isoflavones are synthesized by isoflavone synthases via the phenylpropanoid pathway in legumes. We have cloned two isoflavone synthase genes, IFS1 and IFS2, from a total of 18 soybean cultivars. The amino acid residues of the proteins that differed between cultivars were dispersed over the entire coding region. However, amino acid sequence variation did not occur in conserved domains such as the ERR triad region, except that one conserved amino acid was changed in the IFS2 protein of the GS12 cultivar (R374G) and the IFS1 proteins of the 99M06 and Soja99s65 cultivars (A109T, F105I). In three cultivars (99M06, 99M116, and Simheukpi), most of amino acid changes were such that the difference between the amino acid sequences of IFS1 and IFS2 was reduced. The expression profiles of three enzymes that convert naringenin to the isoflavone, genistein, chalcone isomerase (CHI), isoflavone synthase (IFS) and flavanone 3-hydroxylase (F3H) were examined. In general, IFS mRNA was more abundant in etiolated seedlings than mature plants whereas the levels of CHI and F3H mRNAs were similar in the two stages. During seed development, IFS was expressed a little later than CHI and F3H but expression of these three genes was barely detectable, if at all, during later seed hardening. In addition, we found that the levels of CHI, F3H, and IFS mRNAs were under circadian control. We also showed that IFS was induced by wounding and by application of methyl jasmonate to etiolated soybean seedlings.
Asunto(s)
Glycine max/enzimología , Glycine max/genética , Oxigenasas/biosíntesis , Oxigenasas/genética , Secuencia de Bases , Ritmo Circadiano , Inducción Enzimática , Regulación Enzimológica de la Expresión Génica , Genes de Plantas , Liasas Intramoleculares/biosíntesis , Oxigenasas de Función Mixta/biosíntesis , Datos de Secuencia Molecular , Polimorfismo GenéticoRESUMEN
Male germ cell apoptosis has been extensively explored in rodents. In contrast, very little is known about the susceptibility of developing germ cells to apoptosis in response to busulfan treatment. Spontaneous apoptosis of germ cells is rarely observed in the adult mouse testis, but under the experimental conditions described here, busulfan-treated mice exhibited a marked increase in apoptosis and a decrease in testis weight. TdT-mediated dUTP-X nicked end labeling analysis indicates that at one week following busulfan treatment, apoptosis was confined mainly to spermatogonia, with lesser effects on spermatocytes. The percentage of apoptosis-positive tubules and the apoptotic cell index increased in a time-dependent manner. An immediate effect was observed in spermatogonia within one week of treatment, and in the following week, secondary effects were observed in spermatocytes. RT-PCR analysis showed that expression of the spermatogonia-specific markers c-kit and Stra 8 was reduced but that Gli I gene expression remained constant, which is indicative of primary apoptosis of differentiating type A spermatogonia. Three and four weeks after busulfan treatment, RAD51 and FasL expression decreased to nearly undetectable levels, indicating that meiotic spermatocytes and post-meiotic cells, respectively, were lost. The period of germ cell depletion did not coincide with increased p53 or Fas/FasL expression in the busulfan-treated testis, although p110Rb phosphorylation and PCNA expression were inhibited. These data suggest that increased depletion of male germ cells in the busulfan-treated mouse is mediated by loss of c-kit/SCF signaling but not by p53- or Fas/FasL-dependent mechanisms. Spermatogonial stem cells may be protected from cell death by modulating cell cycle signaling such that E2F-dependent protein expression, which is critical for G1 phase progression, is inhibited.
Asunto(s)
Alquilantes/farmacología , Apoptosis/fisiología , Busulfano/farmacología , Células Germinativas , Glicoproteínas de Membrana/metabolismo , Proteínas Proto-Oncogénicas c-kit/metabolismo , Proteína p53 Supresora de Tumor/metabolismo , Receptor fas/metabolismo , Proteínas Adaptadoras Transductoras de Señales , Animales , Biomarcadores , Proteínas de Unión al ADN/metabolismo , Proteína Ligando Fas , Regulación del Desarrollo de la Expresión Génica , Células Germinativas/citología , Células Germinativas/efectos de los fármacos , Células Germinativas/fisiología , Humanos , Etiquetado Corte-Fin in Situ , Masculino , Glicoproteínas de Membrana/genética , Ratones , Tamaño de los Órganos , Antígeno Nuclear de Célula en Proliferación/metabolismo , Proteínas/metabolismo , Proteínas Proto-Oncogénicas c-kit/genética , Recombinasa Rad51 , Proteína de Retinoblastoma/metabolismo , Transducción de Señal/fisiología , Testículo/citología , Testículo/efectos de los fármacos , Testículo/metabolismo , Proteína p53 Supresora de Tumor/genética , Receptor fas/genéticaRESUMEN
Hyperglycemia, abnormal lipid and antioxidant profiles are the most usual complications in diabetes mellitus. Thus, in this study, we investigated the anti-diabetic and anti-oxidative effects of anthocyanins (ANT) from black soybean seed coats in streptozotocin (STZ)-induced diabetic rats. The administration of ANT markedly decreased glucose levels and improved heart hemodynamic function (left ventricular end diastolic pressure, +/-dp/dt parameters). ANT not only enhanced STZ-mediated insulin level decreases, but also decreased the triglyceride levels induced by STZ injection in serum. Diabetic rats exhibited a lower expression of glucose transporter 4 proteins in the membrane fractions of heart and skeletal muscle tissues, which was enhanced by ANT. In addition, ANT activated insulin receptor phosphorylation, suggesting an increased utilization of glucose by tissues. Moreover, ANT protected pancreatic tissue from STZ-induced apoptosis through regulation of caspase-3, Bax, and Bcl-2 proteins. Furthermore, ANT significantly suppressed malondialdehyde levels and restored superoxide dismutase and catalase activities in diabetic rats. Interestingly, the observed effects of ANT were superior to those of glibenclamide. Taken together, ANT from black soybean seed coat have anti-diabetic effects that are due, in part, to the regulation of glucose transporter 4 and prevention of insulin resistance and pancreatic apoptosis, suggesting a possible use as a drug to regulate diabetes.
Asunto(s)
Antocianinas/farmacología , Apoptosis/efectos de los fármacos , Diabetes Mellitus Experimental/tratamiento farmacológico , Transportador de Glucosa de Tipo 4/fisiología , Hipoglucemiantes/farmacología , Resistencia a la Insulina , Páncreas/efectos de los fármacos , Animales , Glucemia/análisis , Peso Corporal/efectos de los fármacos , Diabetes Mellitus Experimental/metabolismo , Hemodinámica/efectos de los fármacos , Masculino , Músculo Esquelético/metabolismo , Estrés Oxidativo/efectos de los fármacos , Páncreas/patología , Ratas , Ratas Sprague-Dawley , Estreptozocina , Triglicéridos/sangreRESUMEN
Wound healing is a complex process that includes inflammation, tissue formation, and remodeling. While wound healing is accompanied by inflammatory reactions, chronic inflammation impairs acute wound healing. In this study, we investigated whether anthocyanins from black soybean seed coats could stimulate wound healing while preventing excessive inflammation. At 24h of treatment with anthocyanins, fibroblasts showed a significant increase in migration at 100 microg/mL whereas the migration of keratinocytes increased significantly at 50 and 100 microg/mL compared to control. Treatment of anthocyanins for 48 h significantly stimulated the migration of both human dermal fibroblasts and keratinocytes at 50 and 100 microg/mL concentrations. Treatment of cells with anthocyanins stimulated wound-induced VEGF production in fibroblasts and keratinocytes. However, anthocyanins inhibited ROS accumulation and VEGF production in TNF-alpha-stimulated endothelial cells. Furthermore, treatment of anthocyanins reduced, in a dose-dependent manner, the adhesion of inflammatory monocytes to endothelial cells. Anthocyanins also blocked both the translocation of nuclear factor-kappa B (NF-kappaB) p65 into the nucleus and the phosphorylation of the inhibitory factor kappaBalpha (IkappaBalpha). Thus, treatment with anthocyanins from black soybean seed coats may be a potential therapeutic strategy to promote wound healing and to prevent inflammation in a persistent inflammatory condition.
Asunto(s)
Antocianinas/farmacología , Células Endoteliales/efectos de los fármacos , Fibroblastos/efectos de los fármacos , Glycine max/química , Inflamación/prevención & control , Queratinocitos/efectos de los fármacos , Antocianinas/química , Ensayos de Migración Celular , Células Cultivadas , Humanos , Biología Molecular , Especies Reactivas de Oxígeno , Semillas/química , Factor de Necrosis Tumoral alfa/farmacología , Factor A de Crecimiento Endotelial Vascular/metabolismoRESUMEN
Adhesion molecules have a key role in pathological inflammation. Thus, we investigated the effect of anthocyanins on the induction of intercellular adhesion molecule-1 (ICAM-1) and vascular cell adhesion molecule-1 (VCAM-1) by TNF-alpha and the possible molecular mechanisms by which anthocyanins differentially regulate ICAM-1 and VCAM-1 expression. Stimulation of cells with TNF-alpha increased ICAM-1 and VCAM-1 expression, and pretreatment with anthocyanins inhibited VCAM-1 expression, but not ICAM-1 expression. We found that IRF-1 and GATAs, especially GATA-4 and -6, were involved in the TNF-alpha-mediated expression of VCAM-1 but not ICAM-1, and anthocyanins decreased nuclear levels of GATA-4 and GATA-6 as well as IRF-1. Moreover, pretreatment with a Jak/STAT inhibitor decreased TNF-alpha-induced VCAM-1 expression and nuclear GATA-4, GATA-6, and IRF-1 levels. Furthermore, anthocyanins efficiently inhibited the phosphorylation of STAT-3. This suggests that anthocyanins differentially regulate TNF-alpha-mediated expression of VCAM-1 and ICAM-1 through modulation of the GATA and IRF-1 binding activity via Jak/STAT-3 activation.
Asunto(s)
Antocianinas/farmacología , Regulación hacia Abajo , Factores de Transcripción GATA/metabolismo , Glycine max/química , Molécula 1 de Adhesión Intercelular/genética , Factor 1 Regulador del Interferón/metabolismo , Factor de Necrosis Tumoral alfa/metabolismo , Molécula 1 de Adhesión Celular Vascular/genética , Células Cultivadas , Endotelio Vascular/efectos de los fármacos , Endotelio Vascular/metabolismo , Factores de Transcripción GATA/genética , Regulación de la Expresión Génica/efectos de los fármacos , Humanos , Molécula 1 de Adhesión Intercelular/metabolismo , Factor 1 Regulador del Interferón/genética , Extractos Vegetales/farmacología , Factor de Necrosis Tumoral alfa/genética , Molécula 1 de Adhesión Celular Vascular/metabolismoRESUMEN
Our previous study suggested that OsBWMK1, a gene which encodes a member of the rice MAP kinase family, generates transcript variants which show distinct expression patterns in response to environmental stresses. The transcript variants are generated by alternative splicing and by use of alternative promoters. To test whether the two alternative promoters, pOsBWMK1L (promoter for the OsBWMK1L splice variant) and pOsBWMK1S (promoter for the OsBWMK1S splice variant), are biologically functional, we analyzed transgenic plants expressing GUS fusion constructs for each promoter. Both pOsBWMK1L and pOsBWMK1S are biologically active, although the activity of pOsBWMK1S is lower than that of pOsBWMK1L. Histochemical analysis revealed that pOsBWMK1L is constitutively active in most tissues at various developmental stages in rice and Arabidopsis, whereas pOsBWMK1S activity is spatially and temporally restricted. Furthermore, the expression of pOsBWMK1S::GUS was upregulated in response to hydrogen peroxide, a plant defense signaling molecule, in both plant species. These results suggest that the differential expression of OsBWMK1 splice variants is the result of alternative promoter usage and, moreover, that the mechanisms controlling OsBWMK1 gene expression are conserved in both monocot and dicot plants.
Asunto(s)
Proteínas Quinasas Activadas por Mitógenos/genética , Oryza/enzimología , Oryza/genética , Proteínas de Plantas/genética , Empalme Alternativo , Secuencia de Bases , Western Blotting , Regulación de la Expresión Génica , Variación Genética , Proteínas Quinasas Activadas por Mitógenos/biosíntesis , Proteínas de Plantas/biosíntesis , Regiones Promotoras Genéticas , Isoformas de ProteínasRESUMEN
Ultraviolet (UV) radiation can cause inflammatory changes and may further contribute to skin carcinogenesis. Anthocyanins are known to be powerful antioxidants that help protect plants from UV damage. Recently, we isolated anthocyanins from black soybean [Glycine max (L.) Merr] seed coats. Thus, we investigated the protective effect of anthocyanins from black soybean seed coats on UVB radiation-induced inflammatory responses and the molecular mechanism responsible for regulation of apoptosis and inflammatory responses. Anthocyanins inhibited UVB-induced cylooxygenase-2 (COX-2) and PGE 2 production through a nuclear factor-kappaB-dependent pathway and regulation of the PI3 kinase/Akt pathway activated by UVB in a human keratinocyte cell line, HaCaT. Topical application of anthocyanins prior to UVB irradiation of hairless mice also inhibited induction of COX-2 and PGE 2. In conclusion, it is suggested that anthocyanins from the seed coat of black soybeans can be used as a useful drug to modulate oxidative disorders including UVB-induced inflammation.
Asunto(s)
Antocianinas/farmacología , Ciclooxigenasa 2/genética , Dinoprostona/antagonistas & inhibidores , Glycine max/química , FN-kappa B/metabolismo , Rayos Ultravioleta , Animales , Línea Celular , Inhibidores de la Ciclooxigenasa 2/farmacología , Dinoprostona/biosíntesis , Femenino , Expresión Génica/efectos de los fármacos , Expresión Génica/efectos de la radiación , Humanos , Queratinocitos , Ratones , Ratones Pelados , Fosfatidilinositol 3-Quinasas/metabolismo , Semillas/química , TransfecciónRESUMEN
UVB radiation proves to be one of the most relevant environmental risks because of its hazardous effects, such as premature skin aging and especially skin photocarcinogenesis. Anthocyanins, water-soluble pigments present in plants, are known to be powerful antioxidants that help protect plants from UV damage. In this study, we aimed at investigating the protective effect of anthocyanins from black soybean [Glycine max (L.) Merr] seed coats on UVB-induced apoptosis, and furthermore, we investigated the molecular mechanism responsible for regulation of apoptosis in vitro and in vivo. Pretreatment with anthocyanins reduced UVB-induced reactive oxygen species levels and inhibited UVB-induced apoptotic cell death through the prevention of caspase-3 pathway activation and reduction of proapoptotic Bax protein levels. UVB irradiation induced apoptotic cell death, which was inhibited by topical application of anthocyanins in hairless mice. It is concluded that anthocyanins from the seed coat of black soybeans may be useful compounds to modulate UVB-induced photoaging.