RESUMEN
The extract from Entada phaseoloides was employed as active ingredients of natural origin into cosmetic products, while the components analysis was barely reported. Using LC-DAD-MS/qTOF analysis, eleven compounds (1-11) were proposed or identified from acetone extract of E. phaseoloides leaves (AE). Among them, six phenolic compounds, protocatechuic acid (2), 4-hydroxybenzoic acid (3), luteolin-7-O-ß-d-glucoside (5), cirsimaritin (6), dihydrokaempferol (9), and apigenin (10), were isolated by various chromatographic techniques. Protocatechuic acid (2), epicatechin (4), and kaempferol (11) at a concentration 100 µM increased the HaCaT cells viability of the UVB-irradiated cell without any cytotoxicity effect and reduced the expression of COX-2 and iNOS inflammation gene. Moreover, compounds 2 and 4 could have potent effects on cell migration during wound closure. These results suggest that compounds 2, 4, and 11 from AE have anti-photoaging properties and could be employed in pharmaceutical and cosmeceutical products.
Asunto(s)
Fabaceae/química , Queratinocitos/efectos de los fármacos , Fenoles/farmacología , Extractos Vegetales/química , Protectores contra Radiación/farmacología , Acetona/química , Línea Celular , Movimiento Celular/efectos de los fármacos , Movimiento Celular/efectos de la radiación , Supervivencia Celular/efectos de los fármacos , Supervivencia Celular/efectos de la radiación , Ciclooxigenasa 2/genética , Regulación Enzimológica de la Expresión Génica/efectos de los fármacos , Regulación Enzimológica de la Expresión Génica/efectos de la radiación , Queratinocitos/efectos de la radiación , Óxido Nítrico Sintasa de Tipo II/genética , Fenoles/química , Protectores contra Radiación/química , Piel/citología , Rayos UltravioletaRESUMEN
BACKGROUND: Cerebral ischemia, a common cerebrovascular disease, is one of the great threats to human health and new targets for stroke therapy are needed. The transcriptional activity in the cell is regulated by epigenetic processes such as DNA methylation/demethylation, acetylation/deacetylation, histone methylation, etc. Changes in DNA methylation after ischemia can have both neuroprotective and neurotoxic effects depending on the degree of ischemia damage, the time elapsed after injury, and the site of methylation. METHODS: In this study, we investigated the changes in the expression and intracellular localization of DNA methyltransferase DNMT1, histone methyltransferases SUV39H1, and G9a in penumbra neurons and astrocytes at 4 and 24 h after stroke in the rat cerebral cortex using photothrombotic stroke (PTS) model. Methods of immunofluorescence microscopy analysis, apoptosis analysis, and immunoblotting were used. Additionally, we have studied the effect of DNMT1 and G9a inhibitors on the volume of PTS-induced infarction and apoptosis of penumbra cells in the cortex of mice after PTS. RESULTS: This study has shown that the level of DNMT1 increased in the nuclear and cytoplasmic fractions of the penumbra tissue at 24 h after PTS. Inhibition of DNMT1 by 5-aza-2'-deoxycytidine protected cells of PTS-induced penumbra from apoptosis. An increase in the level of SUV39H1 in the penumbra was found at 24 h after PTS and G9a was overexpressed at 4 and 24 h after PTS. G9a inhibitors A-366 and BIX01294 protected penumbra cells from apoptosis and reduced the volume of PTS-induced cerebral infarction. CONCLUSION: Thus, the data obtained show that DNA methyltransferase DNMT1 and histone methyltransferase G9a can be potential protein targets in ischemic penumbra cells, and their inhibitors are potential neuroprotective agents capable of protecting penumbra cells from postischemic damage to the cerebral cortex.
Asunto(s)
ADN (Citosina-5-)-Metiltransferasa 1/genética , N-Metiltransferasa de Histona-Lisina/genética , Metiltransferasas/genética , Neuronas/metabolismo , Proteínas Represoras/genética , Accidente Cerebrovascular/genética , Animales , Astrocitos/metabolismo , Astrocitos/efectos de la radiación , Corteza Cerebral/metabolismo , Corteza Cerebral/efectos de la radiación , Metilación de ADN/efectos de la radiación , Modelos Animales de Enfermedad , Regulación Enzimológica de la Expresión Génica/efectos de la radiación , Humanos , Luz , Ratones , Neuronas/patología , Neuronas/efectos de la radiación , Ratas , Accidente Cerebrovascular/patología , Accidente Cerebrovascular/terapiaRESUMEN
An electromagnetic field (EMF) may have effects on female reproduction. This study was conducted to determine whether EMF [50 and 120â¯Hz, 2 and 4â¯h of incubation in the presence or absence of progesterone (P4, 10-5 M)] affects androgen synthesis and release from the pig endometrium. Endometrial slices were collected from pigs (n = 5) during the fetal peri-implantation period (i.e., days 15-16 of gestation) and treated in vitro with EMF. The selected endometrial slices were treated with P4 to determine whether this hormone has effects on protection of the tissue from EMF radiation. The CYP17A1 and HSD3B1 mRNA transcript abundance, steroid 17αhydroxylase/17, 20-lyase (cytochrome P450c17) and hydroxyΔ5steroid dehydrogenase/3ß and steroidΔisomerase (3ßHSD) protein abundance were examined using Real-Time PCR and Western Blot procedures, respectively. In media collected after incubation, the concentrations of androstenedione (A4) and testosterone (T) were quantified used a RIA. When P4 was added to the culture medium, EMF radiation had suppressive effects on endometrial T release after 2 and 4â¯h of incubation when the EMF treatment was occurring and increased A4 release after 4â¯h of incubation with EMF at 120â¯Hz. When there was no inclusion of P4, release of A4 was decreased after 2â¯h of EMF treatment at 120â¯Hz and after 4â¯h of EMF treatment at 50 and 120â¯Hz. Progesterone did not have functions that protected the pig endometrium against EMF radiation during the fetal peri-implantation period.
Asunto(s)
Campos Electromagnéticos/efectos adversos , Implantación del Embrión/efectos de la radiación , Endometrio/efectos de la radiación , Porcinos/fisiología , Testosterona/metabolismo , Animales , Femenino , Regulación Enzimológica de la Expresión Génica/efectos de la radiación , Embarazo , Progesterona/metabolismo , Progesterona Reductasa/genética , Progesterona Reductasa/metabolismo , Esteroide 17-alfa-Hidroxilasa/genética , Esteroide 17-alfa-Hidroxilasa/metabolismo , Esteroide Isomerasas/genética , Esteroide Isomerasas/metabolismoRESUMEN
PURPOSE: Safety concerns about the effects of long-term extremely low-frequency electromagnetic field (ELF-EMF) exposure on human health have been raised. To explore the effects of continuous exposure to ELF-EMF on organisms for multiple generations, we selected Caenorhabditis elegans as a model organism and conducted long-term continuous exposure studies for multiple generations under 20 °C, 50 Hz, and 3 mT ELF-EMF. MATERIALS AND METHODS: Each generation of worms was treated with ELF-EMF from the egg in the same environment. After long-term exposure to ELF-EMF, the body length of the worms was detected, and 15th generation adult worms were selected as the research object. The ATP level and ATPase were detected, and the expression levels of genes encoding ATP synthase (r53.4, hpo-18, atp-5, unc-32, atp-3) were detected by RT-PCR. In worm's antioxidant system, the level of reactive oxygen species (ROS) was detected by dichlorofluorescein staining, and the total antioxidant capacity (T-AOC), superoxide dismutase (SOD) and catalase (CAT) activity were investigated. The expression of genes encoding superoxide dismutase (sod-1, sod-2, sod-3) was detected in adult (60 h) worms of the fifteenth generation (F15). RESULTS: These results showed that the body length of F15 worms increased significantly, ATP content increased significantly, ATP synthase activity was significantly enhanced, and the expression levels of the r53.4, hpo-18, atp-5, and atp-3 genes encoding ATPase were significantly upregulated in F15 worms. In addition, SOD activity increased significantly, and the expression levels of the sod-1, sod-2, and sod-3 genes encoding SOD were also significantly upregulated in F15 worms. CONCLUSIONS: These results indicated that continuous exposure to 50 Hz, 3 mT ELF-EMF for multiple generations can increase the body length of worms, induce the synthesis of ATP and enhance the antioxidant capacity of worms.
Asunto(s)
Adenosina Trifosfato/metabolismo , Antioxidantes/metabolismo , Caenorhabditis elegans/metabolismo , Caenorhabditis elegans/efectos de la radiación , Campos Electromagnéticos/efectos adversos , Adenosina Trifosfatasas/metabolismo , Animales , Regulación Enzimológica de la Expresión Génica/efectos de la radiación , Superóxido Dismutasa/metabolismo , Factores de TiempoRESUMEN
An electromagnetic field (EMF) has been found to affect reproductive processes in females. The aim of this study was to determine the effect of low, non-ionizing EMF radiation on the steroidogenic activity of myometrium collected from pigs during the fetal peri-implantation period. Myometrial slices were treated with an EMF (50 and 120â¯Hz, 2 and 4â¯h of incubation) and examined for the aromatase cytochrome P450 17α-hydroxylase/C17-20lyase (CYP17A1) and 3ß-hydroxysteroid dehydrogenase/Δ5-Δ4 isomerase (HSD3B1) mRNA transcript abundance, cytochrome P450c17 and 3ßHSD protein abundance and the secretion of androstenedione (A4) and testosterone (T). To determine whether progesterone (P4) functions as a protectant from EMF radiation, the selected slices were treated with P4. In slices incubated without P4, EMF at 50â¯Hz altered cytochrome P450c17 protein abundance (4â¯h), HSD3B1 mRNA transcript abundance (4â¯h) and A4 release (2â¯h) as well as T release (2â¯h) in P4-treated slices. The EMF at 120â¯Hz in non P4-treated slices altered A4 release (2 and 4â¯h) whereas in P4-treated slices altered CYP17A1 mRNA transcript abundance (4â¯h), 3ßHSD protein abundance (4â¯h), A4 (4â¯h) and T release (2â¯h). In conclusion, EMF radiation in the myometrium collected during the peri-implantation period alters the CYP17A1 and HSD3B1 mRNA transcript and encoded protein abundance, and androgen release due to the time of treatment and P4 presence or absence. The P4 did not function directly as an obvious protector against EMF radiation in the myometrium of pigs during the fetal peri-implantation period.
Asunto(s)
Andrógenos/biosíntesis , Campos Electromagnéticos/efectos adversos , Miometrio/efectos de la radiación , Porcinos/metabolismo , Andrógenos/metabolismo , Animales , Femenino , Regulación Enzimológica de la Expresión Génica/efectos de los fármacos , Regulación Enzimológica de la Expresión Génica/efectos de la radiación , Miometrio/metabolismo , Embarazo , Progesterona/farmacología , Progesterona Reductasa/genética , Progesterona Reductasa/metabolismo , Esteroide 17-alfa-Hidroxilasa/genética , Esteroide 17-alfa-Hidroxilasa/metabolismo , Esteroide Isomerasas/genética , Esteroide Isomerasas/metabolismoRESUMEN
Skin aging is primarily associated with the alterations in dermal extracellular matrix, in particular a decrease in collagen type-1 content. Recent studies have shown that collagen-degrading matrix metalloproteinase (MMP-1) is produced by fibroblasts in response to chronoaging, which in human dermal fibroblasts leads to the release of proinflammatory cytokines. Past studies showed that anti-inflammatory capabilities could be induced via non-chemical means. One of these methods makes use of ultra-weak fractal electromagnetic (uwf-EM) signals. Such ultra-/very-low frequency (U/VLF) signals (few nT in intensity and within 0.5-30 kHz) interact with aqueous solutions in living systems. The fractal nature of such EM-signals relates to the self-similar property by which a "cut-out" and magnified piece of this signal reveals again the original. Thus, the aim of this study is twofold, to i) investigate the extent of this modulating effect using Human Dermal Fibroblasts (HDF)-cells, and ii) analyse molecular rejuvenation markers therein. We could demonstrate that a 10 min uwf-EM exposure (prior to incubation) increases type-1 collagen and modulates elastin in human fibroblasts cultured up to 96 h, while at the same time reduces IL-6, TNF-α and MMP-1 (the later three being statistically significant). Such up- respectively down-regulation of corresponding genes are strong indicators of an EM-induced hormetic effect that influences the epigenomic landscape of HDFs. In the Appendix, we present, in the framework of Quantum Field Theory (QFT), water as a biphasic liquid and how its coherent fraction can be affected by uwf-EM signals while at the same time resolving the "kT paradox".
Asunto(s)
Campos Electromagnéticos/efectos adversos , Fibroblastos/metabolismo , Fibroblastos/efectos de la radiación , Fractales , Biomarcadores/metabolismo , Supervivencia Celular/efectos de la radiación , Colágeno Tipo I/genética , Citocinas/metabolismo , Dermis/metabolismo , Dermis/efectos de la radiación , Elastina/genética , Fibroblastos/citología , Regulación Enzimológica de la Expresión Génica/efectos de la radiación , Humanos , Metaloproteinasa 1 de la Matriz/genéticaRESUMEN
In this study, we investigated the effects of specific low-frequency electromagnetic field sequences on U937 cells, an in vitro model of human monocyte/macrophage differentiation. U937 cells were exposed to electromagnetic stimulation by means of the SynthéXer system using two similar sequences, XR-BC31 and XR-BC31/F. Each sequence was a time series of 29 wave segments, equal to a total duration of 77 min. Here, we report that exposure (4 d, once a day) of U937 cells to the XR-BC31 setting, but not to the XR-BC31/F, resulted in increased expression of the histone demethylase KDM6B along with a global reduction in histone H3 lysine 27 tri-methylation (H3K27me3). Furthermore, exposure to the XR-BC31 sequence induced differentiation of U937 cells towards a macrophage-like phenotype displaying a KDM6B dependent increase in expression and secretion of the anti-inflammatory interleukins (ILs), IL-10 and IL-4. Importantly, all the observed changes were highly dependent on the nature of the sequence. Our results open a new way of interpretation for the effects of low-frequency electromagnetic fields observed in vivo. Indeed, it is conceivable that a specific low-frequency electromagnetic fields treatment may cause the reprogramming of H3K27me3 and cell differentiation.
Asunto(s)
Campos Electromagnéticos , Regulación Enzimológica de la Expresión Génica/efectos de la radiación , Histona Demetilasas con Dominio de Jumonji/genética , Ciclo Celular/efectos de la radiación , Histonas/metabolismo , Humanos , Interleucina-10/metabolismo , Interleucina-4/metabolismo , Metilación/efectos de la radiación , Fenotipo , Células U937RESUMEN
Catechins and theaflavins are important metabolites contributing to tea function and quality. Catechins are known to transform into theaflavins during the tea manufacturing process, but the same transformation in preharvest tea leaves is unknown. Herein, we determined that shade treatment (dark), an agronomic practise widely used in tea cultivation, reduced the contents of most catechins, but increased the theaflavin contents, in preharvest tea leaves (cv. Yinghong No.9). This was attributed to the activation of polyphenoloxidase (PPO) activity in darkness. Furthermore, CsPPO3 was highly expressed under darkness, and thus CsPPO3 had been cloned, sequenced, and characterization. The CsPPO3 recombinant protein exhibited PPO function. Furthermore, shade treatment also reduced the catechin contents and increased the theaflavin contents in Yabukita and Hoshinomidori, suggesting that this phenomenon might not be specific to certain tea cultivars. This information will aid in understanding of theaflavin formation and its response to environmental factors at the preharvest tea stage.
Asunto(s)
Biflavonoides/metabolismo , Camellia sinensis/enzimología , Catequina/metabolismo , Catecol Oxidasa/metabolismo , Luz , Hojas de la Planta/enzimología , Biflavonoides/química , Camellia sinensis/química , Catequina/química , Catecol Oxidasa/genética , Regulación Enzimológica de la Expresión Génica/efectos de la radiación , Regulación de la Expresión Génica de las Plantas/efectos de la radiación , Estructura Molecular , Hojas de la Planta/química , Regulación hacia ArribaRESUMEN
Regulation of plant cell wall degradation is of utmost importance for understanding the carbon cycle in nature, but also to improve industrial processes aimed at enzyme production for next generation biofuels. Thereby, the transcription factor networks in different fungi show conservation as well as striking differences, particularly between Trichoderma reesei and Neurospora crassa. Here, we aimed to gain insight into the function of the transcription factors CLR1 and CLR2 in T. reesei, which are crucial for cellulase gene expression in N. crassa. We studied impacts on gene regulation with cellulose, xylan, pectin and chitin, growth on 95 different carbon sources as well as an involvement in regulation of secondary metabolism or development. We found that CLR1 is present in the genome of T. reesei and other Trichoderma spp., albeit with considerably lower homology compared to other ascomycetes. CLR1 and CLR2 regulate pectinase transcript levels upon growth on pectin, no major function was detected on chitin. CLR1 and CLR2 form a positive feedback cycle on xylan and were found to be responsible for balancing co-regulation of xylanase genes in light and darkness with distinct and in part opposite regulatory effects of up to 8fold difference. Our data suggest that CLR1 and CLR2 have evolved differently in T. reesei compared to other fungi. We propose a model in which their main function is in adjustment of regulation of xylanase gene expression to different light conditions and to balance transcript levels of genes involved in plant cell wall degradation according to their individual relevance for this process.
Asunto(s)
Endo-1,4-beta Xilanasas/genética , Hypocreales/genética , Luz , Poligalacturonasa/genética , Factores de Transcripción/genética , Pared Celular/metabolismo , Proteínas de Unión al ADN/genética , Proteínas de Unión al ADN/metabolismo , Endo-1,4-beta Xilanasas/metabolismo , Proteínas Fúngicas/genética , Proteínas Fúngicas/metabolismo , Regulación Enzimológica de la Expresión Génica/efectos de la radiación , Regulación Fúngica de la Expresión Génica/efectos de la radiación , Genes Fúngicos , Hypocreales/metabolismo , Hypocreales/efectos de la radiación , Mutación , Filogenia , Poligalacturonasa/metabolismo , Metabolismo Secundario/genética , Factores de Transcripción/metabolismo , Xilanos/metabolismoRESUMEN
Light intensity plays an important role in the regulation of growth, behavior, reproduction, and welfare of avian species. Light intensity preference behavior has been suggested to be involved in welfare of birds. This study aims to investigate the effects of different light intensity and dual light intensity choice (DLIC) lighting program on plasma corticosterone (CORT), and tryptophan hydroxylase 2 (TPH2, the rate-limiting enzyme of serotonin biosynthesis) and tyrosine hydroxylase (TH, the rate-limiting enzyme of dopamine biosynthesis) gene expression in the brainstem of male chickens. Day old broilers were housed in two commercial houses, and placed in 24 pens. All the treatment groups were provided with 23â¯h light (L) /1â¯h dark (D) and 30â¯lx (lx) light intensity during the first week and then 18L:6D (10â¯lx) from day 7 to 14. Blood and brain were sampled at 14â¯days of age (10â¯lx) before the onset of light treatments. On day 15, four treatments (2, 10, 20, and 100â¯lx), and DLIC treatment (2/20â¯lx) were initiated. Samples were collected on days 15, 16, 17, 30 and 41. TPH2 expression in the dorsal raphe nucleus (DRN) and caudal raphe nucleus (CRN) of brainstem, and TPH2 and TH expression in ventral tegmental areas (VTN) of the midbrain were determined by qPCR. Results showed that bright light and DLIC lighting program temporarily attenuated plasma CORT, suggesting the short-term stress attenuating effect of bright light and DLIC lighting program. Differential TPH2 expression in the DRN and CRN observed in the DLIC birds indicate a significant effect of DLIC lighting program on the serotonergic activity in the avian brainstem. At the 41â¯days of age, the significant downregulation of TPH2 and TH expression occurred in the VTA of DLIC treated birds compared to the other group of birds. Taken together, temporal and spatial regulation of TPH2 and TH expression by DLIC lighting program indicate that compensatory regulation of serotonergic and dopaminergic activities might be involved in the light intensity preference behavior of birds, suggesting a possible beneficial effect of the DLIC lighting program on broiler welfare.
Asunto(s)
Pollos/sangre , Pollos/metabolismo , Corticosterona/sangre , Dopamina/metabolismo , Luz , Serotonina/metabolismo , Animales , Tronco Encefálico/metabolismo , Tronco Encefálico/efectos de la radiación , Pollos/crecimiento & desarrollo , Regulación Enzimológica de la Expresión Génica/efectos de la radiación , Masculino , Núcleos del Rafe/metabolismo , Núcleos del Rafe/efectos de la radiación , Triptófano Hidroxilasa/genética , Triptófano Hidroxilasa/metabolismo , Tirosina 3-Monooxigenasa/genética , Tirosina 3-Monooxigenasa/metabolismo , Área Tegmental Ventral/metabolismoRESUMEN
Breast cancer (BC) is the most common tumour in women and one of the most important causes of cancer death worldwide. Radiation therapy (RT) is widely used for BC treatment. Some proteins have been identified as prognostic factors for BC (Ki67, p53, E-cadherin, HER2). In the last years, it has been shown that variations in the expression of MMPs and TIMPs may contribute to the development of BC. The aim of this pilot work was to study the effects of RT on different MMPs (-1, -2, -3, -7, -8, -9, -10, -12 and -13) and TIMPs (-1 to -4), as well as their relationship with other variables related to patient characteristics and tumour biology. A group of 20 BC patients treated with RT were recruited. MMP and TIMP serum levels were analysed by immunoassay before, during and after RT. Our pilot study showed a slight increase in the levels of most MMP and TIMP with RT. However, RT produced a significantly decrease in TIMP-1 and TIMP-3 levels. Significant correlations were found between MMP-3 and TIMP-4 levels, and some of the variables studied related to patient characteristics and tumour biology. Moreover, MMP-9 and TIMP-3 levels could be predictive of RT toxicity. For this reason, MMP-3, MMP-9, TIMP-3 and TIMP-4 could be used as potential prognostic and predictive biomarkers for BC patients treated with RT.
Asunto(s)
Neoplasias de la Mama/patología , Regulación Enzimológica de la Expresión Génica/efectos de la radiación , Metaloproteinasas de la Matriz/sangre , Radioterapia/métodos , Inhibidores Tisulares de Metaloproteinasas/sangre , Neoplasias de la Mama/sangre , Neoplasias de la Mama/enzimología , Neoplasias de la Mama/radioterapia , Femenino , Humanos , Persona de Mediana Edad , Proyectos Piloto , PronósticoRESUMEN
Radiotherapy (RT) is currently one of the leading treatments for various cancers; however, it may cause damage to healthy tissue, with both short-term and long-term side effects. Severe radiation-induced normal tissue damage (RINTD) frequently has a significant influence on the progress of RT and the survival and prognosis of patients. The redox system has been shown to play an important role in the early and late effects of RINTD. Reactive oxygen species (ROS) and reactive nitrogen species (RNS) are the main sources of RINTD. The free radicals produced by irradiation can upregulate several enzymes including nicotinamide adenine dinucleotide phosphate oxidase (NADPH oxidase), lipoxygenases (LOXs), nitric oxide synthase (NOS), and cyclooxygenases (COXs). These enzymes are expressed in distinct ways in various cells, tissues, and organs and participate in the RINTD process through different regulatory mechanisms. In recent years, several studies have demonstrated that epigenetic modulators play an important role in the RINTD process. Epigenetic modifications primarily contain noncoding RNA regulation, histone modifications, and DNA methylation. In this article, we will review the role of oxidative stress and epigenetic mechanisms in radiation damage, and explore possible prophylactic and therapeutic strategies for RINTD.
Asunto(s)
Epigénesis Genética/efectos de la radiación , Regulación Enzimológica de la Expresión Génica/efectos de la radiación , Regulación Neoplásica de la Expresión Génica/efectos de la radiación , Neoplasias/enzimología , Estrés Oxidativo/efectos de la radiación , Oxidorreductasas/biosíntesis , Traumatismos por Radiación/enzimología , Radioterapia/efectos adversos , Animales , Humanos , Neoplasias/patología , Neoplasias/radioterapia , Traumatismos por Radiación/patología , Especies Reactivas de Oxígeno/metabolismoRESUMEN
Radiation induced damage to normal cells is a major shortcoming of conventional radiotherapy, which necessitates the development of novel radio-protective drugs. An ideal radio-modulator would protect normal cells while having cytotoxic effects on cancer cells. Plumbagin is a potent anti-tumour agent and has been shown to sensitize tumour cells to radiation-induced damage. In the present study, we have evaluated the radio-protective potential of plumbagin and found that it protected normal lymphocytes against radiation-induced apoptosis, but did not protect cancer cells against radiation. Plumbagin offered radioprotection even when it was added to cells after irradiation. The ability of only thiol based antioxidants to abrogate the radio-protective effects of plumbagin suggested a pivotal role of thiol groups in the radio-protective activity of plumbagin. Further, protein interaction network (PIN) analysis was used to predict the molecular targets of plumbagin. Based on the inputs from plumbagin's PIN and in light of its well-documented ability to modulate thiol groups, we proposed that plumbagin may act via modulation of caspase enzyme which harbours a critical catalytic cysteine. Indeed, plumbagin suppressed radiation-induced increase in homogenous caspase and caspase-3 activity in lymphocytes. Plumbagin also inhibited the activity of recombinant caspase-3 and mass spectrometric analysis revealed that plumbagin covalently interacts with caspase-3. Further, the in vivo radioprotective efficacy of plumbagin (single dose of 2mg/kg body weight) was demonstrated by its ability to rescue mice against radiation (7.5â¯Gy; Whole Body Irradiation) induced mortality. These results indicate that plumbagin prevents radiation induced apoptosis specifically in normal cells by inhibition of caspase-3 activity.
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Caspasa 3/metabolismo , Rayos gamma/efectos adversos , Regulación Enzimológica de la Expresión Génica/efectos de los fármacos , Neoplasias Pulmonares/enzimología , Linfocitos/enzimología , Naftoquinonas/farmacología , Protectores contra Radiación/farmacología , Animales , Antineoplásicos Fitogénicos/farmacología , Apoptosis , Caspasa 3/genética , Proliferación Celular , Regulación Enzimológica de la Expresión Génica/efectos de la radiación , Humanos , Neoplasias Pulmonares/tratamiento farmacológico , Neoplasias Pulmonares/radioterapia , Linfocitos/efectos de los fármacos , Linfocitos/efectos de la radiación , Ratones , Oxidación-Reducción , FosforilaciónRESUMEN
Zeaxanthin is an important pigment in the photo-protection mechanism of microalgae. However, zeaxanthin epoxidase, an enzyme involved in the accumulation and conversion of zeaxanthin, has not been extensively studied in microalgae. In this work, we report the expression pattern of zeaxanthin epoxidase in Dunaliella tertiolecta (DtZEP) at different light and diverse salinity conditions. To confirm the responsiveness to light conditions, the ZEP expression pattern was investigated in photoperiodic (16 h of light and 8 h of dark) and continuous (24 h of light and 0 h of dark) light conditions. mRNA expression levels in photoperiodic conditions fluctuated along with the light/dark cycle, whereas those in continuous light remained unchanged. In varying salinity conditions, the highest mRNA and protein levels were detected in cells cultured in 1.5 M NaCl, and ZEP expression levels in cells shifted from 0.6 M NaCl to 1.5 M NaCl increased gradually. These results show that mRNA expression of DtZEP responds rapidly to the light/dark cycle or increased salinity, whereas changes in protein synthesis do not occur within a short period. Taken together, we show that DtZEP gene expression responds rapidly to light irradiation and hyperosmotic stress. In addition, ZEP expression patterns in light or salinity conditions are similar to those of higher plants, even though the habitat of D. tertiolecta is different.
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Chlorophyceae/enzimología , Regulación Enzimológica de la Expresión Génica , Regulación de la Expresión Génica de las Plantas , Microalgas/enzimología , Oxidorreductasas/genética , Vías Biosintéticas/genética , Carotenoides/metabolismo , Chlorophyceae/genética , Chlorophyceae/metabolismo , Regulación Enzimológica de la Expresión Génica/efectos de los fármacos , Regulación Enzimológica de la Expresión Génica/efectos de la radiación , Regulación de la Expresión Génica de las Plantas/efectos de los fármacos , Regulación de la Expresión Génica de las Plantas/efectos de la radiación , Luz , Microalgas/genética , Microalgas/metabolismo , Oxidorreductasas/metabolismo , Fotoperiodo , ARN Mensajero/genética , ARN Mensajero/metabolismo , Salinidad , Cloruro de Sodio/farmacologíaRESUMEN
Radiation-induced pulmonary fibrosis (RIPF) is a chronic, progressive complication of therapeutic irradiation of the thorax. It has been suggested that senescence of type II pneumocytes (AECIIs), an alveolar stem cell, plays a role in the development of RIPF through loss of replicative reserve and via senescent AECII-driven release of proinflammatory and profibrotic cytokines. Within this context, we hypothesized that arachidonate 12-lipoxygenase (12-LOX) is a critical mediator of AECII senescence and RIPF. Treatment of wild-type AECIIs with 12S-hydroxyeicosateraenoic acid (12S-HETE), a downstream product of 12-LOX, was sufficient to induce senescence in a NADPH oxidase 4 (NOX4)-dependent manner. Mice deficient in 12-LOX exhibited reduced AECII senescence, pulmonary collagen accumulation and accumulation of alternatively activated (M2) macrophages after thoracic irradiation (5 × 6 Gy) compared to wild-type mice. Conditioned media from irradiated or 12S-HETE-treated primary pneumocytes contained elevated levels of IL-4 and IL-13 compared to untreated pneumocytes. Primary macrophages treated with conditioned media from irradiated AECII demonstrated preferential M2 type polarization when AECIIs were derived from wild-type mice compared to 12-LOX-deficient mice. Together, these data identified 12-LOX as a critical component of RIPF and a therapeutic target for radiation-induced lung injury.
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Células Epiteliales Alveolares/patología , Araquidonato 12-Lipooxigenasa/metabolismo , Senescencia Celular/efectos de la radiación , Macrófagos/efectos de la radiación , Neumonitis por Radiación/enzimología , Células Epiteliales Alveolares/efectos de la radiación , Animales , Araquidonato 12-Lipooxigenasa/genética , Femenino , Regulación Enzimológica de la Expresión Génica/efectos de la radiación , Interleucina-13/biosíntesis , Interleucina-4/biosíntesis , Macrófagos/citología , Macrófagos/metabolismo , Ratones , Ratones Endogámicos C57BL , Neumonitis por Radiación/genética , Neumonitis por Radiación/inmunología , Neumonitis por Radiación/patologíaRESUMEN
Protein kinase CK2 is a ubiquitously expressed kinase in eukaryotes, which is known to phosphorylate many protein substrates. Because CK2 is involved in the regulation of various signaling pathways, we wondered whether CK2 participated in the regulation of ionizing radiation (IR) induced biological process. In this study, we investigated the effect of IR on the subcellular localization and kinase activity in human non-small cell lung cancer (NSCLC) cells. Immunofluorescent results showed that CK2 subunits shuttle into the nucleus mostly beginning 1 h after IR and lasting more than 6 h. We also conducted in vitro kinase assay and observed an increase in CK2 kinase activity at 6 h after IR. Furthermore, an increase in S phase was observed at 6 h after IR. Colony formation assay results demonstrated that CK2 inhibitor CX-4945 significantly enhanced the effect of irradiation in NSCLC cells. These results indicated that CK2 may be implicated in the regulation of IR-induced biological process.
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Carcinoma de Pulmón de Células no Pequeñas/enzimología , Regulación Enzimológica de la Expresión Génica/efectos de la radiación , Regulación Neoplásica de la Expresión Génica/efectos de la radiación , Neoplasias Pulmonares/enzimología , Radiación Ionizante , Quinasa de la Caseína II/metabolismo , Ciclo Celular , Línea Celular Tumoral , Núcleo Celular/metabolismo , Supervivencia Celular , Citoplasma/metabolismo , Citosol/metabolismo , Genotipo , Humanos , Naftiridinas/farmacología , Fenazinas , Fosforilación , Transducción de SeñalRESUMEN
In this study, we investigated the protective effects of a peptide (YGDEY, Tyr-Gly-Asp-Glu-Tyr) isolated from tilapia skin gelatin hydrolysates (TGHs), against UVB-induced photoaging in human keratinocytes (HaCaT) cells. Results showed that YGDEY significantly decreased levels of intracellular reactive oxygen species (ROS), increased antioxidant factors (Superoxide Dismutase, SOD and Glutathione, GSH) expression and maintained balance between GSH and GSSG in HaCaT cells. Comet assay shows that YGDEY can protect DNA from oxidative damage. Furthermore, it significantly inhibited MMP-1 (collagenase) and MMP-9 (gelatinase) expression and increased Type I procollagen production. In addition, the molecular docking study showed that YGDEY may form active sites with MMP-1 and MMP-9. Moreover, Western blot analysis was utilized to measure the protein levels of UVB-induced mitogen-activated protein kinase (MAPK) and nuclear factor-kappa B (NF-κB) signaling pathways. Therefore, these results suggested that YGDEY has a therapeutic effectiveness in prevention of UVB-induced cellular damage, and it is a candidate worthy of being developed as a potential natural antioxidant and food additive.
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Gelatina/química , Metaloproteinasa 1 de la Matriz/metabolismo , Metaloproteinasa 9 de la Matriz/metabolismo , Péptidos/farmacología , Envejecimiento de la Piel/efectos de los fármacos , Tilapia , Animales , Línea Celular , Supervivencia Celular/efectos de los fármacos , Supervivencia Celular/efectos de la radiación , Daño del ADN , Regulación Enzimológica de la Expresión Génica/efectos de los fármacos , Regulación Enzimológica de la Expresión Génica/efectos de la radiación , Humanos , Queratinocitos/efectos de los fármacos , Queratinocitos/efectos de la radiación , Metaloproteinasa 1 de la Matriz/genética , Metaloproteinasa 9 de la Matriz/genética , Modelos Moleculares , Oxidación-Reducción , Péptidos/química , Conformación Proteica , Envejecimiento de la Piel/efectos de la radiación , Rayos UltravioletaRESUMEN
Vina-ginsenoside R7 (R7) has been exhibited to engage in multiple pharmacological activities, such as antioxidant and anti-inflammatory activities. However, no photoaging-related studies have been performed on R7. Research is being conducted with the aim of assessing whether treatment with R7 has a protective effect on UVB-induced photoaging skin. Our results show that UVB exposure directly reduces matrix metalloproteinase (MMP) secretion through R7 by restraining the AP-1/MAPK pathway and blocks extracellular matrix (ECM) expression degradation. In addition, R7 improves the expression of transforming growth factor beta 1 (TGF-ß1), and type I procollagen also facilitates the synthesis of collagen by the TGF-ß/Smad signal transduction pathway. Finally, R7 valid blocks nuclear factor-κB (NF-κB) activation and enhances antioxidative stress capacity through activated nuclear factor (erythroid derived 2)-like 2 (Nrf2). In particular, the application of R7 restrains pro-inflammatory cytokines (TNF-α, IL-6, iNOS), which trigger ECM, degrade enzyme production, and suppress vascular endothelial growth factor (VEGF) secretion. In conclusion, R7 may constitute a promising cosmetic ingredient that can protect against skin photodamage resulting from detrimental UVB irradiation.
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Antiinflamatorios/farmacología , Fibroblastos/efectos de los fármacos , Fibroblastos/efectos de la radiación , Ginsenósidos/farmacología , Proteínas Luminiscentes/farmacología , Piel/citología , Rayos Ultravioleta/efectos adversos , Línea Celular , Supervivencia Celular/efectos de los fármacos , Supervivencia Celular/efectos de la radiación , Colágeno Tipo I/metabolismo , Activación Enzimática/efectos de los fármacos , Activación Enzimática/efectos de la radiación , Fibroblastos/citología , Fibroblastos/metabolismo , Regulación Enzimológica de la Expresión Génica/efectos de los fármacos , Regulación Enzimológica de la Expresión Génica/efectos de la radiación , Humanos , Interleucina-6/metabolismo , Espacio Intracelular/efectos de los fármacos , Espacio Intracelular/metabolismo , Espacio Intracelular/efectos de la radiación , Metaloproteinasa 1 de la Matriz/genética , Metaloproteinasa 1 de la Matriz/metabolismo , Metaloproteinasa 3 de la Matriz/genética , Metaloproteinasa 3 de la Matriz/metabolismo , Proteínas Quinasas Activadas por Mitógenos/metabolismo , FN-kappa B/metabolismo , Proteolisis/efectos de los fármacos , Proteolisis/efectos de la radiación , ARN Mensajero/genética , ARN Mensajero/metabolismo , Especies Reactivas de Oxígeno/metabolismo , Transducción de Señal/efectos de los fármacos , Transducción de Señal/efectos de la radiación , Piel/efectos de los fármacos , Piel/efectos de la radiación , Factor de Crecimiento Transformador beta1/metabolismo , Factor A de Crecimiento Endotelial Vascular/biosíntesisRESUMEN
Early ripening apples are usually used for fresh marketing because of short storage life, although they are with high acid and low sugar contents. Understanding the malate metabolism in fleshy fruit and underpinning process during ripening is crucial for particular crop improvement where acidity is a concern for direct consumption or further processing. In this research, a traditional Chinese apple cultivar 'Hongyu', which belongs to early ripening apple cultivar, were freshly harvested at commercial maturity stage (120 Days after full bloom) and used for different storage temperature (4°C, 20°C) and UV-C treatment (following storage at 20°C after treatment). Simple sugars (glucose, sucrose, and fructose) and organic acids (malic, and oxalic) were assessed after 14 d of storage. Compared to fruits stored at 20°C, the malate content in fruits stored at 4°C significantly higher, while it was decreased significantly in UV-C treated fruits stored at 20°C after 14 d of storage. The sugar content was almost similar throughout the UV-C-treated fruits and fruits stored at different temperature. The higher ratios of total sugars to total organic acids in UV-C treated fruits after 14 d suggest that UV-C treatment has the potential to improve the taste of early ripening apple cultivars. Considering the significant difference in malate the samples at 14 d of storage were subjected for RNA-seq analysis. Transcriptome analysis revealed that the phenomena underlying this change were governed by metabolism of malate by the regulation of NADP-malic enzyme (NADP-ME) and phosphoenolpyruvate carboxylase kinase (PEPCK) in apple during postharvest storage. This transcriptome profiling results have specified the transcript regulation of malate metabolism and lead to possible taste improvement without affecting the other fruit quality attributes.
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Almacenamiento de Alimentos , Frutas/crecimiento & desarrollo , Regulación Enzimológica de la Expresión Génica/efectos de la radiación , Regulación de la Expresión Génica de las Plantas/efectos de la radiación , Malato Deshidrogenasa/biosíntesis , Malatos/metabolismo , Malus/crecimiento & desarrollo , Proteínas de Plantas/biosíntesis , Rayos Ultravioleta , Perfilación de la Expresión GénicaRESUMEN
OBJECTIVE: It has been well established that sumoylation acts as an important regulatory mechanism that controls many different cellular processes. We and others have shown that sumoylation plays an indispensable role during mouse eye development. Whether sumoylation is implicated in ocular pathogenesis remains to be further studied. In the present study, we have examined the expression patterns of the de-sumoylation enzymes (SENPs) in the in vitro cataract models induced by glucose oxidase and UVA irradiation. METHODS: Four-week-old C57BL/6J mice were used in our experiments. Lenses were carefully dissected out from mouse eyes and cultured in M199 medium for 12 hours. Transparent lenses (without surgical damage) were selected for experimentation. The lenses were exposed to UVA for 60 min or treated with 20 mU/mL glucose oxidase (GO) to induce cataract formation. The mRNA levels were analyzed with qRT-PCR. The protein levels were determined with western blot analysis and quantitated with Image J. RESULTS: GO treatment and UVA irradiation can induce cataract formation in lens cultured in vitro. GO treatment significantly down-regulated the mRNA levels for SENPs from 50% to 85%; on the other hand, expression of seven SENP proteins under GO treatment appeared in 3 situations: upregulation for SENP1, 2 and 6; downregulation for SENP 5 and 8; and unchanged for SENP3 and 7. UVA irradiation upregulates the mRNAs for all seven SENPs; In contrast to the mRNA levels for 7 SENPs, the expression levels for 6 SENPs (SENP1-3, 5-6 and 8) appeared down-regulated from 10% to 50%, and only SENP7 was slightly upregulated. CONCLUSION: Our results for the first time established the differentiation expression patterns of 7 de-sumoylation enzymes (SENPs) under treatment by GO or UVA, which provide preliminary data to link sumoylation to stress-induced cataractogenesis.