Edaravone improves the post-traumatic brain injury dysfunction in learning and memory by modulating Nrf2/are signal pathway

OBJECTIVES: To investigate the molecular mechanism of edaravone (EDA) in improving the post-traumatic brain injury (TBI) dysfunction in learning and memory. METHODS: In vitro and in vivo TBI models were established using hydrogen peroxide (H2O2) treatment for hippocampal nerve stem cells (NSCs) and surgery for rats, followed by EDA treatment. WST 1 measurement, methylthiazol tetrazolium assay, and flow cytometry were performed to determine the activity, proliferation, and apoptosis of NSCs, and malondialdehyde (MDA), lactic dehydrogenase (LDH), and reactive oxygen species (ROS) detection kits were used to analyze the oxides in NSCs. RESULTS: Following EDA pretreatment, NSCs presented with promising resistance to H2O2-induced oxidative stress, whereas NSCs manifested significant increases in activity and proliferation and a decrease in apoptosis. Meanwhile, for NSCs, EDA pretreatment reduced the levels of MDA, LDH, and ROS, with a significant upregulation of Nrf2/antioxidant response element (ARE) signaling pathway, whereas for EDA-treated TBI rats, a significant reduction was observed in the trauma area and injury to the hippocampus, with improvement in memory and learning performance and upregulation of Nrf2/ARE signaling pathway. CONCLUSIONS: EDA, by regulating the activity of Nrf2/ARE signal pathway, can improve the TBI-induced injury to NSCs and learning and memory dysfunction in rats.

Ação Quimiopreventiva dos Fitoquímicos por meio da Regulação do Fator de Transcrição Nrf2: Revisão Integrativa da Literatura: Revisão Integrativa de Literatura / Quimiopreventive Action of Phytochemicals through Regulation of Transcription Factor Nrf2: Integrative Literature Review / Acción Quimiopreventiva de los Fitoquímicos a través de la Regulación del Factor de Transcripción Nrf2: Revisión Integradora de la Literatura

Introdução: O fator nuclear eritroide 2 relacionado ao fator 2 (Nrf2) desempenha papel fundamental na expressão de genes mediados por elemento de resposta antioxidante (ERA); sendo assim, é uma via importante para proteger as células de substâncias carcinogênicas. Objetivo: Realizar uma revisão integrativa da literatura acerca da ação quimiopreventiva dos fitoquímicos por meio da regulação do fator de transcrição Nrf2. Método: O levantamento de artigos para a revisão integrativa da literatura sobre essa temática foi realizado nos periódicos indexados nas bases de dados: Google Acadêmico, PubMed, SciELO, ScienceDirect e SpringerLink, utilizando-se os descritores advindos do MeSH: fitoquímicos, radicais livres, estresse oxidativo, carcinogênese, quimioprevenção e Nrf2. Os critérios de seleção foram artigos publicados de 2000 a 2019, relacionados, ou que investiguem diretamente a atuação de fitoquímicos no fator de transcrição Nrf2, e a prevenção do desenvolvimento de câncer. Resultados: Foram selecionados 58 artigos que estavam relacionados com o objetivo da revisão. Os estudos revisados apontaram que fitoquímicos, tais como resveratrol, curcumina, isotiocianato, luteolina, entre outros, atuam na ativação da via Nrf2, utilizando diferentes mecanismos, sendo eles dependentes ou independentes da proteína repressora Kelch-Like Epichlorohydrin-Associated Protein 1. Conclusão: Diante disso, conclui-se que a modulação do fator de transcrição Nrf2 é um mecanismo que se configura como um importante mediador no que concerne compostos nocivos ao organismo humano, e que a atuação dos fitoquímicos nessa via contribui para a redução do risco de câncer. No entanto, ainda não são completamente elucidados todos os mecanismos utilizados pelos fitoquímicos, sendo necessários ulteriores estudos na área

Introduction: The nuclear factor erythroid 2-related factor 2 (NRF2) plays a fundamental role in the expression of genes mediated by antioxidant response element (ARE), thus it is an important pathway to protect the cells from carcinogenic substances. Objective: To perform an integrative literature review on the quimiopreventive action of phytochemicals through regulation of the transcription factor Nrf2. Method: Search of papers for the integrative literature review about this theme conducted in journals indexed in the databases: Academic Google, PubMed, SciELO, ScienceDirect and Springer Link, using the MeSH descriptors: phytochemicals, free radicals, oxidative stress, carcinogenesis, chemoprevention and Nrf2. The selection criteria were articles published from 2000 to 2019, related to or that directly investigate the role of phytochemicals in the transcription factor Nrf2, and the prevention of cancer development. Results: 58 articles were selected, all related to the objective of the review. The reviewed studies showed that phytochemicals, such as resveratrol, curcumin, isothiocyanate, luteolin, among others, act on the activation of the Nrf2 pathway, using different mechanisms, which are dependent or independent of the repressor protein Kelch-Like Epichlorohydrin-Associated Protein 1. Conclusion: Therefore, the conclusion is that the modulation of the transcription factor Nrf2 is a mechanism that configures itself as an important mediator for harmful compounds to the human organism, and that the action of phytochemicals, in this pathway, contributes to the reduction of cancer risk. However, all the mechanisms used by phytochemicals, are not completely elucidated, and further studies are needed in the area

Introducción: El factor nuclear eritroide 2 relacionado con el factor 2 (Nrf2) desenvuelve un papel fundamental en la expresión de los genes mediados por él elemento de respuesta antioxidante (ERA), por lo tanto, es una vía importante para proteger las células de las sustancias carcinógenas. Objetivo: Realizar una revisión integradora de la literatura sobre la acción quimiopreventiva de los fitoquímicos mediante la regulación del factor de transcripción Nrf2. Método: El levantamiento de artículos para la revisión integral de la literatura sobre este tema se realizó en revistas indexadas en las bases de datos: Google Académico, PubMed, Scielo, ScienceDirect y SpringerLink, usando los descriptores MeSH: fitoquímicos, radicales libres, estrés oxidativo, carcinogénesis, quimioprevención y Nrf2. Los criterios de selección fueron artículos publicados entre 2000 y 2019, relacionados o que investigan directamente el papel de los fitoquímicos en el factor de transcripción Nrf2 y la prevención del desarrollo del câncer. Resultados: 58 artículos relacionados con el objetivo de la revisión fueron seleccionados. Los estudios revisados mostraron que los fitoquímicos, como el resveratrol, la curcumina, el isotiocito, la luteolina, entre otros, actúan sobre la activación de la vía Nrf2, utilizando diferentes mecanismos, que son dependientes o independientes de la proteína represora Kelch-Like Epichlorohydrin-Associated Protein 1. Conclusión: Por lo tanto, se concluí que la modulación del factor de transcripción Nrf2 es un mecanismo que se configura como un importante mediador en relación con los compuestos nocivos para el cuerpo humano, y que la acción de los fitoquímicos en esta vía contribuye a reducir el riesgo de cáncer. Sin embargo, todos los mecanismos utilizados por los fitoquímicos aún no se han dilucidado por completo, por lo que se necesitan más estudios en esta área

Perfil lipídico y ensayos de las actividades antioxidante, insecticida y antialimentaria de la esponja marina Iotrochota birotulata (Iotrochotidae: Demospongiae) / Lipid profile and assays of the antioxidant, insecticide and antifeedant activities of the marine sponge Iotrochota birotulata (Iotrochotidae: Demospongiae)

Resumen Los mares ocupan aproximadamente tres cuartas partes de la tierra constituyendo el hábitat de una gran cantidad de especies de flora y fauna. Como parte de los organismos marinos, los phylum bioquímicamente más estudiados son Porífera, Cnidaria y más recientemente Ascomycota, de todos ellos se han aislado e identificado un gran número de compuestos químicos, los cuales han mostrado diferentes tipos de actividades biológicas, principalmente citotóxica, bactericida y antioxidante, entre otras. En esta investigación se estudió la esponja marina Iotrochota birotulata recolectada en Punta Bello-Córdoba en el Caribe colombiano lográndose determinar su perfil lipídico a partir del análisis de los correspondientes espectros de masas de sus compuestos constituyentes, fueron identificados 19 ácidos grasos, los cuales son reportados por primera vez para esta especie. Además fueron identificados 10 esteroles. Se encontraron ácidos grasos con cadenas carbonadas entre C15 y C27, mostrando una amplia diversidad estructural lo cual es característico del phylum porífera. En cuanto a los esteroles se encontraron estructuras principalmente con núcleos Δ5 y Δ7 que son los más abundantes en las esponjas. Se encontró moderada actividad antioxidante en la estabilización del catión radical ABTS+• y baja actividad en la estabilización del radical DPPH•, el mayor valor de inhibición se encontró en el extracto acuoso con un valor de IC50 de 43.9 µg/ml frente al catión radical ABTS+•, mostrando que I. birotulata puede producir compuestos con moderada actividad antioxidante. En el ensayo de actividad insecticida, se encontró afectación larval en los extractos metanólico y acuoso en todas las concentraciones y en todos los tiempos de exposición evaluados frente a Spodoptera frugiperda, también se determinó que estos extractos inducen un efecto antialimentario en las larvas como parte del mecanismo insecticida. Se determinaron las condiciones y los tiempos de exposición óptimos para la mejor eficiencia del ensayo, las cuales fueron de 3 000 µg/ml y 48 h para el extracto metanólico y 2 000 µg/ml y 24 h para el extracto acuoso, respectivamente. En conclusión, los extractos evaluados de I. birotulata mostraron moderada actividad antioxidante y buenas e interesantes actividades insecticida y antialimentaria.(AU)

Abstract It is estimated that three quarters of earth´s surface is occupied by the oceans. This constitutes the habitat of a large number of species of flora and fauna. As part of the marine organisms, the most studied biochemical phylum are Porifera, Cnidaria and more recently Ascomycota, from which a large number of chemical compounds have been isolated and identified. These have shown different types of biological activities, mainly cytotoxic, bactericidal and antioxidant. In this research the marine sponge Iotrochota birotulata collected in Punta Bello-Córdoba in the Colombian Caribbean was studied, and its lipid profile from the analysis of the corresponding mass spectra of its constituent compounds was determined, 19 fatty acids were identified, which are reported for the first time for this specie, besides 10 sterols were identified. Fatty acids with carbon chains between C15 and C27 were found, showing a wide structural diversity, which is characteristic of the porifera phylum. The sterols showed structures mainly with nucleus Δ5 and Δ7, which are the most abundant in sponges. Moderate antioxidant activity was found in the stabilization of the radical cation ABTS+• and low activity in the stabilization of the radical DPPH•, the highest inhibition value was found in the aqueous extract with an IC50 de 43.9 µg/ml against the radical cation ABTS+•, showing that I. birotulata can produce compounds with moderate antioxidant activity. In the insecticidal activity test, larval affectation was found in methanolic and aqueous extracts at all concentrations and at all times of exposure evaluated against Spodoptera frugiperda, it was also determined that these extracts induce an antifeedant effect on the larvae as part of the mechanism insecticide. The optimal conditions and exposure times were determined for the best efficiency of the assay, which were 3 000 µg/ml and 48 h for the methanolic extract and 2 000 µg/ml and 24 h for the aqueous extract, respectively. In conclusion, the evaluated extracts of the I. birotulata showed moderate antioxidant activity and good and interesting insecticidal and antifeedant activities.(AU)

Ajoene, a Major Organosulfide Found in Crushed Garlic, Induces NAD(P)H:quinone Oxidoreductase Expression Through Nuclear Factor E2-related Factor-2 Activation in Human Breast Epithelial Cells

BACKGROUND: NAD(P)H:quinone oxidoreductase-1 (NQO1) is a widely-distributed flavin adenine dinucleotide-dependent flavoprotein that promotes obligatory 2-electron reductions of quinones, quinoneimines, nitroaromatics, and azo dyes. This reduces quinone levels and thereby minimizes generation of excess reactive oxygen species (ROS) formed by redox cycling, and concurrent depletion of intracellular thiol pools. Ajoene is derived from crushed garlic. It is formed by a reaction involving two allicin molecules, and is composed of allyl sulfide and vinyl disulfide. Ajoene is present in two isomers, E- and Z-form. METHODS: Expression of antioxidant enzymes and nuclear factor E2-related factor-2 (Nrf2) was measured by Western blot analysis. NQO1 promoter activity was assessed by the luciferase reporter gene assay. ROS accumulation was monitored by using the fluorescence-generating probe 2′,7′-dichlorofluorescein diacetate. The intracellular glutathione levels were measured by using a commercially available kit. RESULTS: Z-ajoene significantly up-regulated the expression of representative antioxidant enzyme NQO1 in non-tumorigenic breast epithelial MCF-10A cells at non-toxic concentrations. Z-ajoene enhanced up-regulation and nuclear translocation of Nrf2, which plays a pivotal role in the induction of many genes encoding antioxidant enzymes and other cytoprotective proteins. Z-ajoene treatment also increased the activity of nqo1-promoter harboring antioxidant response element consensus sequences in MCF-10A cells. Silencing of Nrf2 by small interfering RNA abrogated ajoene-induced expression of NQO1. Z-ajoene activated extracellular signal-regulated kinase (ERK). Inhibition of ERK activation by U0126 abrogated ability of Z-ajoene to activate Nrf2 and to induce NQO1 expression. Intracellular ROS accumulation was observed after treatment with Z-ajoene, whereas the E-isoform was not effective. The inhibition of ROS by treatment with N-acetylcysteine, a radical scavenger, abrogated Z-ajoene-induced expression of NQO1 as well as activation of ERK and Nrf2, suggesting that Z-ajoene augments the Nrf2-dependent antioxidant defense via ROS generation and ERK activation. CONCLUSIONS: Z-ajoene induces NQO1 expression in MCF-10A cells through ROS-mediated activation of Nrf2.

4′-O-β-D-Glucosyl-5-O-Methylvisamminol Attenuates Pro-Inflammatory Responses and Protects against Oxidative Damages

We attempted to examine anti-inflammatory and anti-oxidant effects of 4′-O-β-D-glucosyl-5-O-methylvisamminol (GOMV), the first epigenetic inhibitor of histone phosphorylation at Ser10. While GOMV did not affect the viability of murine macrophage RAW 264.7 cells, it significantly suppressed lipopolysaccharide (LPS)-induced generation of prostaglandin E₂ (PGE₂) and nitric oxide (NO) through transcriptional inhibition of cyclooxygenase-2 (COX-2) and inducible nitric oxide synthase (iNOS). GOMV also scavenged free radicals in vitro, increased NF-E2-related factor 2 (NRF2), and activated antioxidant response element (ARE), thereby resulting in the induction of phase II cytoprotective enzymes in human keratinocyte HaCaT cells. Finally, GOMV significantly protected HaCaT cells against 12-O-tetradecanoylphorbol-13-acetate (TPA)-induced oxidative intracellular damages. Together, our results illustrate that GOMV possesses anti-inflammatory and anti-oxidant activity.

Assessment of homeopathic medicines on survival and antioxidant response in white shrimp Litopenaeus vannamei / Evaluación de medicamentos homeopáticos en la supervivencia y respuesta antioxidante del camarón blanco Litopenaeus vannamei infectado con Vibrio parahaemolyticus

ABSTRACT Objective. Evaluating the effect of homeopathic medicines on survival and activity of the superoxide dismutase (SOD) enzyme in white shrimp Litopenaeus vannamei subjected to infection with the pathogenic Vibrio parahaemolyticus. Materials and methods . The average lethal dosage (LD50) was determined for the pathogen strain in L. vannamei juveniles under immersion (Imm), injection (Inj) and incision + immersion (Inc+Imm) methods. Four treatments were applied: (1) Mix CIB®-HOM Heel-Mix (TH1), constituted by equal v/v ratio, of Cyme-Heel®, Gal-Heel®, Hepa-Heel®, Mucs-Heel® and Chol-Heel®; (2) Mix CIB®-HOM Pav-Mix (TH2), constituted by the same v/v ratio of Passiflora incarnata, Valeriana officinalis, Zincum valerianicum and Ignatia amara (Similia®); (3) Heel-Mix/Pav-Mix (TH3) consisting of a 1:1 v/v combination of the TH1 and TH2 treatments, and (4) ViT-Mix (TH4), constituted by Vidatox®, and a control (not treated/infected). Results. While applying the methods Imm, Inj and Inc+Imm, LD50 was 0.9 x 106; 0.6 x 106 and 0.5 x 106 UFC.mL-1, respectively. At the end of the challenge, the groups treated with TH2, TH3 and TH4 had a greater survival rate to that of the control group (p<0.05). Moreover, these two last treatments showed a greater SOD activity with respect to the control group (p<0.05). Conclusions. The homeopathic treatments (TH3 and TH4) increased survival and SOD activity in L. vannamei juveniles challenged with V. parahaemolyticus, which suggests that the homeopathic treatments employed had the potential as an alternative for the control of V. parahaemolyticus and its associated diseases, including the early mortality syndrome in shrimp farming.

RESUMEN Objetivo. Evaluar el efecto de medicamentos homeopáticos sobre la supervivencia y actividad de la enzima superóxido dismutasa (SOD) del camarón Litopenaeus vannamei sometido a infección con Vibrio parahaemolyticus. Materiales y métodos. Se determinó la dosis letal media (DL50) para la cepa patógena en juveniles de L. vannamei, bajo los método de inmersión (Inm), inyección (Iny) e incisión + inmersión (Inc+Inm). Luego el efecto de cuatro medicamentos homeopáticos sobre juveniles de L. vannamei retados con Vibrio parahaemolyticus fue evaluado usando el índice la supervivencia y la actividad SOD. Se aplicaron cuatro tratamientos: (1) Mezcla CIB®-HOM Heel-Mix (TH1), constituido por igual proporción v/v, de Cyme-Heel®, Gal-Heel®, Hepa-Heel®, Mucs-Heel® y Chol-Heel®; (2) Mezcla CIB®-HOM Pav-Mix (TH2), constituido por igual proporción v/v de Passiflora incarnata, Valeriana officinalis, Zincum valerianicum e Ignatia amara (Similia®); (3) Heel-Mix/Pav-Mix (TH3) constituido por una combinación 1:1 v/v de los tratamientos TH1 y TH2, y (4) ViT-Mix (TH4), constituido por Vidatox®, y un control (no tratado/infectado). Resultados. Al aplicar los método Inm, Iny e Inc+Inm la DL50 fue de 0.9 x 106; 0.6 x 106 y 0.5 x 106 UFC.mL-1, respectivamente. Los camarones tratados con TH3 y TH4 presentaron una mayor actividad de SOD con respecto al grupo control (p<0.05). Al final del reto, los grupos TH2, TH3 y TH4 tuvieron una supervivencia mayor a la del grupo control (p<0.05). Conclusiones. Los tratamientos homeopáticos (TH3 y TH4), aumentaron la actividad de la enzima SOD y la supervivencia en juveniles de L. vannamei, retados con V. parahaemolyticus. Esto sugiere que los tratamientos homeopáticos empleados tienen potencial como alternativa para el control de V. parahaemolyticus y sus enfermedades asociadas, incluido el síndrome de mortalidad temprana en el cultivo del camarón.

Inhibitory Effect of Ginsenosides Rh1 and Rg2 on Oxidative Stress in LPS-Stimulated RAW 264.7 Cells

Minor ginsenosides Rh1 and Rg2 were isolated from Korean red ginseng and reported to have various biological effects on anti-inflammatory and anti-stress activities. However, the effects of Rh1 and Rg2 on antioxidant activity and their regulatory effects on the antioxidant enzymes have not been studied. Since oxidative stress is one of the major toxic inflammatory responses stimulated by lipopolysaccharides (LPS), the present study investigated the role of minor ginsenosides Rh1 and Rg2 on antioxidant effects in LPS-treated RAW 264.7 cells. In this study, we found that treatment with ginsenosides Rh1 and Rg2 strongly inhibited LPS-stimulated intracellular ROS production in cells. Luciferase assay showed that treatment with LPS reduced antioxidant response element (ARE) encoding the pARE-luc promoter activity, while ginsenosides inhibited the pARE-luc promoter activity. Moreover, ginsenosides Rh1 and Rg2 exhibited anti-oxidative activity in LPS-induced cells by upregulating antioxidant enzymes including superoxide dismutase, catalase, and glutathione peroxidase. Our results suggest that minor ginsenosides Rh1 and Rg2 may be potential bio-active compounds for antioxidative effects by inhibiting the generation of ROS in RAW 264.7 cells.

Synthesis and cytotoxicity evaluation of 3-amino-2-hydroxypropoxygenistein derivatives / 中国天然药物

Soy isoflavones exhibit various biological activities, such as antioxidant, anti-tumor, anti-inflammatory, and cardiovascular protective effects. The present study was designed to investigate the effects of sixteen synthesized 3-amino-2-hydroxypropoxy genistein derivatives on cell proliferation and activation of Nrf2 (Nuclear factor erythroid 2-related factor 2)/ARE (antioxidant response elements) pathway in human cancer cell lines. Most of the tested compounds exerted greater cytotoxic activity than genistein, as measured by MTT assay. Moreover, compound 8c showed the highest ARE-luciferase reporter activity among the test compounds. It strongly promoted Nrf2 nuclear translocation and up-regulated the expression of total Nrf2 and downstream targets NQO-1 and HO-1 at protein level. The present study may provide a basis for the application of isoflavone derivatives as Nrf2/ARE pathway inducers for cancer therapy and cancer prevention.

Expression and significance of Nrf2/ARE pathway ralated factors in the HepG2 cell model of steatosis / 中国应用生理学杂志

<p><b>OBJECTIVE</b>To explore a new method of establishing HepG2 cell model of steatosis and observe the expression and significance of nuclear factor erythroid-2p45-related factor 2(Nrf2)/antioxidative response element (ARE) pathway related factors in HepG2 cells of steatosis.</p><p><b>METHODS</b>HepG2 cells were induced with DMEM containing 25% fetal bovine serum, 0.1% MCT/LCT Fat Emulsion and 0.1 mmol/L free fatty acid (FFA) at different stages and the control group cells were cultured with normal DMEM medium. After the cell models were successfully established, lipid droplets in cytoplasm were observed with Oil Red 0 staining, and the triglyceride (TG) accumulation in HepG2 cells were tested by biochemical assay. Intracellular reactive oxygen species (ROS) concentration were detected by flow cytometry. Nitric oxide (NO), superoxide dismutase(SOD), malonyldialdehyde(MDA) and glutathione peroxidase(GSH-Px) were tested by biological reagent kit, while the protein expression of nuclear factor erythroid-2p45-related factor 2(Nrf2), heme oxygenase-1 (HO-1) and</p><p><b>NAD(P)H</b>quinone oxidoreductase-1(NQO1) were analyzed by Western blot.</p><p><b>RESULTS</b>Compared with that in the control group, red cytoplasmic lipid droplets were visible in model group; TG,ROS, NO, MDA concentration (P < 0.05, P < 0.01) and the protein expression of Nrf2, HO-1 and NQO1 (P < 0.05, P < 0.01)were significantly higher in model group, while SOD, GSH-Px concentration reduced significantly (P < 0.01).</p><p><b>CONCLUSION</b>The in vitro cell model of steatosis and oxidative stress was successfully established. The activation of Nrf2/ARE pathway related factors maybe relevant to the overreaction of oxidative stress in HepG2 cells of steatosis.</p>

Protopanaxatriol Ginsenoside Rh1 Upregulates Phase II Antioxidant Enzyme Gene Expression in Rat Primary Astrocytes: Involvement of MAP Kinases and Nrf2/ARE Signaling

Oxidative stress activates several intracellular signaling cascades that may have deleterious effects on neuronal cell survival. Thus, controlling oxidative stress has been suggested as an important strategy for prevention and/or treatment of neurodegenerative diseases. In this study, we found that ginsenoside Rh1 inhibited hydrogen peroxide-induced reactive oxygen species generation and subsequent cell death in rat primary astrocytes. Rh1 increased the expression of phase II antioxidant enzymes, such as heme oxygenase-1 (HO-1), NAD(P)H:quinone oxidoreductase 1, superoxide dismutase-2, and catalase, that are under the control of Nrf2/ARE signaling pathways. Further mechanistic studies showed that Rh1 increased the nuclear translocation and DNA binding of Nrf2 and c-Jun to the antioxidant response element (ARE), and increased the ARE-mediated transcription activities in rat primary astrocytes. Analysis of signaling pathways revealed that MAP kinases are important in HO-1 expression, and act by modulating ARE-mediated transcriptional activity. Therefore, the upregulation of antioxidant enzymes by Rh1 may provide preventive therapeutic potential for various neurodegenerative diseases that are associated with oxidative stress.

Ethanol Extract of Cirsium japonicum var. ussuriense Kitamura Exhibits the Activation of Nuclear Factor Erythroid 2-Related Factor 2-dependent Antioxidant Response Element and Protects Human Keratinocyte HaCaT Cells Against Oxidative DNA Damage

Keratinocytes are constantly exposed to extracellular insults, such as ultraviolet B, toxic chemicals and mechanical stress, all of which can facilitate the aging of keratinocytes via the generation of intracellular reactive oxygen species (ROS). Nuclear factor erythroid 2-related factor 2 (Nrf2) is a transcription factor that plays a critical role in protecting keratinocytes against oxidants and xenobiotics by binding to the antioxidant response element (ARE), a cis-acting element existing in the promoter of most phase II cytoprotective genes. In the present study, we have attempted to find novel ethanol extract(s) of indigenous plants of Jeju island, Korea that can activate the Nrf2/ARE-dependent gene expression in human keratinocyte HaCaT cells. As a result, we identified that ethanol extract of Cirsium japonicum var. ussuriense Kitamura (ECJUK) elicited strong stimulatory effect on the ARE-dependent gene expression. Supporting this observation, we found that ECJUK induced the expression of Nrf2, hemoxygenase-1, and NAD(P)H:quinone oxidoreductase-1 and this event was correlated with Akt1 phosphorylation. We also found that ECJUK increased the intracellular reduced glutathione level and suppressed 12-O-tetradecanoylphorbol acetate-induced 8-hydroxyguanosine formation without affecting the overall viability. Collectively, our results provide evidence that ECJUK can protect against oxidative stress-mediated damages through the activation of Nrf2/ARE-dependent phase II cytoprotective gene expression.

Involvement of Heme Oxygenase-1 in Orexin-A-induced Angiogenesis in Vascular Endothelial Cells

The cytoprotective enzyme heme oxygenase-1 (HO-1) influences endothelial cell survival, proliferation, inflammatory response, and angiogenesis in response to various angiogenic stimuli. In this study, we investigate the involvement of HO-1 in the angiogenic activity of orexin-A. We showed that orexin-A stimulates expression and activity of HO-1 in human umbilical vein endothelial cells (HUVECs). Furthermore, we showed that inhibition of HO-1 by tin (Sn) protoporphryin-IX (SnPP) reduced orexin-A-induced angiogenesis in vivo and ex vivo. Orexin-A-stimulated endothelial tube formation and chemotactic activity were also blocked in SnPP-treated vascular endothelial cells. Orexin-A treatment increased the expression of nuclear factor erythroid-derived 2 related factor 2 (Nrf2), and antioxidant response element (ARE) luciferase activity, leading to induction of HO-1. Collectively, these findings indicate that HO-1 plays a role as an important mediator of orexin-A-induced angiogenesis, and provide new possibilities for therapeutic approaches in pathophysiological conditions associated with angiogenesis.

Effects of acute and chronic administration of methylprednisolone on oxidative stress in rat lungs / Efeitos da administração aguda e crônica de metilprednisolona no estresse oxidativo em pulmões de ratos

Objective: To determine the effects of acute and chronic administration of methylprednisolone on oxidative stress, as quantified by measuring lipid peroxidation (LPO) and total reactive antioxidant potential (TRAP), in rat lungs. Methods: Forty Wistar rats were divided into four groups: acute treatment, comprising rats receiving a single injection of methylprednisolone (50 mg/kg i.p.); acute control, comprising rats i.p. injected with saline; chronic treatment, comprising rats receiving methylprednisolone in drinking water (6 mg/kg per day for 30 days); and chronic control, comprising rats receiving normal drinking water. Results: The levels of TRAP were significantly higher in the acute treatment group rats than in the acute control rats, suggesting an improvement in the pulmonary defenses of the former. The levels of lung LPO were significantly higher in the chronic treatment group rats than in the chronic control rats, indicating oxidative damage in the lung tissue of the former. Conclusions: Our results suggest that the acute use of corticosteroids is beneficial to lung tissue, whereas their chronic use is not. The chronic use of methylprednisolone appears to increase lung LPO levels. .

Objetivo: Determinar os efeitos da administração aguda e crônica de metilprednisolona no estresse oxidativo, por meio da quantificação da peroxidação lipídica (POL) e do potencial antioxidante reativo total (PART), em pulmões de ratos. Métodos: Quarenta ratos Wistar foram divididos em quatro grupos: tratamento agudo, com ratos recebendo uma dose única de metilprednisolona (50 mg/kg i.p.); controle agudo, com ratos recebendo injeção unida de salina; tratamento crônico, com ratos recebendo metilprednisolona v.o. na água do bebedouro (6 mg/kg por dia durante 30 dias; e controle crônico, com ratos recebendo água de bebedouro normal). Resultados: Os níveis de PART foram significativamente maiores no grupo tratamento agudo que no grupo controle agudo, sugerindo uma melhora do sistema de defesa pulmonar. Os níveis de POL foram significativamente maiores no grupo tratamento crônico que no grupo controle crônico, indicando dano oxidativo no tecido pulmonar. Conclusões: Nossos resultados sugerem que o uso agudo de corticoides foi benéfico aos tecidos pulmonares, enquanto seu uso crônico não o foi. O uso crônico de metilprednisolona parece aumentar os níveis pulmonares da POL. .

Impacto da ingestão dietética e do estresse oxidativo em pacientes com doença trofoblástica gestacional / Impact of dietary intake and oxidative stress in patients with gestational trophoblastic disease

A doença trofoblástica gestacional (DTG) é um termo aplicado a um grupo de tumores relacionados à gestação, caracterizando formas benignas (mola hidatiforme - MH) e malignas (neoplasia trofoblástica gestacional - NTG). O estresse oxidativo, a ingestão dietética e o estado nutricional da gestante constituem possíveis fatores de risco para a ocorrência da MH e sua progressão para NTG. A presente revisão tem como objetivo abordar a relação entre essas entidades. Apesar de haver evidências mostrando que a ingestão dietética e o estresse oxidativo possam ter papéis relevantes na etiopatogênese da DTG, ainda são necessários muitos estudos para uma melhor investigação da atuação desses agentes na gravidez molar e suas sequelas malignas e proliferativas.(AU)

The gestational trophoblastic disease (GTD) is a term applied to a rare group of pregnancy related tumors of benign forms (hydatidiform mole - HM) as well as malignant ones (gestational trophoblastic neoplasia - GTN). Oxidative stress, dietetic intake and nutritional status of pregnant women are possible risk factors for the occurrence of HM and its progression to GTN. This review aims to address the relationship between these entities. Besides the evidences supporting that dietetic intake and oxidative stress may have important roles in the GTD etiopathogenesis, many studies are needed to better investigate the role of these agents in molar pregnancy and their proliferative and malignant sequelae.(AU)

Arctigenin Increases Hemeoxygenase-1 Gene Expression by Modulating PI3K/AKT Signaling Pathway in Rat Primary Astrocytes

In the present study, we found that the natural compound arctigenin inhibited hydrogen peroxide-induced reactive oxygen species (ROS) production in rat primary astrocytes. Since hemeoxygenase-1 (HO-1) plays a critical role as an antioxidant defense factor in the brain, we examined the effect of arctigenin on HO-1 expression in rat primary astrocytes. We found that arctigenin increased HO-1 mRNA and protein levels. Arctigenin also increases the nuclear translocation and DNA binding of Nrf2/c-Jun to the antioxidant response element (ARE) on HO-1 promoter. In addition, arctigenin increased ARE-mediated transcriptional activities in rat primary astrocytes. Further mechanistic studies revealed that arctigenin increased the phosphorylation of AKT, a downstream substrate of phosphatidylinositol 3-kinase (PI3K). Treatment of cells with a PI3K-specific inhibitor, LY294002, suppressed the HO-1 expression, Nrf2 DNA binding and ARE-mediated transcriptional activities in arctigenin-treated astrocyte cells. The results collectively suggest that PI3K/AKT signaling pathway is at least partly involved in HO-1 expression by arctigenin via modulation of Nrf2/ARE axis in rat primary astrocytes.

Tanshinone IIA protects against triptolide-induced liver injury via Nrf2/ARE activation / 药学学报

The aim of this study is to investigate the protection effect of tanshinone IIA (Tan) against triptolide (TP)-induced liver injury and the mechanisms involved. Acute liver injury was induced by intraperitoneal injection of TP (1 mg x kg(-1)) in mice. The activities of AST, ALT and LDH in serum and the levels of GSH, GST, GSH-PX, SOD, CAT and MDA in liver tissue were detected. The histopathological changes of liver tissues were observed after HE staining. Nrf2 translocation in liver tissue was detected by Western blotting, and real-time PCR was used to measure the expression levels of GCLC, NQO1 and HO-1 mRNA. The results showed that pretreatment with Tan significantly prevented the TP induced liver injury as indicated by reducing the activities of AST, ALT and LDH (P < 0.01). Tan pretreatment also prevented TP-induced oxidative stress in the mice liver by inhibiting MDA and restoring the levels of GSH, GST, SOD and CAT (P < 0.05). Parallel to these changes, pretreatment with Tan could attenuate histopathologic changes induced by TP. Furthermore, the results indicated that Tan pretreatment caused nuclear accumulation of Nrf2 as well as induction of mRNA expression of antioxidant response element (ARE)-driven genes such as GCLC, NQO1 and HO-1. These results indicated that Tan could protect against TP-induced acute liver injury via the activation of Nrf2/ARE pathway.

Expression and antioxidation of Nrf2/ARE pathway in traumatic brain injury

OBJECTIVE@#To explore the expression of Nrf2/ARE pathway in hindbrain tissue after the traumatic brain injury (TBI) and its anti-oxidative stress effect in the secondary nerve injury.@*METHODS@#The mice with Nrf2 gene knockout were used for the establishment of brain injury model. The experimental animals were divided into four groups: (Nrf2(+)/(+)) sham-operation group, (Nrf2(+)/(+)) brain injury group, (Nrf2(-)/(-)) sham-operation group and (Nrf2(-)/(-)) brain injury group. The specimen 24 h after cerebral trauma was selected. Then RT-PCR method was adopted to detect the expression of Nrf2 mRNA in brain; Western blotting method was adopted to detect the levels of Nrf2, HO-1 and NQO1 proteins in brain; ELISA method was adopted to detect the oxidative stress indicators: protein carbonyls, 4-hydroxy-2-nonenal (4-HNE) and 8-hydroxy-2'-deoxyguanosine (8-OHdG).@*RESULTS@#The Nrf2 mRNA and protein of Nrf2(-)/(-) mice were not expressed, and the difference of the relative amount of Nrf2 mRNA between Nrf2(+)/(+) TBI group and Nrf2(+)/(+) sham-operation group was not statistically significant (P>0.05); the level of Nrf2 protein in Nrf2(+)/(+) TBI group increased significantly compared with the Nrf2(+)/(+) sham-operation group (P0.05); there was only a little amount of expression of protein carbonyls, 4-HNE and 8-OHdG proteins in brain tissues in the Nrf2(+)/(+) and Nrf2(-)/(-) sham-operation groups, and the difference was not statistically significant (P>0.05); after brain injury, the three oxidative stress indicators were significantly up-regulated in the Nrf2(+)/(+) and Nrf2(-)/(-) groups, and the up-regulation of the latter group was more significant (P<0.01).@*CONCLUSIONS@#After TBI the Nrf2/ARE pathway is activated and the activity of Nrf2 transcription regulation increases. However, the regulation dose not occur in the gene transcription level and only could increase the Nrf2 protein level, while the mRNA expression level has no obvious change. The nerve cell protective effect of Nrf2/ARE pathway in TBI achieves through inhibiting the oxidative stress injuries.

Protection by Chrysanthemum zawadskii extract from liver damage of mice caused by carbon tetrachloride is maybe mediated by modulation of QR activity

Our previous study demonstrated that methanolic extract of Chrysanthemum zawadskii Herbich var. latilobum Kitamura (Compositae) has the potential to induce detoxifying enzymes such as NAD(P)H:(quinone acceptor) oxidoreductase 1 (EC 1.6.99.2) (NQO1, QR) and glutathione S-transferase (GST). In this study we further fractionated methanolic extract of Chrysanthemum zawadskii and investigated the detoxifying enzyme-inducing potential of each fraction. The fraction (CZ-6) shown the highest QR-inducing activity was found to contain (+)-(3S,4S,5R,8S)-(E)-8-acetoxy-4-hydroxy-3-isovaleroyloxy-2-(hexa-2,4-diynyliden)-1,6-dioxaspiro [4,5] decane and increased QR enzyme activity in a dose-dependent manner. Furthermore, CZ-6 fraction caused a dose-dependent enhancement of luciferase activity in HepG2-C8 cells generated by stably transfecting antioxidant response element-luciferase gene construct, suggesting that it induces antioxidant/detoxifying enzymes through antioxidant response element (ARE)-mediated transcriptional activation of the relevant genes. Although CZ-6 fraction failed to induce hepatic QR in mice over the control, it restored QR activity suppressed by CCl4 treatment to the control level. Hepatic injury induced by CCl4 was also slightly protected by pretreatment with CZ-6. In conclusion, although CZ-6 fractionated from methanolic extract of Chrysanthemum zawadskii did not cause a significant QR induction in mice organs such as liver, kidney, and stomach, it showed protective effect from liver damage caused by CCl4.

Antioxidant capacity in relationship to serum lipid peroxides levels in healthy elderly of Mexico City / Capacidad antioxidante en relación a niveles séricos de lipoperóxidos en ancianos sanos de la ciudad de México

The aging is one of the factors that cause decrease in the antioxidant capacity. Likewise, it has been proposed that subjects exposed permanently to air pollution develop deficient antioxidant capacity to oxidative stress (OxS). This study aimed to analyze the antioxidant capacity against elevated lipid peroxides in healthy elderly of Mexico City. 105 adults (44 ± 10.8 years) and 126 elderly subjects were studied (68 ± 7.1 years); residents of Mexico City (clinically healthy, non-smokers, non-vitamin supplement takers) who had lived in the city for >10 years. Plasma lipoperoxides (LPO), total antioxidant status (TAS), the activity of red blood cells superoxide dismutase (SOD), and plasma glutathione peroxidase (GPx), were studied in all subjects. LPO levels were found significantly higher (p < 0.05) in the elderly subjects in comparison with the adults; in addition, TAS and GPx were higher in adults than among the elderly people (p < 0.0001). Nevertheless, SOD was similar in both groups (p = 0.346). These findings reveal that the elderly residents of Mexico City have TAS and GPx lower than adults, and similar SD activity, probably due to the fact that these antioxidants are neutralizing the higher LPO levels of elderly people. Therefore, this mechanism could be considered as an efficient antioxidant capacity in the elderly, as response to high LPO levels, since the health status, mortality prevalence and life span life of the older people of Mexico City are similar or better than other cities of Mexican Republic.

Se ha propuesto que los sujetos expuestos permanentemente a la contaminación ambiental tienen una deficiente capacidad para contrarrestar el estrés oxidativo (EOx) y que el envejecimiento es un factor causante de dicha alteración. El objetivo de este estudio fue analizar la capacidad antioxidante contra el aumento de lipoperóxidos (LPO) en adultos mayores sanos de la ciudad de México. Se estudiaron 105 adultos residentes de la ciudad de México (44 ± 10,8 años) y 126 adultos mayores (68 ± 7,1 años) clínicamente sanos, no fumadores, sin ingesta de vitaminas antioxidantes, con residencia en esta ciudad por más 10 años. Se cuantificó a todos los sujetos los LPO plasmáticos, capacidad sérica antioxidante total (AT), actividad eritrocitaria de superóxido dismutasa (SOD) y plasmática de glutatión peroxidasa (GPx). Se encontró que los niveles de LPO fueron más altos en los adultos mayores comparados con los jóvenes (p < 0,05); asimismo, AT y GPx fueron mayores en los jóvenes (p < 0,0001). La SOD fue similar en ambos grupos (p = 0,346). Estos hallazgos revelan que los ancianos residentes de la ciudad de México tienen concentraciones más bajas de AT y GPx en comparación con los adultos y una actividad similar de la SOD, debido probablemente a que estos antioxidantes están neutralizando los niveles más altos de los LPO de los ancianos. Por lo tanto, este mecanismo podría ser considerado como una capacidad antioxidante eficiente en los ancianos como respuesta a los altos niveles de LPO, ya que el estado de salud, prevalencia de mortalidad y longevidad de los adultos mayores de la ciudad de México es similar o mejor al de los residentes de otros estados de la República Mexicana.