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1.
Molecules ; 26(12)2021 Jun 12.
Artigo em Inglês | MEDLINE | ID: mdl-34204601

RESUMO

Interferonopathies are rare genetic conditions defined by systemic inflammatory episodes caused by innate immune system activation in the absence of pathogens. Currently, no targeted drugs are authorized for clinical use in these diseases. In this work, we studied the contribution of sulforaphane (SFN), a cruciferous-derived bioactive molecule, in the modulation of interferon-driven inflammation in an immortalized human hepatocytes (IHH) line and in two healthy volunteers, focusing on STING, a key-component player in interferon pathway, interferon signature modulation, and GSTM1 expression and genotype, which contributes to SFN metabolism and excretion. In vitro, SFN exposure reduced STING expression as well as interferon signature in the presence of the pro-inflammatory stimulus cGAMP (cGAMP 3 h vs. SFN+cGAMP 3 h p value < 0.0001; cGAMP 6 h vs. SFN+cGAMP 6 h p < 0.001, one way ANOVA), restoring STING expression to the level of unstimulated cells. In preliminary experiments on healthy volunteers, no appreciable variations in interferon signature were identified after SFN assumption, while only in one of them, presenting the GSTM1 wild type genotype related to reduced SFN excretion, could a downregulation of STING be recorded. This study confirmed that SFN inhibits STING-mediated inflammation and interferon-stimulated genes expression in vitro. However, only a trend towards the downregulation of STING could be reproduced in vivo. Results obtained have to be confirmed in a larger group of healthy individuals and in patients with type I interferonopathies to define if the assumption of SFN could be useful as supportive therapy.


Assuntos
Inflamação/metabolismo , Isotiocianatos/farmacologia , Sulfóxidos/farmacologia , Adulto , Linhagem Celular Tumoral , Feminino , Expressão Gênica/efeitos dos fármacos , Genótipo , Glutationa Transferase/metabolismo , Voluntários Saudáveis , Hepatócitos/efeitos dos fármacos , Hepatócitos/metabolismo , Humanos , Imunidade Inata/efeitos dos fármacos , Inflamação/tratamento farmacológico , Interferons/efeitos adversos , Interferons/genética , Interferons/farmacologia , Isotiocianatos/metabolismo , Masculino , Proteínas de Membrana/metabolismo , Sulfóxidos/metabolismo
2.
Ecotoxicol Environ Saf ; 221: 112469, 2021 Sep 15.
Artigo em Inglês | MEDLINE | ID: mdl-34198190

RESUMO

Glutathione S-transferase (GST) is the key enzyme in glutathione (GSH) synthesis, and plays a crucial role in copper (Cu) detoxification. Nonetheless, its regulatory mechanisms remain largely unclear. In this study, we identified a Cu-induced glutathione S-transferase 1 (TaGST1) gene in wheat. Yeast one-hybrid (Y1H) screened out TaWRKY74, which was one member from the WRKY transcription factor family. The bindings between TaGST1 promoter and TaWRKY74 were further verified by using another Y1H and luciferase assays. Expression of TaWRKY74 was induced more than 30-folds by Cu stress. Functions of TaWRKY74 were tested by using transiently silence methods. In transiently TaWRKY74-silenced wheat plants, TaWRKY74 and TaGST1 expression, GST activity, and GSH content was significantly inhibited by 25.68%, 19.88%, 27.66%, and 12.68% in shoots, and 53.81%, 52.11%, 23.47%, and 17.11% in roots, respectively. However, contents of hydrogen peroxide, malondialdehyde, or Cu were significantly increased by 2.58%, 12.45%, or 37.74% in shoots, and 25.24%, 53.84%, and 103.99% in roots, respectively. Notably, exogenous application of GSH reversed the adverse effects of transiently TaWRKY74-silenced wheat plants during Cu stress. Taken together, our results suggesting that TaWRKY74 regulated TaGST1 expression and affected GSH accumulation under Cu stress, and could be useful to ameliorate Cu toxicity for crop food safety.


Assuntos
Cobre/toxicidade , Glutationa Transferase/metabolismo , Glutationa/metabolismo , Proteínas de Plantas/metabolismo , Fatores de Transcrição/metabolismo , Triticum/efeitos dos fármacos , Fatores de Transcrição/genética , Triticum/genética , Triticum/metabolismo , Técnicas do Sistema de Duplo-Híbrido , Leveduras/genética
3.
BMC Plant Biol ; 21(1): 287, 2021 Jun 24.
Artigo em Inglês | MEDLINE | ID: mdl-34167489

RESUMO

BACKGROUND: Phosphoenolpyruvate carboxylase (PEPC) plays an important role in the primary metabolism of higher plants. Several studies have revealed the critical importance of PEPC in the interaction of carbon and nitrogen metabolism. However, the function mechanism of PEPC in nitrogen metabolism is unclear and needs further investigation. RESULTS: This study indicates that transgenic rice expressing the sugarcane C4-PEPC gene displayed shorter primary roots and fewer crown roots at the seedling stage. However, total nitrogen content was significantly higher in transgenic rice than in wild type (WT) plants. Proteomic analysis revealed that there were more differentially expressed proteins (DEPs) responding to nitrogen changes in transgenic rice. In particular, the most enriched pathway "glutathione (GSH) metabolism", which mainly contains GSH S-transferase (GST), was identified in transgenic rice. The expression of endogenous PEPC, GST and several genes involved in the TCA cycle, glycolysis and nitrogen assimilation changed in transgenic rice. Correspondingly, the activity of enzymes including GST, citrate synthase, 6-phosphofructokinase, pyruvate kinase and ferredoxin-dependent glutamate synthase significantly changed. In addition, the levels of organic acids in the TCA cycle and carbohydrates including sucrose, starch and soluble sugar altered in transgenic rice under different nitrogen source concentrations. GSH that the substrate of GST and its components including glutamic acid, cysteine and glycine accumulated in transgenic rice. Moreover, the levels of phytohormones including indoleacetic acid (IAA), zeatin (ZT) and isopentenyladenosine (2ip) were lower in the roots of transgenic rice under total nutrients. Taken together, the phenotype, physiological and biochemical characteristics of transgenic rice expressing C4-PEPC were different from WT under different nitrogen levels. CONCLUSIONS: Our results revealed the possibility that PEPC affects nitrogen metabolism through regulating GST, which provide a new direction and concepts for the further study of the PEPC functional mechanism in nitrogen metabolism.


Assuntos
Glutationa Transferase/metabolismo , Nitrogênio/metabolismo , Oryza/enzimologia , Fosfoenolpiruvato Carboxilase/metabolismo , Saccharum/enzimologia , Carbono/metabolismo , Oryza/genética , Oryza/metabolismo , Fosfoenolpiruvato Carboxilase/genética , Plantas Geneticamente Modificadas , Proteômica , Saccharum/genética , Transcriptoma
4.
Sci Total Environ ; 778: 146375, 2021 Jul 15.
Artigo em Inglês | MEDLINE | ID: mdl-34030372

RESUMO

Bronopol and Detarox® AP are broad spectrum antimicrobial biocides of growing interest for the aquaculture sector. While their effectiveness against aquatic pathogens has been demonstrated, toxicity data on wild or farmed species are still lacking, as is information on their potential environmental risk for aquatic ecosystems. With this study, we assessed the acute and sublethal toxicity of Bronopol and Detarox® AP in the freshwater bivalve Sinanodonta woodiana and their theoretical risk for aquatic ecosystem. The 96-h median lethal concentration (LC50) was determined using the acute toxicity test, while for the sublethal toxicity test the bivalves were exposed to two concentrations for 14 days of Bronopol (2.5 and 50 mg/L) and Detarox® AP (1.11 and 22.26 mg/L) followed by a 14-day withdrawal period. Biocide-mediated oxidative processes were investigated via a panel of oxidative stress biomarkers (malondialdehyde, superoxide dismutase, catalase, glutathione peroxidase, glutathione S-transferase). Theoretical environmental risk assessment of both biocides, with predicted concentration of no effect (PNEC), expected theoretical concentration (TEC) in the environment, and risk quotient (RQ) was performed. TEC was calculated using a model based on the size of the aquaculture facility and the receiving basin, the estimated quantity of biocide dissolved in water, and published data on biocide stability in water. Although the LC50 was higher for Bronopol (2440 mg/L) than for Detarox® AP (126 mg/L), fluctuations in oxidative stress biomarkers levels indicated that both biocides exert a slight oxidative pressure on S. woodiana. Theoretical environmental risk assessment suggested a muted risk with Detarox® AP and greater eco-sustainability compared to Bronopol.


Assuntos
Desinfetantes , Poluentes Químicos da Água , Animais , Aquicultura , Catalase/metabolismo , Desinfetantes/toxicidade , Ecossistema , Glutationa Transferase/metabolismo , Estresse Oxidativo , Propilenoglicóis , Superóxido Dismutase/metabolismo , Poluentes Químicos da Água/toxicidade
5.
Turk Neurosurg ; 31(3): 447-459, 2021.
Artigo em Inglês | MEDLINE | ID: mdl-33978213

RESUMO

AIM: To analyze the Glutathione S-transferase (GST)-P, GST-M, cytochrome p450 (CYP)1-A1, CYP1-B1, and multidrug resistance (MDR)-1 expressions in malignant intracranial tumor (ICT)s, and to elicit their role on patient survival. MATERIAL AND METHODS: GST-P, GST-M, CYP1-A1, CYP1-B1, and MDR-1 expressions were analyzed using immunostaining in 149 samples from 141 patients with preoperative ICT diagnosis. The case characteristics were reviewed, and the enzyme expressions were equated based on the age, gender, and tumor type. Then, 77 of 141 patients with malignant ICT and complete medical records postoperative were also investigated in detail for the relationship between the diagnosis, enzyme expression, and overall survival. RESULTS: The average age was 49.44 years, with 83 (58.45%) male patients. Among the 77 malignant ICTs, 38 (49.3%) and 29 were glial tumors and metastases, respectively, with a 13.35-month overall survival. Patients with metastatic tumor have approximately threefold higher GSTP level than those with glial tumors. MDR-1 expression was approximately twofold higher in > 60-year-old patients. No statistically significant association was found between patients? smoking behaviors, alcohol consumption, and overall survival. Only MDR-1 expression was correlated with overall survival. Better overall survival was observed in patients with a negative MDR-1 expression than those with a positive one. CONCLUSION: MDR-1 is an important indicator of survival in malignant intracranial tumor patients. Longer survival is associated with negative MDR-1 expression.


Assuntos
Membro 1 da Subfamília B de Cassetes de Ligação de ATP/metabolismo , Neoplasias Encefálicas/tratamento farmacológico , Sistema Enzimático do Citocromo P-450/metabolismo , Resistencia a Medicamentos Antineoplásicos/fisiologia , Glutationa Transferase/metabolismo , Adolescente , Adulto , Idoso , Idoso de 80 Anos ou mais , Neoplasias Encefálicas/metabolismo , Neoplasias Encefálicas/mortalidade , Criança , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Taxa de Sobrevida , Adulto Jovem
6.
Toxins (Basel) ; 13(3)2021 03 22.
Artigo em Inglês | MEDLINE | ID: mdl-33809820

RESUMO

Concerns about resistance development to conventional insecticides in diamondback moth (DBM) Plutella xylostella (L.), the most destructive pest of Brassica vegetables, have stimulated interest in alternative pest management strategies. The toxicity of Bacillus thuringiensis subsp. aizawai (Bt GO33A) combined with chlorantraniliprole (Chl) has not been documented. Here, we examined single and combined toxicity of chlorantraniliprole and Bt to assess the levels of resistance in four DBM strains. Additionally, enzyme activities were tested in field-original highly resistant (FOH-DBM), Bt-resistant (Bt-DBM), chlorantraniliprole-resistant (CL-DBM), and Bt + chlorantraniliprole-resistant (BtC-DBM) strains. The Bt product had the highest toxicity to all four DBM strains followed by the mixture of insecticides (Bt + Chl) and chlorantraniliprole. Synergism between Bt and chlorantraniliprole was observed; the combination of Bt + (Bt + Chl) (1:1, LC50:LC50) was the most toxic, showing a synergistic effect against all four DBM strains with a poison ratio of 1.35, 1.29, 1.27, and 1.25. Glutathione S-transferase (GST) and carboxyl-esterase (CarE) activities showed positive correlations with chlorantraniliprole resistance, but no correlation was observed with resistance to Bt and Bt + Chl insecticides. Expression of genes coding for PxGST, CarE, AChE, and MFO using qRT-PCR showed that the PxGST and MFO were significantly overexpressed in Bt-DBM. However, AChE and CarE showed no difference in the four DBM strains. Mixtures of Bt with chlorantraniliprole exhibited synergistic effects and may aid the design of new combinations of pesticides to delay resistance in DBM strains substantially.


Assuntos
Bacillus thuringiensis/metabolismo , Brassica/parasitologia , Resistência a Inseticidas , Inseticidas/farmacologia , Mariposas/efeitos dos fármacos , Mariposas/microbiologia , Controle Biológico de Vetores , ortoaminobenzoatos/farmacologia , Acetilcolinesterase/genética , Acetilcolinesterase/metabolismo , Animais , Bacillus thuringiensis/genética , Carboxilesterase/genética , Carboxilesterase/metabolismo , Sistema Enzimático do Citocromo P-450/genética , Sistema Enzimático do Citocromo P-450/metabolismo , Regulação Enzimológica da Expressão Gênica , Glutationa Transferase/genética , Glutationa Transferase/metabolismo , Resistência a Inseticidas/genética , Mariposas/enzimologia , Mariposas/genética
7.
Mol Biol Rep ; 48(3): 2591-2599, 2021 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-33791906

RESUMO

Bisphenol F (BPF) has been used frequently in the plastics industry and the production of daily consumer products as an alternative to bisphenol A (BPA). It was aimed herein to determine the cytotoxic effects of BPF on hepatocytes isolated from the liver of rainbow trout (Oncorhyncus mykiss) using lactate dehydrogenase (LDH) assay and antioxidant defence system indicators. The cultured hepatocytes were exposed to seven concentrations (0, 15.63, 31.25, 62.50, 125, 250, and 500 µM) of BPF for 24 h. According to the LDH assay, the percentage of cytotoxicity was increased dose dependently in the cells. The malondialdehyde content, which is indicative of lipid peroxidation, was increased significantly at BPF concentrations between 15.63 and 250 µM, whereas it remained unchanged with a concentration of 500 µM. The activities of superoxide dismutase were increased, while those of catalase were decreased with all of the BPF concentrations. Elevated levels of reduced glutathione content were determined with BPF concentrations between 15.63 and 250 µM, but decreased significantly with a concentration of 500 µM. Significant increases in the activities of the glutathione peroxidase were found in hepatocytes treated with BPF at concentrations of 31.25 to 500 µM. GST activity was only significantly increased with a BPF concentration of 250 µM. The results showed that the toxic mechanism of BPF was mainly based on cell membrane damage and oxidative stress, which have an influence on antioxidant defences. Therefore, BPF should be reconsidered as a safe alternative instead of BPA in the manufacturing of industrial or daily products.


Assuntos
Antioxidantes/metabolismo , Compostos Benzidrílicos/toxicidade , Separação Celular , Hepatócitos/metabolismo , Oncorhynchus mykiss/metabolismo , Fenóis/toxicidade , Animais , Catalase/metabolismo , Morte Celular/efeitos dos fármacos , Células Cultivadas , Feminino , Glutationa/metabolismo , Glutationa Peroxidase/metabolismo , Glutationa Transferase/metabolismo , Hepatócitos/efeitos dos fármacos , Malondialdeído/metabolismo , Superóxido Dismutase/metabolismo
8.
Molecules ; 26(8)2021 Apr 16.
Artigo em Inglês | MEDLINE | ID: mdl-33923444

RESUMO

PACRG (Parkin co-regulated gene) shares a bi-directional promoter with the Parkinson's disease-associated gene Parkin, but the physiological roles of PACRG have not yet been fully elucidated. Recombinant expression methods are indispensable for protein structural and functional studies. In this study, the coding region of PACRG was cloned to a conventional vector pQE80L, as well as two cold-shock vectors pCold II and pCold-GST, respectively. The constructs were transformed into Escherichia coli (DE3), and the target proteins were overexpressed. The results showed that the cold-shock vectors are more suitable for PACRG expression. The soluble recombinant proteins were purified with Ni2+ chelating column, glutathione S-transferase (GST) affinity chromatography and gel filtration. His6 pull down assay and LC-MS/MS were carried out for identification of PACRG-binding proteins in HEK293T cell lysates, and a total number of 74 proteins were identified as potential interaction partners of PACRG. GO (Gene ontology) enrichment analysis (FunRich) of the 74 proteins revealed multiple molecular functions and biological processes. The highest proportion of the 74 proteins functioned as transcription regulator and transcription factor activity, suggesting that PACRG may play important roles in regulation of gene transcription.


Assuntos
Glutationa Transferase/metabolismo , Cromatografia de Afinidade , Cromatografia em Gel , Glutationa Transferase/isolamento & purificação , Células HEK293 , Humanos , Proteínas dos Microfilamentos/isolamento & purificação , Proteínas dos Microfilamentos/metabolismo , Chaperonas Moleculares/isolamento & purificação , Chaperonas Moleculares/metabolismo , Ligação Proteica , Espectrometria de Massas em Tandem , Ubiquitina-Proteína Ligases/metabolismo
9.
Int J Mol Sci ; 22(6)2021 Mar 17.
Artigo em Inglês | MEDLINE | ID: mdl-33802702

RESUMO

Our previous study demonstrated that the glutathione S-transferase Mu 5 (GSTM5) gene is highly CpG-methylated in bladder cancer cells and that demethylation by 5-aza-dC activates GSTM5 gene expression. The aim of the present study was to investigate the role of GSTM5 in bladder cancer. The levels of GSTM5 gene expression and DNA methylation were analyzed in patients with bladder cancer, and functional studies of GSTM5 were conducted using GSTM5 overexpression in cultured bladder cancer cells. Clinical analysis revealed that the GSTM5 mRNA expression was lower in bladder cancer tissues than in normal tissues and that the level of GSTM5 DNA methylation was higher in bladder cancer tissues than in normal urine pellets. Overexpression of GSTM5 decreased cell proliferation, migration and colony formation capacity. Glutathione (GSH) assay results indicated that cellular GSH concentration was decreased by GSTM5 expression and that GSH supplementation reversed the decrease in proliferation and migration of cells overexpressing GSTM5. By contrast, a GSH synthesis inhibitor significantly decreased 5637 cell GSH levels, survival and migration. Furthermore, GSTM5 overexpression inhibited the adhesion of cells to the extracellular matrix protein fibronectin. To elucidate the effect of GSTM5 on anticancer drugs used to treat bladder cancer, cellular viability was compared between cells with or without GSTM5 overexpression. GSTM5-overexpressed cells showed no significant change in the cytotoxicity of cisplatin or mitomycin C in 5637, RT4 and BFTC 905 cells. Though a degree of resistance to doxorubicin was noted in 5637 cells overexpressing GSTM5, no such resistance was observed in RT4 and BFTC 905 cells. In summary, GSTM5 plays a tumor suppressor role in bladder cancer cells without significantly affecting chemoresistance to cisplatin and mitomycin C, and the cellular GSH levels highlight a key mechanism underlying the cancer inhibition effect of GSTM5. These findings suggest that low gene expression and high DNA methylation levels of GSTM5 may act as tumor markers for bladder cancer.


Assuntos
Antineoplásicos/metabolismo , Biomarcadores Tumorais/metabolismo , Glutationa Transferase/metabolismo , Neoplasias da Bexiga Urinária/enzimologia , Adulto , Idoso , Idoso de 80 Anos ou mais , Butionina Sulfoximina/farmacologia , Adesão Celular/efeitos dos fármacos , Morte Celular/efeitos dos fármacos , Linhagem Celular Tumoral , Movimento Celular/efeitos dos fármacos , Movimento Celular/genética , Proliferação de Células/efeitos dos fármacos , Proliferação de Células/genética , Cisplatino/farmacologia , Metilação de DNA/efeitos dos fármacos , Metilação de DNA/genética , Regulação para Baixo/efeitos dos fármacos , Regulação para Baixo/genética , Doxorrubicina/farmacologia , Feminino , Regulação Neoplásica da Expressão Gênica/efeitos dos fármacos , Glutationa/metabolismo , Glutationa Transferase/genética , Humanos , Masculino , Pessoa de Meia-Idade , Mitomicina/farmacologia , Regiões Promotoras Genéticas/genética , RNA Mensageiro/genética , RNA Mensageiro/metabolismo , Caracteres Sexuais , Neoplasias da Bexiga Urinária/genética
10.
J Enzyme Inhib Med Chem ; 36(1): 885-894, 2021 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-33752574

RESUMO

Here we investigated the effects of different levels of royal jelly in zebrafish (Danio rerio) diets [0.0% (D1); 0.1% (D2); 0.4% (D3); 1.6% (D4) vs 6.4% (D5)] on the activity and expression profiles of superoxide dismutase, catalase, glutathione reductase, glutathione peroxidase and glutathione S-transferase. Muscle, liver and kidney tissue samples were obtained from fish fed during 8 weeks. In these tissues, enzyme activity was determined by means of spectrophotometer and gene expression by quantitative real-time PCR. mRNA levels of the enzymes were elevated in almost all diet groups compared to the control (D1). It was determined that enzyme activities were also increased in general by supplementation of royal jelly although some decreases were also observed. However, the significant correlation between gene expression and enzyme activity was not observed in all tissues. It was concluded that main regulation occurs with post-translational modifications although effects at transcriptomic level demonstrated a snap variation.


Assuntos
Catalase/genética , Ácidos Graxos/farmacologia , Glutationa Peroxidase/genética , Glutationa Redutase/genética , Glutationa Transferase/genética , Estresse Oxidativo/efeitos dos fármacos , Superóxido Dismutase/genética , Peixe-Zebra , Animais , Catalase/análise , Catalase/metabolismo , Dieta , Ácidos Graxos/administração & dosagem , Perfilação da Expressão Gênica , Glutationa Peroxidase/análise , Glutationa Peroxidase/metabolismo , Glutationa Redutase/análise , Glutationa Redutase/metabolismo , Glutationa Transferase/análise , Glutationa Transferase/metabolismo , Estresse Oxidativo/genética , Processamento de Proteína Pós-Traducional , RNA Mensageiro/análise , RNA Mensageiro/genética , RNA Mensageiro/metabolismo , Reação em Cadeia da Polimerase em Tempo Real , Espectrofotometria , Superóxido Dismutase/análise , Superóxido Dismutase/metabolismo
11.
Plant Sci ; 305: 110827, 2021 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-33691961

RESUMO

Anthocyanins are flavonoid pigments providing plants a range of colors from red, pink, orange to blue. Anthocyanins are synthesized in the cytosol but accumulate predominantly in the vacuoles through vacuolar sequestration involving glutathione S-transferases (GSTs) and multidrug and toxic compound extrusion (MATE) and the ATP binding cassette (ABC) transporters. However, little is known about anthocyanin-related GSTs in Upland cotton (Gossypium hirsutum L.). In this study, we performed genome-wide identification of GST genes in Upland cotton and identified GST genes functioning in accumulation of anthocyanins. We demonstrated that GhGSTF12 was able to complement the defective leaf color phenotypes of the Arabidopsis tt19 mutant caused by mutation in a GSTF gene. Virus-induced silencing of GhGSTF12 in the red leaf cultivar turned its red color to green and transient overexpression of GhGSTF12 accelerated anthocyanin accumulation in the red leaf cultivar but not in the green leaf cultivar. Collectively, GhGSTF12 may be involved in transport of anthocyanins from cytosol to vacuoles in cotton. These results also demonstrated a conserved function of plant GSTF genes in anthocyanin accumulation and provide a candidate gene for manipulating pigmentation in cotton tissues.


Assuntos
Antocianinas/biossíntese , Antocianinas/genética , Glutationa Transferase/genética , Glutationa Transferase/metabolismo , Gossypium/genética , Gossypium/metabolismo , Pigmentação/genética , Pigmentação/fisiologia , Produtos Agrícolas/genética , Produtos Agrícolas/metabolismo , Regulação da Expressão Gênica de Plantas , Genes de Plantas , Variação Genética , Genótipo , Filogenia
12.
Chemosphere ; 275: 130000, 2021 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-33667769

RESUMO

Lithobates catesbeianus tadpoles were exposed for 96 h to water from two sites of the Sorocaba River (summer and winter), Ibiúna (PI) and Itupararanga reservoir (PIR) that contained metals. In the liver, in PI, the glutathione peroxidase (GPx) decreased, and the glutathione S-transferase (GST) and carbonyl proteins (PCO) increased. In PIR, the glutathione reduced (GSH) increased, while there was a decrease in catalase (CAT), GPx, GST, PCO, and superoxide dismutase (SOD). In winter, GPx and GST increased in both points. Regarding the kidneys, lipoperoxidation (LPO) levels and GST decreased, while GSH increased in the summer. In the winter, LPO increased in PI. In the muscle, in the summer, there was an increase in GSH and GST and change in PCO. In the winter, the levels of PCO increased and CAT decreased in PIR. The area and volume of the hepatocyte and nucleus area increased in the summer and decreased in the winter. Hepatic melanin decreased in the summer after exposure to PIR water. There were the systemic effects of Sorocaba River water exposure at different times of the year with alterations in biomarkers at different levels, in which kidney shows highest Integrated Response of Biomarkers (IBR) value followed by liver and muscle. Biochemical biomarkers were more sensitive than morphological ones. The more sensitive biochemical markers were MT, PCO, GST and LPO. These effects confirm the hypothesis of metabolic alteration in bullfrog tadpoles by the Sorocaba River water.


Assuntos
Rios , Poluentes Químicos da Água , Animais , Biomarcadores/metabolismo , Catalase/metabolismo , Glutationa/metabolismo , Glutationa Peroxidase/metabolismo , Glutationa Transferase/metabolismo , Larva/metabolismo , Peroxidação de Lipídeos , Estresse Oxidativo , Rana catesbeiana/metabolismo , Superóxido Dismutase/metabolismo , Água , Poluentes Químicos da Água/toxicidade
13.
Nat Commun ; 12(1): 1728, 2021 03 19.
Artigo em Inglês | MEDLINE | ID: mdl-33741927

RESUMO

Microsomal glutathione S-transferase 2 (MGST2) produces leukotriene C4, key for intracrine signaling of endoplasmic reticulum (ER) stress, oxidative DNA damage and cell death. MGST2 trimer restricts catalysis to only one out of three active sites at a time, but the molecular basis is unknown. Here, we present crystal structures of human MGST2 combined with biochemical and computational evidence for a concerted mechanism, involving local unfolding coupled to global conformational changes that regulate catalysis. Furthermore, synchronized changes in the biconical central pore modulate the hydrophobicity and control solvent influx to optimize reaction conditions at the active site. These unique mechanistic insights pertain to other, structurally related, drug targets.


Assuntos
Glutationa Transferase/química , Glutationa Transferase/genética , Glutationa Transferase/metabolismo , Sítios de Ligação , Catálise , Domínio Catalítico , Cristalografia por Raios X , Retículo Endoplasmático/metabolismo , Humanos , Leucotrieno C4/metabolismo , Simulação de Dinâmica Molecular , Mutagênese Sítio-Dirigida , Estresse Oxidativo , Conformação Proteica
14.
Arch Insect Biochem Physiol ; 106(4): e21770, 2021 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-33660279

RESUMO

MGST2 is a member of the MAPEG superfamily, which participates in LTC4 synthesis and plays important roles in the regulation of the oxidative stress pathway and some diseases. Here, we isolated a previously uncharacterized gene in Apis cerana cerana named AccMGST2 by reverse transcription-polymerase chain reaction. The biological characteristics of AccMGST2 were analyzed by bioinformatics. The amino acid sequence similarity between AccMGST2 and AmMGST2 of Apis mellifera reached 96.08%. The expression characteristics of AccMGST2 were explored in several tissues. The quantitative real-time polymerase chain reaction results showed that the AccMGST2 gene was highly expressed in the head and muscle and that AccMGST2 expression responded to oxidative stress caused by different abiotic stresses. AccMGST2 was silenced using RNA interference, which decreased the expression levels of some MAPK and antioxidant genes. Therefore, we conclude that AccMGST2 is involved in the regulation of oxidative stress in A. cerana cerana.


Assuntos
Abelhas/genética , Glutationa Transferase , Sequência de Aminoácidos , Animais , Antioxidantes/metabolismo , Genes de Insetos , Glutationa Transferase/química , Glutationa Transferase/genética , Glutationa Transferase/metabolismo , Proteínas de Insetos/química , Proteínas de Insetos/genética , Proteínas de Insetos/metabolismo , Estresse Oxidativo/genética , Filogenia
15.
Free Radic Biol Med ; 165: 184-190, 2021 03.
Artigo em Inglês | MEDLINE | ID: covidwho-1056615

RESUMO

Several recent reviews have suggested a role of oxidative stress in the pathophysiology of COVID-19, but its interplay with disease severity has not been revealed yet. In the present study, we aimed to investigate the association between the severity of COVID-19 and oxidative stress parameters. Clinical data of 77 patients with COVID-19 admitted to the hospital were analyzed and divided into moderate (n = 44) and severe (n = 33) groups based on their clinical condition. Production of oxidant (hydrogen peroxide) and defense antioxidants (total antioxidant capacity, reduced and oxidized glutathione, glutathione s-transferase), and oxidative damage (malondialdehyde, carbonyl, and sulfhydryl) were assessed using the serum samples. The results revealed that severe patients who presented high serum leukocyte count and CRP level stayed for a longer period in the hospital. However, there was no correlation observed between the oxidative stress parameters and degree of COVID-19 severity in the present study. In conclusion, these results indicate that the disease severity may not be a detrimental factor contributing to the changes in the redox profile of hospitalized patients with COVID-19.


Assuntos
COVID-19/metabolismo , Estresse Oxidativo/fisiologia , SARS-CoV-2/fisiologia , Adulto , Idoso , Brasil/epidemiologia , COVID-19/epidemiologia , Estudos de Coortes , Progressão da Doença , Suscetibilidade a Doenças , Feminino , Glutationa/metabolismo , Glutationa Transferase/metabolismo , Humanos , Peróxido de Hidrogênio/metabolismo , Masculino , Pessoa de Meia-Idade , Estudos Prospectivos
16.
Free Radic Biol Med ; 165: 184-190, 2021 03.
Artigo em Inglês | MEDLINE | ID: mdl-33524532

RESUMO

Several recent reviews have suggested a role of oxidative stress in the pathophysiology of COVID-19, but its interplay with disease severity has not been revealed yet. In the present study, we aimed to investigate the association between the severity of COVID-19 and oxidative stress parameters. Clinical data of 77 patients with COVID-19 admitted to the hospital were analyzed and divided into moderate (n = 44) and severe (n = 33) groups based on their clinical condition. Production of oxidant (hydrogen peroxide) and defense antioxidants (total antioxidant capacity, reduced and oxidized glutathione, glutathione s-transferase), and oxidative damage (malondialdehyde, carbonyl, and sulfhydryl) were assessed using the serum samples. The results revealed that severe patients who presented high serum leukocyte count and CRP level stayed for a longer period in the hospital. However, there was no correlation observed between the oxidative stress parameters and degree of COVID-19 severity in the present study. In conclusion, these results indicate that the disease severity may not be a detrimental factor contributing to the changes in the redox profile of hospitalized patients with COVID-19.


Assuntos
COVID-19/metabolismo , Estresse Oxidativo/fisiologia , SARS-CoV-2/fisiologia , Adulto , Idoso , Brasil/epidemiologia , COVID-19/epidemiologia , Estudos de Coortes , Progressão da Doença , Suscetibilidade a Doenças , Feminino , Glutationa/metabolismo , Glutationa Transferase/metabolismo , Humanos , Peróxido de Hidrogênio/metabolismo , Masculino , Pessoa de Meia-Idade , Estudos Prospectivos
17.
Sci Total Environ ; 772: 145034, 2021 Jun 10.
Artigo em Inglês | MEDLINE | ID: mdl-33571776

RESUMO

Fish can be highly vulnerable to environmental pressures because they are exposed to oxidative stressors in the aquatic environment. Such stressors can affect the levels of antioxidant biomarkers against reactive oxygen species (ROS). With this study we investigated the oxidative stress ecology in Danube barbel (Barbus balcanicus) from the Barbucina creek (northeast Italy), a watercourse in the Collio winegrowing district. To do this, superoxide dismutase (SOD), catalase (CAT), and glutathione S-transferase (GST) activity was measured in gills, liver, and muscle, while metallothioneins (MT) and trace and rare earth elements (REEs) levels were determined in muscle. The effect of environmental factors (physicochemical parameters of water, trace elements and REEs) on oxidative stress biomarkers was thus assessed. High concentrations were determined for cerium (Ce), scandium (Sc), neodymium (Nd), lanthanum (La), yttrium (Y), and praseodymium (Pr) among the REEs. Among the trace elements, arsenic (As), copper (Cu), and mercury (Hg) levels were higher compared to published data, suggesting their role as stressors. The multiple linear regression (MLR) model showed a statistically significant association (R2 = 0.858; F = 10.07; p = 0.015) between As, Cu, Hg, and Pr and SOD activity in the gills, indicating a functional relationship between them. Differently, CAT activity was significantly higher in the liver, probably in response to long-term Cu contamination of the watercourse. This was confirmed by the MLR model that showed a significant association (R2 = 0.638; F = 8.152; p = 0.02) between the concentration of MT and of Cu. Our data show a biochemical defensive response by Danube barbel to the disturbances in the aquatic ecosystem of the Barbucina creek. These insights advance our understanding of the role and the effects of environmental factors as trace elements and REEs on oxidative stress in fish.


Assuntos
Cyprinidae , Poluentes Químicos da Água , Animais , Biomarcadores/metabolismo , Catalase/metabolismo , Cyprinidae/metabolismo , Ecossistema , Brânquias/metabolismo , Glutationa Transferase/metabolismo , Itália , Estresse Oxidativo , Superóxido Dismutase/metabolismo , Água , Poluentes Químicos da Água/análise , Poluentes Químicos da Água/toxicidade
18.
Oxid Med Cell Longev ; 2021: 6678924, 2021.
Artigo em Inglês | MEDLINE | ID: mdl-33574979

RESUMO

Deletion polymorphism of glutathione S-transferase M1 (GSTM1), a phase II detoxification and antioxidant enzyme, increases susceptibility to end-stage renal disease (ESRD) as well as the development of cardiovascular diseases (CVD) among ESRD patients and leads to their shorter cardiovascular survival. The mechanisms by which GSTM1 downregulation contributes to oxidative stress and inflammation in endothelial cells in uremic conditions have not been investigated so far. Therefore, the aim of the present study was to elucidate the effects of GSTM1 knockdown on oxidative stress and expression of a panel of inflammatory markers in human umbilical vein endothelial cells (HUVECs) exposed to uremic serum. Additionally, we aimed to discern whether GSTM1-null genotype is associated with serum levels of adhesion molecules in ESRD patients. HUVECs treated with uremic serum exhibited impaired redox balance characterized by enhanced lipid peroxidation and decreased antioxidant enzyme activities, independently of the GSTM1 knockdown. In response to uremic injury, HUVECs exhibited alteration in the expression of a series of inflammatory cytokines including retinol-binding protein 4 (RBP4), regulated on activation, normal T cell expressed and secreted (RANTES), C-reactive protein (CRP), angiogenin, dickkopf-1 (Dkk-1), and platelet factor 4 (PF4). GSTM1 knockdown in HUVECs showed upregulation of monocyte chemoattractant protein-1 (MCP-1), a cytokine involved in the regulation of monocyte migration and adhesion. These cells also have shown upregulated intracellular and vascular cell adhesion molecules (ICAM-1 and VCAM-1). In accordance with these findings, the levels of serum ICAM-1 and VCAM-1 (sICAM-1 and sVCAM-1) were increased in ESRD patients lacking GSTM1, in comparison with patients with the GSTM1-active genotype. Based on these results, it may be concluded that incubation of endothelial cells in uremic serum induces redox imbalance accompanied with altered expression of a series of cytokines involved in arteriosclerosis and atherosclerosis. The association of GSTM1 downregulation with the altered expression of adhesion molecules might be at least partly responsible for the increased susceptibility of ESRD patients to CVD.


Assuntos
Glutationa Transferase/metabolismo , Células Endoteliais da Veia Umbilical Humana/metabolismo , Molécula 1 de Adesão Intercelular/metabolismo , Uremia/metabolismo , Molécula 1 de Adesão de Célula Vascular/metabolismo , Biomarcadores/metabolismo , Citocinas/metabolismo , Deleção de Genes , Glutationa Peroxidase/metabolismo , Humanos , Falência Renal Crônica/sangue , Falência Renal Crônica/metabolismo , Malondialdeído/metabolismo , Estresse Oxidativo , Proteoma/metabolismo , Espécies Reativas de Oxigênio/metabolismo , Superóxido Dismutase/metabolismo , Uremia/sangue
19.
FASEB J ; 35(2): e21352, 2021 02.
Artigo em Inglês | MEDLINE | ID: mdl-33543805

RESUMO

Toxoplasma gondii is an apicomplexan parasite, which has three unique secretory organelles: micronemes, rhoptries, and dense granules. Almost all the secreted proteins are transported through the endoplasmic reticulum (ER) and Golgi system to function in their respective destination by accurate targeting and packaging. Glutathione S-transferase (GST) is a supergene family enzyme that has multiple functions, which include regulation of cell proliferation and death signaling pathways, and participation in transportation and metabolism in mammal cells. However, the role of GST in Toxoplasma gondii has not been explained. In this study, we identified three GST proteins in T gondii, of which GST2 acts as a membrane protein that localizes to the Golgi-endosomal system and colocalizes with proteins involved in vesicle transport as well, including synaptobrevin, putative sortilin (VPS10), Rab5 and Rab6, which function as vesicle transport factors. Moreover, the loss of TgGST2 leads to Rab5 and Rab6 distribution of discrete puncta, and incorrect localization and decreased expression of several secretory proteins, and to significantly reduced invasion capacity and virulence to mice. Consistent with its relation to vesicle transport proteins, the distribution of TgGST2 relies on post-Golgi trafficking. Overall, our findings demonstrated that TgGST2 contributes to vesicle trafficking and plays a critical role in parasite lytic cycle.


Assuntos
Glutationa Transferase/metabolismo , Proteínas de Protozoários/metabolismo , Via Secretória , Toxoplasma/metabolismo , Proteínas Adaptadoras de Transporte Vesicular/metabolismo , Endossomos/metabolismo , Complexo de Golgi/metabolismo , Transporte Proteico , Proteínas R-SNARE/metabolismo , Proteínas rab de Ligação ao GTP/metabolismo , Proteínas rab5 de Ligação ao GTP/metabolismo
20.
Plant Sci ; 304: 110811, 2021 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-33568308

RESUMO

Maize (Zea mays L.) production is severely affected by northern corn leaf blight (NCLB), which is a destructive foliar disease caused by Setosphaeria turcica. In recent years, studies on the interaction between maize and S. turcica have been focused at the transcription level, with no research yet at the protein level. Here, we applied tandem mass tag labelling and liquid chromatography-tandem mass spectrometry to investigate the proteomes of maize leaves at 24 h and 72 h post-inoculation (hpi) with S. turcica. In total, 4740 proteins encoded by 4711 genes were quantified in this study. Clustering analyses provided an understanding of the dynamic reprogramming of leaves proteomes by revealing the functions of different proteins during S. turcica infection. Screening and classification of differentially expressed proteins (DEPs) revealed that numerous defense-related proteins, including defense marker proteins and proteins related to the phenylpropanoid lignin biosynthesis, benzoxazine biosynthesis and the jasmonic acid signalling pathway, participated in the defense responses of maize to S. turcica infection. Furthermore, the earlier induction of GST family proteins contributed to the resistance to S. turcica. In addition, the protein-protein interaction network of DEPs suggests that some defense-related proteins, for example, ZmGEB1, a hub node, play key roles in defense responses against S. turcica infection. Our study findings provide insight into the complex responses triggered by S. turcica at the protein level and lay the foundation for studying the interaction process between maize and S. turcica infection.


Assuntos
Ascomicetos , Doenças das Plantas/microbiologia , Zea mays/microbiologia , Ciclopentanos/metabolismo , Cromatografia Gasosa-Espectrometria de Massas , Glutationa Transferase/metabolismo , Redes e Vias Metabólicas , Oxilipinas/metabolismo , Doenças das Plantas/imunologia , Reguladores de Crescimento de Plantas/metabolismo , Folhas de Planta/imunologia , Folhas de Planta/metabolismo , Folhas de Planta/microbiologia , Proteínas de Plantas/metabolismo , Mapas de Interação de Proteínas , Proteômica , Transcriptoma , Zea mays/imunologia , Zea mays/metabolismo
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