THE EFFECT OF INTERMITTENT HYPOXIC TRAINING UNDER OXIDATIVE STRESS PARAMETERS IN WISTAR RATS FED ON STANDARD AND HIGH FAT DIET

Considerando-se que dietas ricas em gordura levam ao estresse oxidativo, causando lesões nas células e que o treinamento hipóxico intermitente (THI) aumenta as defesas antioxidantes endógenas em diversas situações, o objetivo deste estudo foi avaliar o efeito do THI em parâmetros de estresse oxidativo e defesas antioxidantes em fígado de ratos Wistar alimentados com dieta rica em gordura e/ou dieta padrão. Ratos Wistar foram divididos em grupos alimentados com dieta padrão ou rica em gordura. Os grupos foram submetidos a hipóxia intermitente (HI), 15 minutos HI (14-11% O2) intercalados com cinco minutos de re-oxigenação ou sessões de normóxia (N) (21% O2), por um período de duas horas diárias durante 30 dias. Os ratos Wistar alimentados com dieta padrão, e submetidas a HI, apresentaram uma redução de 37,7% na concentração de substâncias reativas ao ácido tiobarbitúrico (TBARS) e aumento de 34,66% e 39,8% no conteúdo de superóxido dismutase (SOD) e catalase (CAT), respectivamente, em comparação com o seu controlo (normoxia). No grupo com dieta rica em gordura, não houve diferença estatística entre os subgrupos HI e N. Nossos dados, que demonstram que o THI possui efeito antioxidante no fígado de ratos Wistar, argumentam em favor do uso alternativo de protocolos de hipoxia intermitente no tratamento de determinadas patologias

TRP14 is the rate-limiting enzyme for intracellular cystine reduction and regulates proteome cysteinylation

It has remained unknown how cells reduce cystine taken up from the extracellular space, which is a required step for further utilization of cysteine in key processes such as protein or glutathione synthesis. Here, we show that the thioredoxin-related protein of 14 kDa (TRP14, encoded by TXNDC17) is the rate-limiting enzyme for intracellular cystine reduction. When TRP14 is genetically knocked out, cysteine synthesis through the transsulfuration pathway becomes the major source of cysteine in human cells, and knockout of both pathways becomes lethal in C. elegans subjected to proteotoxic stress. TRP14 can also reduce cysteinyl moieties on proteins, rescuing their activities as here shown with cysteinylated peroxiredoxin 2. Txndc17 knockout mice were, surprisingly, protected in an acute pancreatitis model, concomitant with activation of Nrf2-driven antioxidant pathways and upregulation of transsulfuration. We conclude that TRP14 is the evolutionarily conserved enzyme principally responsible for intracellular cystine reduction in C. elegans, mice, and humans.RED2018-102576-T/MEC | Agencia Estatal de Investigación (AEI) PID2019-108615RB-I00/MEC | Agencia Estatal de Investigación (AEI) AG040020/United States National Institutes of Health P40 OD010440/OD/NIH HHS/United States ED_18-1-2019-0025/The National Research. Development and Innovation Offic

Hepatite B: percepção de risco e adoção de medidas de biossegurança por manicures/pedicures de Itaúna-MG

Trata-se de um estudo quantitativo, transversal, exploratrio, descritivo e observacional que teve como objetivo analisar a percepo de risco de contaminao com o vrus da hepatite B (HBV) e a adoo de medidas de biossegurana dos manicures e pedicures do municpio de Itana-MG . A pesquisa foi realizada partindo-se do nmero total de sales de beleza que possuem alvar de funcionamento junto a Secretaria Municipal de Sade. O clculo amostral forneceu um nmero de 127 estabelecimentos que foram selecionados de forma aleatria atravs de sorteio, onde apenas um profissional de cada estabelecimento participou da pesquisa. Os dados foram obtidos atravs de questionrio estruturado e os resultados mostraram que 79,5% dos profissionais afirmaram conhecer as formas de transmisso do HBV, contudo, houve considervel falta de adeso ao uso de luvas descartveis (87,4%) e apenas 1,1% dos profissionais que utilizavam estufa seguiram a relao adequada de tempo e temperatura necessria para esterilizao efetiva. Percebeu-se um desconhecimento sobre a importncia do esquema vacinal completo para a hepatite B. Conclumos que a percepo de risco dos manicures e pedicures com relao contaminao com o HBV falha, evidenciando a necessidade da elaborao de cursos de capacitao que promovam uma maior conscientizao sobre biossegurana nestes profissionais

Redox signaling in the gastrointestinal tract

Redox signaling regulates physiological self-renewal, proliferation, migration and differentiation in gastrointestinal epithelium by modulating Wnt/β-catenin and Notch signaling pathways mainly through NADPH oxidases (NOXs). In the intestine, intracellular and extracellular thiol redox status modulates the proliferative potential of epithelial cells. Furthermore, commensal bacteria contribute to intestine epithelial homeostasis through NOX1- and dual oxidase 2-derived reactive oxygen species (ROS). The loss of redox homeostasis is involved in the pathogenesis and development of a wide diversity of gastrointestinal disorders, such as Barrett's esophagus, esophageal adenocarcinoma, peptic ulcer, gastric cancer, ischemic intestinal injury, celiac disease, inflammatory bowel disease and colorectal cancer. The overproduction of superoxide anion together with inactivation of superoxide dismutase are involved in the pathogenesis of Barrett's esophagus and its transformation to adenocarcinoma. In Helicobacter pylori-induced peptic ulcer, oxidative stress derived from the leukocyte infiltrate and NOX1 aggravates mucosal damage, especially in HspB+ strains that downregulate Nrf2. In celiac disease, oxidative stress mediates most of the cytotoxic effects induced by gluten peptides and increases transglutaminase levels, whereas nitrosative stress contributes to the impairment of tight junctions. Progression of inflammatory bowel disease relies on the balance between pro-inflammatory redox-sensitive pathways, such as NLRP3 inflammasome and NF-κB, and the adaptive up-regulation of Mn superoxide dismutase and glutathione peroxidase 2. In colorectal cancer, redox signaling exhibits two Janus faces: On the one hand, NOX1 up-regulation and derived hydrogen peroxide enhance Wnt/β-catenin and Notch proliferating pathways; on the other hand, ROS may disrupt tumor progression through different pro-apoptotic mechanisms. In conclusion, redox signaling plays a critical role in the physiology and pathophysiology of gastrointestinal tract

Obesity causes PGC-1α deficiency in the pancreas leading to marked IL-6 upregulation via NF-κB in acute pancreatitis

Obesity is associated with local and systemic complications in acute pancreatitis. PPARγ coactivator 1α (PGC-1α) is a transcriptional coactivator and master regulator of mitochondrial biogenesis that exhibits dysregulation in obese subjects. Our aims were: (1) to study PGC-1α levels in pancreas from lean or obese rats and mice with acute pancreatitis; and (2) to determine the role of PGC-1α in the inflammatory response during acute pancreatitis elucidating the signaling pathways regulated by PGC-1α. Lean and obese Zucker rats and lean and obese C57BL6 mice were used first; subsequently, wild-type and PGC-1α knockout (KO) mice with cerulein-induced pancreatitis were used to assess the inflammatory response and expression of target genes. Ppargc1a mRNA and protein levels were markedly downregulated in pancreas of obese rats and mice versus lean animals. PGC-1α protein levels increased in pancreas of lean mice with acute pancreatitis, but not in obese mice with pancreatitis. Interleukin-6 (Il6) mRNA levels were dramatically upregulated in pancreas of PGC-1α KO mice after cerulein-induced pancreatitis in comparison with wild-type mice with pancreatitis. Edema and the inflammatory infiltrate were more intense in pancreas from PGC-1α KO mice than in wild-type mice. The lack of PGC-1α markedly enhanced nuclear translocation of phospho-p65 and recruitment of p65 to Il6 promoter. PGC-1α bound phospho-p65 in pancreas during pancreatitis in wild-type mice. Glutathione depletion in cerulein-induced pancreatitis was more severe in KO mice than in wild-type mice. PGC-1α KO mice with pancreatitis, but not wild-type mice, exhibited increased myeloperoxidase activity in the lungs, together with alveolar wall thickening and collapse, which were abrogated by blockade of the IL-6 receptor glycoprotein 130 with LMT-28. In conclusion, obese rodents exhibit PGC-1α deficiency in the pancreas. PGC-1α acts as selective repressor of nuclear factor-κB (NF-κB) towards IL-6 in pancreas. PGC-1α deficiency markedly enhanced NF-κB-mediated upregulation of Il6 in pancreas in pancreatitis, leading to a severe inflammatory response.This work was supported by grants SAF2009‐09500 and SAF2015–71208‐R with FEDER funds from the Spanish Ministry of Economy and Competitiveness to JS and by grants SAF2015‐63904‐R with FEDER funds from the Spanish Ministry of Economy and Competitiveness and EC MSCA‐ITN‐2016‐721236 to MM. IF was recipient of a fellowship from ‘Programa de Pós‐Doutorado no Exterior (PDE)’ that belongs to the ‘Conselho Nacional de Desenvolvimento Científico e Tecnológico’ (CNPq).Peer Reviewe

Age-dependent regulation of antioxidant genes by p38α MAPK in the liver

p38α is a redox sensitive MAPK activated by pro-inflammatory cytokines and environmental, genotoxic and endoplasmic reticulum stresses. The aim of this work was to assess whether p38α controls the antioxidant defense in the liver, and if so, to elucidate the mechanism(s) involved and the age-related changes. For this purpose, we used liver-specific p38α-deficient mice at two different ages: young-mice (4 months-old) and old-mice (24 months-old). The liver of young p38α knock-out mice exhibited a decrease in GSH levels and an increase in GSSG/GSH ratio and malondialdehyde levels. However, old mice deficient in p38α had higher hepatic GSH levels and lower GSSG/GSH ratio than young p38α knock-out mice. Liver-specific p38α deficiency triggered a dramatic down-regulation of the mRNAs of the key antioxidant enzymes glutamate cysteine ligase, superoxide dismutase 1, superoxide dismutase 2, and catalase in young mice, which seems mediated by the lack of p65 recruitment to their promoters. Nrf-2 nuclear levels did not change significantly in the liver of young mice upon p38α deficiency, but nuclear levels of phospho-p65 and PGC-1α decreased in these mice. p38α-dependent activation of NF-κB seems to occur through classical IκB Kinase and via ribosomal S6 kinase1 and AKT in young mice. However, unexpectedly the long-term deficiency in p38α triggers a compensatory up-regulation of antioxidant enzymes via NF-κB activation and recruitment of p65 to their promoters. In conclusion, p38α MAPK maintains the expression of antioxidant genes in liver of young animals via NF-κΒ under basal conditions, whereas its long-term deficiency triggers compensatory up-regulation of antioxidant enzymes through NF-κΒ. Keywords: Nuclear factor ƙB, Glutathione, Glutamate cysteine ligase, Superoxide dismutase 1, Superoxide dismutase 2, And catalas

Age-dependent regulation of antioxidant genes by p38α MAPK in the liver

p38α is a redox sensitive MAPK activated by pro-inflammatory cytokines and environmental, genotoxic and endoplasmic reticulum stresses. The aim of this work was to assess whether p38α controls the antioxidant defense in the liver, and if so, to elucidate the mechanism(s) involved and the age-related changes. For this purpose, we used liver-specific p38α-deficient mice at two different ages: young-mice (4 months-old) and old-mice (24 months-old). The liver of young p38α knock-out mice exhibited a decrease in GSH levels and an increase in GSSG/GSH ratio and malondialdehyde levels. However, old mice deficient in p38α had higher hepatic GSH levels and lower GSSG/GSH ratio than young p38α knock-out mice. Liver-specific p38α deficiency triggered a dramatic down-regulation of the mRNAs of the key antioxidant enzymes glutamate cysteine ligase, superoxide dismutase 1, superoxide dismutase 2, and catalase in young mice, which seems mediated by the lack of p65 recruitment to their promoters. Nrf-2 nuclear levels did not change significantly in the liver of young mice upon p38α deficiency, but nuclear levels of phospho-p65 and PGC-1α decreased in these mice. p38α-dependent activation of NF-κB seems to occur through classical IκB Kinase and via ribosomal S6 kinase1 and AKT in young mice. However, unexpectedly the long-term deficiency in p38α triggers a compensatory up-regulation of antioxidant enzymes via NF-κB activation and recruitment of p65 to their promoters. In conclusion, p38α MAPK maintains the expression of antioxidant genes in liver of young animals via NF-κΒ under basal conditions, whereas its long-term deficiency triggers compensatory up-regulation of antioxidant enzymes through NF-κΒ

p38α deficiency restrains liver regeneration after partial hepatectomy triggering oxidative stress and liver injury

p38α MAPK negatively regulates the G1/S and G2/M cell cycle transitions. However, liver-specific p38α deficiency impairs cytokinesis and reduces hepatocyte proliferation during cirrhosis and aging in mice. In this work, we have studied how p38α down-regulation affects hepatocyte proliferation after partial hepatectomy, focusing on mitotic progression, cytokinesis and oxidative stress. We found that p38α deficiency triggered up-regulation of cyclins A1, B1, B2, and D1 under basal conditions and after hepatectomy. Moreover, p38α-deficient hepatocytes showed enhanced binucleation and increased levels of phospho-histone H3 but impaired phosphorylation of MNK1 after hepatectomy. The recovery of liver mass was transiently delayed in mice with p38α-deficient hepatocytes vs wild type mice. We also found that p38α deficiency caused glutathione oxidation in the liver, increased plasma aminotransferases and lactate dehydrogenase activities, and decreased plasma protein levels after hepatectomy. Interestingly, p38α silencing in isolated hepatocytes markedly decreased phospho-MNK1 levels, and silencing of either p38α or Mnk1 enhanced binucleation of hepatocytes in culture. In conclusion, p38α deficiency impairs mitotic progression in hepatocytes and restrains the recovery of liver mass after partial hepatectomy. Our results also indicate that p38α regulates cytokinesis by activating MNK1 and redox modulation.A.M.T. was recipient of a fellowship from the Ministry of Economy and Competitiveness. This work was supported by Grants SAF 2015-71208-R with FEDER funds and CSD-2007-00020 from the Spanish Ministry of Economy and Competitiveness (MINECO, Spain, http://www.mineco.gob.es/portal/site/mineco/) and GV PROMETEO II 2014-056 from Generalitat Valenciana to J.S. and R.T.V., and by Grant SAF 2015- 65267-R (MINECO/FEDER) and by Centro de Investigación Biomédica en Red de Diabetes y Enfermedades Metabólicas Asociadas (CIBERdem, ISCIII, Spain) to A.M.V.Peer reviewe

Long-Term Aspartame Administration Leads to Fibrosis, Inflammasome Activation, and Gluconeogenesis Impairment in the Liver of Mice

Background: Aspartame is an artificial sweetener used in foods and beverages worldwide. However, it is linked to oxidative stress, inflammation, and liver damage through mechanisms that are not fully elucidated yet. This work aimed to investigate the effects of long-term administration of aspartame on the oxidative and inflammatory mechanisms associated with liver fibrosis progression in mice. Methods: Mice were divided into two groups with six animals each: control and aspartame. Aspartame (80 mg/kg, via oral) or vehicle was administrated for 12 weeks. Results: Aspartame caused liver damage and elevated serum transaminase levels. Aspartame also generated liver fibrosis, as evidenced by histology analysis, and pro-fibrotic markers’ upregulation, including transforming growth factor β 1, collagen type I alpha 1, and alpha-smooth muscle actin. Furthermore, aspartame reduced nuclear factor erythroid 2-related factor 2 (Nrf2) activation and enzymatic antioxidant activity and increased lipid peroxidation, which triggered NOD-like receptor containing protein 3 (NLRP3) inflammasome activation and p53 induction. Furthermore, aspartame reduced peroxisome proliferator-activated receptor gamma coactivator 1 alpha (PGC-1α) levels, possibly through p53 activation. This PGC-1α deficiency could be responsible for the changes in lipid profile in serum, total lipid accumulation, and gluconeogenesis impairment in liver, evidenced by the gluconeogenic enzymes’ downregulation, thus causing hypoglycemia. Conclusions: This work provides new insights to understand the mechanisms related to the adverse effects of aspartame on liver tissue