RESUMO
One of the theories related to aging is the increase in oxidative stress. Given this, the objective of the study is to evaluate the cellular mechanisms responsible for the resveratrol antioxidant effect on leukocytes from donors aged between 20 and 80 years old. For this, leukocytes from donors of three age groups (20-39, 40-59 and 60-80) were isolated. Image-iT™LIVE Green Reactive Oxygen Species (ROS) Kit was used. Reactive Nitrogen Species (RNS) analysis was performed by measuring nitric oxide and peroxynitrite. The PKA, Akt/PKB and p38-MAPK were evaluated by chemiluminescence. The statistical analysis between age and treatments were performed by Pearson correlation (*p < 0.05). It was possible to observe the antioxidant effect of resveratrol in all age groups. The correlation results show loss of resveratrol effect in decreasing ROS in leukocytes from older donors. We observed an active antioxidant effect of p38-MAPK in all ages, with resveratrol acting on it. The PKA and Akt/PKB were active in leukocytes from donors aged 20-59. In cells from donors older than 60, these pathways are silenced, and an effect is also not observed in cells treated with resveratrol. Therefore, resveratrol showed antioxidant effect in all age, although it was more pronounced in leukocytes from younger. One of resveratrol's mechanisms is due to the activation of the PKA and Akt/PKB, which were activated in younger donor cells.
Assuntos
Antioxidantes , Proteínas Proto-Oncogênicas c-akt , Antioxidantes/farmacologia , Resveratrol/farmacologia , Proteínas Proto-Oncogênicas c-akt/metabolismo , Proteínas Proto-Oncogênicas c-akt/farmacologia , Espécies Reativas de Oxigênio/metabolismo , Transdução de Sinais , Proteínas Quinases p38 Ativadas por Mitógeno/metabolismoRESUMO
Currently, the important role of oxidative stress in the aging process and in neurodegenerative diseases has been highlighted, suggesting the beneficial effect of antioxidants as adjuvant therapy. Resveratrol (RSV) is a polyphenolic compound used in the clinic and has been shown as an antioxidant and anti-inflammatory. Therefore, the objective was to verify neuroprotective and modulating effects of RSV on N2-A cells, pre or post inserted into an oxidative stress environment. For this, two treatment conditions were established: pre-stimulus and post-stimulus. The analysis of AMPK and SIRT1 cell signaling pathways was performed through the chemiluminescence assay using the dorsomorphin and EX527 inhibitors, respectively. The inflammatory profile was also evaluated in these neural cells, through the levels of IL-6, TNF, and IL-10. We observed that RSV in N2-A cells has anti-inflammatory effect and antioxidant property and it mechanism is dependent on the SIRT1 signaling pathway. RSV effects occurs most markedly when cells have been pre-stimulated before inducing an oxidative stress environment. These results are important for conducting more adequate protocols in the medical and nutritional clinic.