Avaliação do efeito das nanopartículas poliméricas de resveratrol em células neuronais PC12 expostas à rotenona: um modelo in vitro da doença de Parkinson

Abstract

Parkinson's disease (PD) is a progressive neurodegenerative disorder resulting from the degeneration of dopaminergic neurons and the abnormal deposition of alpha-synuclein in brain cells, forming Lewy bodies. Its etiology is idiopathic, associated with environmental and genetic factors. The possible molecular mechanisms of pathogenesis are oxidative stress, mitochondrial dysfunction, and neuroinflammation. Treatment remains a challenge, as available medications, such as Levodopa, only treat motor symptoms without interfering with disease progression. Thus, new therapeutic options capable of interfering with disease progression are needed, offering increased quality of life and survival for patients. The objective of this study was to evaluate the neuroprotective effect of resveratrol polymeric nanoparticles (NPR) in an in vitro model of PD, using rotenone-induced PC12 neuronal cells (ROT). The methodology used the MTT test to define the working concentrations of the substances (ROT, RSV, NPR, and DOPA) and analyze cell viability, flow cytometry to evaluate cell death mechanisms, and optical microscopy to evaluate morphological changes resulting from exposure to rotenone and pretreatment with different substances. GraphPad Prism software was used for statistical analysis, and Shapiro-Wilk, One-Way ANOVA, and Tukey tests were applied. As a result, the working concentrations of the substances used were: ROT: 19.58 µg/mL; RSV: 12.5 µM, 6.25 µM, and 3.12 µM; NPR: 1.56 µM, 0.78 µM, and 0.39 µM; DOPA: 400 µM, 200 µM, and 100 µM. It was also demonstrated that PC12 cells had increased viability when exposed to all tested concentrations of RSV and NPR, compared to the group exposed to ROT, while treatment with DOPA did not show a significant increase (ROT: 47.6% ± 3.010%; RSV 1: 63.11% ± 2.632%; RSV 2: 69% ± 2.882%; RSV 3: 73.67% ± 2.179%; NPR 1: 73.11% ± 2.220%; NPR 2: 77.67% ± 3.528%; NPR 3: 62.78% ± 3.770%). Regarding the evaluation of the cell death pattern, treatment with RSV1 (12.5 µM) and NPR3 (NPR 3: 0.39 µM) resulted in a similar reduction in apoptotic events, indicating greater efficiency of NPRs. All DOPA concentrations were able to significantly reduce the occurrence of apoptosis. The present study revealed resveratrol as a potential cytoprotective agent, but clinical trials are needed to confirm its possible use in the treatment of PD.