Impacto toxicogenômico do bisfenol A em diferentes modelos experimentais e potencial efeito reversor de um multisuplemento alimentar

Abstract

Introduction: Bisphenol A (BPA) is a synthetic monomer widely used in the plastics industry. Considered an environmental pollutant, it acts as an endocrine disruptor. Its free forms in the body are associated with the development of chronic non-communicable diseases (NCDs). However, results related to the biological mechanisms through which BPA acts are still conflicting. It is possible that BPA causes oxidative and/or inflammatory imbalance regardless of the type of organism or cell. Therefore, to mitigate these effects, many studies focus on the properties of fruits and natural compounds, associated or isolated. Objective: to evaluate the toxicogenomic impact of BPA on oxidative, genotoxic, and inflammatory markers using an in vivo model of Californian red earthworms (Eisenia fetida) and an in vitro culture model of human retinal pigment epithelium cells (ARPE-19) and characterize the reversing potential of a multi-supplement based on Guarana (Gua), Selenium (Se) and L-Carnitine (LC) (GSC®). Methodology: The work is divided into two articles. In article 1, the impact of BPA on geno-oxy-inflammatory markers was evaluated in vivo: a) the effect of acute exposure (24 h and 72 h) of adult earthworms to different concentrations of BPA (0.1, 0.3, 1 , 10 and 30 μM); b) BPA cytotoxicity and modulation of coelomocyte proliferation were determined by flow cytometry; c) The following oxidative markers were quantified by spectrophotometry: superoxide anion (O2-), nitric oxide (NO), lipoperoxidation (TBARS) and protein carbonylation (PCarb); d) The impact of BPA on coelomocyte DNA damage was determined via micronucleus (MN) assay; e) BPA changes in the immune efficiency of earthworms were tested through the coelomocyte exposure assay to dead yeast in which the formation of extracellular DNA networks and brown bodies was evaluated through cytohistological analyses; f) Genomic effect of BPA in modulating the expression of the tool-like receptor (eaTLR) and lumbricin 1 genes, via quantitative reverse transcriptase polymerase chain reaction (qRT-PCR). In article 2, cells of the ARPE-19 lineage were exposed to BPA (0.001, 0.01, 0.1, 1, 3 and 10 μM) to determine the minimum concentration to induce cytogenotoxicity. b) ARPE-19 cells were then concomitantly exposed to the selected concentration of BPA, GSC and their isolated components (Gua, 1.07 mg/mL; Se, 0.178 μg/mL; and LC, 1.43 mg/mL); c) Flow cytometry, biochemical assays, qRT-PCR, genotoxicity, apoptosis and cell proliferation were evaluated. Results: The execution of protocol 1 showed that low concentrations of BPA induced an increase in the rate of coelomocyte proliferation, imbalance in the O2-/ON ratio, high frequency of MN and apoptosis. BPA also induced increased expression of the AMP1 gene, but lower efficiency in capturing dead yeast. Already, the work carried out by protocol 2 results suggest that 10 μM of BPA is sufficient to induce cytogenotoxic changes. BPA did not change the expression ratio of Bcl-2/BAX, but induced overexpression of Casp3 and Casp8, suggesting that apoptosis was mainly induced by the extrinsic pathway. GSC partially reversed the changes triggered by BPA in ARPE-19 cells. However, Se had unexpected negative effects on ARPE-19 cells. Conclusion: These results suggested that BPA induces relevant changes, regardless of the model studied, which may be associated with an increased risk of NCDs. GSC® can attenuate changes in oxidative and genotoxic markers related to exposure of ARPE-19 cells, however, its antioxidant, antiapoptotic and genoprotective properties were not universally shared, since Se did not have a positive impact.