Impactos celulares de aditivos de aroma e sabor: abordagens in sílico in vitro com enfoque imunotoxicológico

Abstract

The increase in processed and convenience food production has made the use of food additivesan important practice in food technology. The flavorings impart aroma and flavor and are associated with the pleasure of eating and drinking. However, there is a lack of studies that measure the effects related to the use of these additives in the food industry. In this context, the main objective of this work was to evaluate the effects of synthetic flavors, γ-hexalactone and furfuryl alcohol, and the natural aroma of rosemary, through in silico and in vitro approaches. In silico tests was performed through online platforms and consisted of comparative structural analysis with structurally related substances or fragments present in databases. For the in vitro evaluation, the test concentrations were initially selected by cytotoxicity screening in lymphocyte culture or according to previous studies. The results of the computational analysis indicate mutagenic potential for γ-hexalactone and carcinogenic for both synthetic flavorings. Nevertheless, the first was not confirmed by the micronucleus frequency test, probably because the predictive analysis was based on Ames test results. On the other hand, the carcinogenic potential of γ-hexalactone and furfuryl alcohol, indicated by the PreADMET and ProTox platforms, respectively, may be related to DNA damage in the highest non-cytotoxic concentration of γ-hexalactone evaluated (5.15 μM) and from 25 μg/mL furfuryl alcohol. This damage may be due to the pro-oxidant status of γ-hexalactone, which may contribute to the initiation and propagation of reactions with biomolecules. Regarding the mechanism involved in the DNA damage of lymphocytes exposed to furfuryl alcohol, it is probably associated with the action of sulfotransferases and formation of 2-sulfoxymethylfuran which upon reacting with the DNA molecule forms adducts. The γ-hexalactone also demonstrated immunomodulatory potential in vitro by interfering with cytokine secretion/release, promoted by NK cell activation as previously studied with γ-dodecalactone. Additionally, furfuryl alcohol induced karyopyknosis which, probably and according to the computational platforms employed, is due to the down regulation of genes that act on the chromosomal architecture. Rosemary's natural aroma demonstrated concentration-dependent cytotoxic potential reducing lymphocyte subpopulations at all concentrations analyzed. Also, within 24 h of experimental intervention, 25 μg/mL rosemary caused DNA damage 35 times higher than the negative control. In addition, concentrations ranging from 1 to 50 μg/mL caused chromosomal instability and interfered with the rate of cell division. In contrast, Rosemary extract was effective in maintaining reactive species at levels of control and inhibition of lipid peroxidation, due to its antioxidant power. Thus, the results suggest that rosemary may act as a cytotoxic and/or antiproliferative agent, corroborated by the induction of genomic instability and interference in the regular process of cell division, respectively. Given the above, it is understood that synthetic flavors γ-hexalactone and furfuryl alcohol, as well as the Rosemary natural aroma may be potentially immunotoxic and or immunomodulatory.