Análise de viabilidade celular em linhagem RPE - 1 para teste de toxicidade do veneno de Bothrops jararaca (Wied - Neuwied, 1824)

Abstract

Snake envenomation is classified by the WHO as a neglected tropical disease. Since 2019, due to the 138,000 deaths/year, it has become a target of the organization's plan to reduce deaths and disability. In Brazil, Bothrops jararaca is responsible for 86.8% of accidents, being the subject of many studies to better understand the mechanisms of action of its venom. Toxicity testing commonly involves the use of animal models. Created in 1959, the 3 R’s principle aims to reduce, refine and replace the use of animals with alternative methods with equivalent results. Based on this premise, the objective of this work was to evaluate the toxicity of B. jararaca venom in cell culture as an alternative or complement to toxicity tests performed on animals, aiming at a greater alignment with the 3 R’s principle. RPE-1 cells (15.000 cells/well) were treated with B. jararaca venom between 0.1 μg/mL to 200 μg/mL for 24 h, cell viability and IC50 were calculated by MTT colorimetric assay. Five initial trials established optimal venom concentrations between 10 μg/mL and 200 μg/mL. The results indicated a dose-dependent decrease in cell viability, with a significant drop from 50 μg/mL, presenting viability of 70% and decreasing up to 7% at 200 μg/m. The IC50 calculation resulted in 75.20 μg/mL In conclusion, the work showed that the use of cell culture to analyze cell viability through the MTT test represents an effective method for obtaining data on the cytotoxicity of B. jararaca venom, contributing to the development of alternative methods to the use of animals, according to the principle of the 3 R’s.