A influência da associação do laser de baixa potência e do guaraná (Paullinia cupana kunth) nos marcadores oxidativos, inflamatórios, fatores de crescimento e na proliferação de fibroblastos in vitro

Abstract

Introduction: Cutaneous aging in humans is a complex process and can be induced by intrinsic and extrinsic factors. Reactive oxygen species (ROS) are the main responsible for skin changes, especially at the level of fibroblasts. The accumulation of ROS in the organism and lack of antioxidant defenses generate oxidative stress, which is a condition that can damage the lipids and proteins of the membrane and cellular deoxyribonucleic acid (DNA). Previous investigations have shown that guaraná (Paullinia cupana var. Sorbilis Mart Kunth) may repair cell damage due to its an antioxidant and anti-inflammatory action. In addition, electrotherapeutic treatment with a low-level laser therapy (LLLT) stands out due to its anti-inflammatory, cicatrizing, and tissue biomodulating action. The association of these two therapies can be used to support traditional treatments to repair skin cell damage, which justifies the relevance of this research since there are no studies associating these therapies. Objective: To evaluate the in vitro effect of LLLT in cellular senescence markers of human fibroblasts exposed to H2O2 and in the potentiation of the cytoprotective effects associated with guaraná. Methods: This study was performed on commercial human fibroblast cell culture (line HFF-1). Cells were pre- exposed to H2O2 for 2 hours and then treated with LLLT at a wavelength of 660nm and guaraná. Cells remained in the culture for 24 or 72 hours. After the determinations of the doses, the treatments of guaraná and LLLT were performed alone and in association. After 24 or 72 hours, analyzes of the oxidative, apoptotic, and cell proliferation markers were performed according to protocols previously described in the literature. Results: The first experiment (manuscript 1) determined the dose curve of LLLT and H2O2 and the interaction of these treatments. The best dose of H2O2 for this study was 50μM and 4 J/cm2 for LLLT. At this dose, the interaction of LLLT with cells exposed to H2O2 showed that LLLT has an anti-inflammatory, anti-apoptotic, and cell proliferation-enhancing effect. The second experiment (manuscript 2) established the dose curve of guaraná and the effect of the single use and association of LLLT with guaraná in reversing the damage caused by H2O2. The results also revealed that both treatments have cytoprotective action, although this association showed more relevant results in cell proliferation due to the significant increase of cells in the S phase and FGF-1 expression in the treated fibroblasts. Conclusion: This study suggests that LLLT is able to act positively on fibroblast senescence markers and that the association of LLLT with guaraná intensified cytofunctional effects in relation to isolated treatments, especially in cell proliferation.