Avaliação do potencial anti-inflamatório agudo e crônico e imunossupressor do extrato etanólico de Pterodon polygalaeflorus através de estudos in vivo e in vitro

Abstract

The genus Pterodon belongs to the family Papilonaceas and includes five native species of Brazil: P. pubescens Benth., P. emarginatus Vog., P. apparicioi Pedersoli and P. abruptus Benth., being the object of this study the species P. polygalaeflorus Benth . Its fruits are freely traded in markets and medicinal plants used in folk medicine due to anti-rheumatic, analgesic, antiinflammatory, among other effects associated with these fruits. The main use is related to the popular anti-arthritic effect that seems to be found in oily fraction of the fruit. The aim of this study was to evaluate the ethanol extract of Pterodon polygalaeflorus (EEPpg) for their potential anti-inflammatory chronic through the model of collagen-induced arthritis (CIA) and its effect on lymphocytes in vitro, as well as the MAC-cell expansion 1 + induced by Freund's complete adjuvant (CFA). The chemical characterization of EEPpg was performed by thin layer chromatography (TLC), high-performance liquid chromatography (HPLC) and gas chromatography coupled with mass spectrometry (GC-MS), through which a range of compounds, including terpenoids polarity varied and flavonoids, were observed. In model of CIA, EEPpg significantly reduced parameters associated with the development and progression of disease and disease severity, inhibiting up to 99% its development and leading to the absence of obvious clinical signs after treatment with lower doses of the extract (0,01 mg/kg and 0.001 mg/kg). Treatment with EEPpg also reduced histopathological features typical of arthritic joints that are also observed in rheumatoid arthritis. The EEPpg significantly reduced the weight of the lymph nodes of mice as well as the absolute number of segmented, monocytes and lymphocytes in the blood. In vitro, EEPpg showed an anti-proliferative activity of splenocytes stimulated with concanavalin A (Con A) or lipopolysaccharide (LPS) analyzed by testing to reduce the tetrazolium salt MTT, supported by its effect on the cell cycle of lymphocytes stimulated with Con A, where the EEPpg at concentrations of 5, 10 and 20 mg/mL significantly reduced in concentration-dependent manner, the number of cells in phases S + G2 / M and increased G0/G1 phase of cell cycle. The anti-proliferative EEPpg seems also to be associated with increased apoptosis of lymphocytes after stimulation with Con A, with a statistically significant increase in the percentage of dead cells by apoptosis at higher concentrations. The EEPpg inhibited the expansion of Mac-1 + cells induced by AFC in the spleen but not in the peritoneum. This suggests an inhibitory effect on cell migration by EEPpg into arthritic joints. These results help to validate the popular use of P. polygalaeflorus against diseases related to inflammatory and immune disorders, especially rheumatoid arthritis, never shown before.