Avaliação do efeito anti-inflamatório da Lectina Manose Ligante obtida das sementes de Machaerium acutifolium Vogel

Abstract

The inflammatory reaction consists of a response of the organism in the presence of an infection or injury, mediated by chemical substances that recognize the harmful stimulus as an end of destruction. Vegetable agents have been studied because they have greater safety, better efficacy, and a more economical way to treat inflammation, therefore, studies with lectins, a group of proteins of non-immune origin that have a capacity to selectively bind to carbohydrates and reversible, must be important in the medical, chemical and biological fields, in view of the importance of carbohydrates in biological processes. Therefore, the aim of this study was to evaluate the anti-inflammatory effect of lectin obtained from Machaerium acutifolium (MaL) seeds in mouse models with acute and chronic inflammation and models of macrophages stimulated by LPS. The protein fraction obtained was used for purification in affinity chromatography, followed by ion-exchange chromatography. To analyze the purity of the active fraction, they were subjected to electrophoresis under denaturing conditions. In order to assess the anti-inflammatory activity, in vivo tests were performed using male Swiss mice (20-30), for an acute phase, the formalin test was performed. Paw edema and peritonitis models were performed, along with the determination of leukocyte migration. Chronic inflammation was assessed by granuloma tests. In vitro assays were performed, using RAW 264.7 macrophages to check the cytotoxicity of MaL, as well as quantification of cytokines by plating cells treated with MaL (0.02 mg/kg, 1 mg/kg, and 5 mg/kg). A real-time quantitative polymerase chain reaction (RT-qPCR) was performed using the QuantStudioFlex6 K (Applied Biosystems) instrument. MaL significantly reduced the time to lick the mice's paws in the formalin test, both in the neurogenic phase (42.64%, 68.40%, and 59.35%) and in the inflammatory phase (50.64%, 67, 84%, and 65.60%). In carrageenan-induced paw edema, MaL inhibited the time course of carrageenan-induced edema (300 μg / paw; sc) at all doses tested (0.02 mg/kg, 1 mg/kg, and 5 mg/kg) and in the model induced by dextran, it presents a reduction in the edema percentages (41.13%). In the test of acute inflammation induced by carrageenan in the peritoneum, MaL (1.0 mg/kg) reduced 68% of the leukocyte count. MaL (1.0 mg/kg) reduces carrageenan-induced albumin leakage by 34%. In the cytotoxicity assay, MaL did not reduce cell viability in RAW 264.7 cells to a concentration of 31.25 μg / mL. In addition, one can observe the activity of MaL in the modulation of cytokines (TNF-α, IL-10, TLR2 / 4, and iNOS). Thus, MaL has anti-inflammatory activity in vivo and in vitro.