Avaliação do efeito anti-inflamatório e anti-proliferativo de Condroitim sulfato e/ou Glucosamina em células RAW 264.7, IEC-6 e Caco-2

Abstract

Chondroitin Sulfate (CS) and Glucosamine (GlcN) are currently indicated for the treatment of inflammatory diseases, such as osteoarthritis, by reducing the action of nuclear transcription factor NF-kB, metalloproteinase activity (MMP), interleukins, nitric oxide synthase inducible (iNOS) among other inflammatory mediators. The objective of the present study was to investigate the effects of CS, GlcN,and CS/GlcN on the model of acute inflammation induced by lipopolysaccharide (LPS) in cell lines RAW 267.4, IEC-6 and Caco-2, as well as, to understand the molecular mechanism of this effect on macrophages RAW 264.7, IEC-6 and Caco-2. CS and GlcN were characterized by agarose gel electrophoresis, polyacrylamide gel for molecular weight, fluorophore-assisted carbohydrate electrophoresis (FACE), and 13C NMR. The anti-inflammatory activity of CS and GlcN associated or not was evaluated by the production of nitric oxide (NO), metalloproteinases activity (MMP-9 and -2), TNF-a, IL-6 and IL-1b by ELISA. In addition, translocation of NF-κB by confocal immunofluorescence microscopy, and expression of IL-1b and iNOS. The analyzes showed that the CS used in the tests is characterized as a chondroitin 4sulfate. CS, GlcN and CS/GlcN were able to reduce iNOS expression and NO production. In addition to reducing levels of TNF-a in RAW 264.7 macrophages. Already CS/GlcN and GlcN reduced the activity of MMP-9, and the production of IL1b, maintaining a higher pro-IL-1b expression. Cs, GlcN and CS/GlcN also decreased the nuclear translocationt of the NF-kB. Thus, these results indicate that CS and GlcN associated or not, present an anti-inflammatory activity against RAW 264.7 macrophages. With regard to the response of the compounds studied to IEC-6 and Caco-2 cells, it was not possible to evaluate in the available conditions, once, that we can not stimulate an inflammatory process in them.