Avaliação do mecanismo de ação anti-inflamatória do extrato metanólico e fração hexânica de Cariniana rubra Gardner ex Miers

Abstract

EVALUATION OF THE MECHANISM OF ACTION OF ANTI - INFLAMMATORY METHANOL EXTRACT AND HEXANE FRACTION OF Cariniana rubra Gardner ex miers. Silva D.N.P.B. Dissertation presented to the Coordination of the Post-Graduation Programs in Medicine of Faculty of Medicine of Federal University of Mato Grosso, as a partial requirement for the Master Degree of Health Sciences, Pharmacology Field. Advisor: Prof. Dr. Amilcar Sabino Damazo, Co -supervisor: Prof. Dr. Domingos Tabajara de Oliveira Martins. Cariniana rubra Gardner ex Miers is a tree that belongs to the family Lecythidhaeae This family is present in very different regions as areas with low flood periodically , mountainous regions with higher elevations to 1,000 feet above sea level clenched and regions with secondary vegetation ( capoeira ) formed after disturbances in natural environments . Currently has eight chemical compounds have been identified from the inner bark as phenolic compounds: β - sitosterol , stigmasterol , α and β-amirinas , arjunolic acid and sitosterol 3 - O- β - D - glucopiyanosil . Triterpenoid saponins : 28 - β - glucopyranosyl -23 - O- acetyl arjunolic acid , 3 - O- β - glucopyranosil arjunolic acid and 28 - O- [ α -L - rhamnopiranosil - ( 1 → 2 ) - β - glucopyranosil ] -23 - O- aceti0l arjunolic acid . Goal of this study was the validation of its anti-inflammatory properties, determining its mechanism of action and identification of secondary metabolites using in vivo and in vitro experimental models. To obtain the fraction of methanol extract was first subjected to liquid -liquid partition by suspending a mixture of methanol and distilled water. Then, hexane was added and after stirring the formation of the hexane fraction. Phytochemical screening revealed the existence of several secondary metabolites many of which are known anti - inflammatory action. It was possible to identify some compounds that are responsible for the anti-inflammatory action as by chromatographic and spectrometric analysis.The Hippocratic essay no animal death and organ analyzed did not change demonstrating that both EMCR as FrHCr not exhibit toxicity. In the cytotoxicity assay using cells of Chinese hamster ovary (CHO) by the method of Alamar Blue showed that both EMCR as FrHCr can be considered cytotoxic (IC50 ˂ 50 mg / ml). In an inflammation model by air pouch histological analysis demonstrated that the skin FrHCr have an EMCR and antimigratory effects of leukocytes and analysis of the washed leukocyte migration bubble demonstrated that treatment with EMCR was similar to standard drug , preventing migration PMN and MN cavity following administration of carrageenan . To understand the mechanism of action of the inflammatory process, we evaluated the release of the cytokines TNF - α, IL - 1β and IL-10 and expression of AnxA1. The AnxA1 protein expression was evaluated by immunohistochemistry data indicated that, after induction of inflammation, we observed an increase in anti-inflammatory protein AnxA1, particularly in neutrophils. The fact that EMCR induce expression of AnxA1 indicates that some phytochemical component of this plant induces the protein expression of this mediator, this being one of its possible anti-inflammatory mechanisms.