Caracterização química e efeito preventivo do extrato hidroetanólico liofilizado do cladódio de Cereus jamacaru P.DC. (Mandacaru) em modelo experimental de doença inflamatória intestinal

Abstract

Inflammatory bowel diseases (IBD) are chronic inflammatory pathologies associated with genetic changes and environmental factors. The drugs available for treatment can cause adverse effects, showing the need to search for new therapeutic strategies. Foods rich in bioactive compounds with anti-inflammatory activity stand out, such as Cereus jamacaru P. DC. This research aimed to evaluate the physicochemical, phytochemical and anti-inflammatory effect of the Lyophilized Hydroethanolic Extract from Cereus jamacaru P.DC cladode. (mandacaru) in an experimental model of DNBS-induced acute inflammatory bowel disease. The mandacaru cladodes were collected in the city of Jucurutu / RN, the thorns removed, cut, dried and ground, undergoing maceration with ethanol and water, obtaining the Lyophilized Hydroethanolic Extract of Cereus jamacaru (EHCJ). The EHCJ phytochemistry was performed by liquid chromatography coupled to a mass spectrometer to assess the presence of phenolic compounds. In the study of intestinal anti-inflammatory activity, Rattus norvegicus (Wistar) (n = 8 / group), weight 230 ± 20 g, randomly distributed in the groups: normal control, DNBS, sulfasalazine 250 mg / kg and EHCJ (50, 100 and 200) were used mg/kg/day). Disease activity index (IAD), preliminary signs of acute toxicity, macroscopic, microscopic, volatile compounds and fatty acids do not present fecal material were evaluated. Furthermore, inflammatory markers such as myeloperoxidase (MPO) dosage, an expression of the mitogen-activated protein kinase (MAPK) gene, the metalloproteinase 9 protein (MMP-9) and the kappa B factor gene and protein (NF-κB p65) ); cytokine dosages (TNF-α, IL-10 and IL-1β); evaluation of oxidative stress by measuring malondialdehyde (MDA) and reduced glutathione (GSH); integrity of the intestinal barrier through the expression of the occludin gene (OCL), expression of the mucin 2 (MUC-2) and occlusion zone (ZO-1) proteins, all in colonic tissue. In phytochemistry, compounds derived from benzoic acid, saccharide, Kampferol-3-O-dirhamnosyl-hexoside, Isorhamnetin-3-O-dirhamnosyl-glycoside, Kaempferol-3-O-rutinoside, Isorhamnetin-3-Orutinoside and o Oxo-dihydroxy-octadecenoic acid. Through the GC-MS of the fecal materials, eighteen compounds were identified, being them fatty acids, hydrocarbons, sterols and phenol. In the colitis model, EHCJ at all doses tested was able to cause chemoprevention observed by means of the IAD (p < 0.05). Furthermore, it protected against DNBS-induced colonic damage with a lower macroscopic damage score (p < 0.05). EHCJ promoted lower MPO activity (p < 0.05), lower MDA activity (p < 0.01) and higher levels of GSH (p < 0.05) for all doses tested. EHCJ also caused lower tissue levels of the cytokines TNF-α (p <0.0001) and IL-1β (p < 0.05) and higher levels of IL-10 at the dose of 200 mg / kg (p < 0.01) . The preventive effect was also reinforced by the higher expression of the OCL gene and lower expression of the MAPK pathway markers (p < 0.01) and NF-κB p65 (p < 0.0001) in the colon. Histopathological analyzes indistinct than EHCJ showed lower infiltrate density (p <0.001) and immunohistochemistry showed higher immunoreactivity for MUC-2 (p <0.01) and ZO-1 (p <0.05) and lower for the NF-κB proteins p65 (p<0.05) and MMP-9 (p<0.05). Thus, preclinical results indicate that EHCJ is beneficial in the chemoprevention of colitis and may, in the future, become an alternative in the treatment of human IBD.