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P090. Differential effects of sulphasalazine, azathioprine and prednisolone on the MAP-kinases gene expression in the TNBS-induced intestinal inflammation

A. Quaglio1, A. Castilho1, L. Di Stasi1, 1Universidade Estadual Paulista (UNESP), Pharmacology, Botucatu, Brazil

Background

Inflammatory bowel disease (IBD) comprises two distinct diseases: Crohn's disease (CD) and ulcerative colitis (UC). Aetiology of this disease has not been fully elucidated but is currently presumed to result from a complex interplay among genetic, environmental, microbial and immune factors. Animal models of intestinal inflammation have given researchers a better understanding of the mechanisms involved in the pathogenesis of inflammatory bowel disease. Based on this, the aim of this study was to investigate the profile of 13 genes from MAPK family in intestinal inflammatory process induced by trinitrobenzenesulphonic acid (TNBS) and to evaluate the effects of sulphasalazine, prednisolone and azathioprine on these gene expressions.

Methods

Intestinal inflammation was induced in rat by intracolonic instillation of TNBS. Rats orally received 50 mg/kg of sulphasalazine, 2 mg/kg of prednisolone or 2 mg/kg of azathioprine at 96, 72, 48, 24 and 2 hours before colitis induction and were killed 48h after that. Colonic segments were obtained for macroscopic (colon weight and length and macroscopic damage score) and biochemical evaluation of myeloperoxidase (MPO) and alkaline phosphatase (AP) activities and glutathione (GSH) level. Gene expression analysis of MAPK1, MAPK3, MAPK4, MAPK6, MAPK7, MAPK8, MAPK9, MAPK10, MAPK11, MAPK12, MAPK13, MAPK14 and MAPK15 were performed on 100 mg of colon tissue using qRT-PCR.

Results

MAPK1, MAPK3 and MAPK9 gene expression were increased in colitic animals when compared to healthy animals indicating the involvement of these genes in intestinal inflammatory process induced by TNBS. The three drugs used were capable of reduce this expression. On the other hand, MAPK6 gene expression appeared diminished in colitic animals and treatment with sulphasalazine or azathioprine was capable to prevent it depletion. Interestingly, treatment of healthy animals with prednisolone also diminished this expression. MAPK4, MAPK7, MAPK8, MAPK10, MAPK11, MAPK12, MAPK13, MAPK14 and MAPK15 gene expression was not altered in colitic animals but surprisingly, gene expression of MAPK7 and MAPK11 was enhanced when healthy animals were treated with azathioprine.

Conclusion

MAPK1, MAPK3, MAPK6 and MAPK9 participate on the intestinal inflammatory process induced by TNBS in rats and lead to new molecular markers or therapeutic targets. Furthermore, sulphasalazine, prednisolone and azathioprine produced differential pharmacological effects on these genes expression.

Support: FAPESP and CAPES.