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P089. Differential effects of sulphasalazine, azathioprine and prednisolone on the NF-kB, heparanase, HSP70 and HPRT 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) refers to a group of conditions characterized by inflammation in the intestinal tract that includes Crohn's disease (CD) and ulcerative colitis (UC). The aetiology of these diseases 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 NF-kB, heparanase, HSP70 and HPRT gene expression in intestinal inflammation induced by TNBS (trinitrobenzenesulphonic acid) 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. Animals were killed 48 h after colitis induction, colonic segments were obtained after laparotomy 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 NF-kB, heparanase, HSP70 and HPRT were performed on 100 mg of colon tissue using qRT-PCR.

Results

All gene expressions were increased in colitic animals when compared to healthy animals, indicating the involvement of these genes in intestinal inflammatory process induced by TNBS. Treatment of healthy animals with sulphasalazine and prednisolone did not affect gene expression, but it was observed that azathioprine enhanced HSP70 gene expression in healthy animals. On the other hand, treatment of colitic animals with sulphasalazine restored heparanase, HPRT and NF-kB gene expression, while azathioprine restored the heparanase and HPRT gene expression and prednisolone only restored gene expression of the HPRT and HSP70.

Conclusion

Gene expression of NF-kB, heparanase, HSP70 and HPRT 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 NF-kB, heparanase, HSP70 and HPRT gene expression.

Financial support: FAPESP and CAPES.