Development of TNBS-induced colitis: animal model to test new pharmacological approaches

V. Mateus; P. Faísca; H. Mota-Filipe; Bruno M. Sepodes; R. Pinto

V. Mateus Escola Superior de Tecnologia da Saúde de Lisboa - IPL - Pharmacology and Translational Research Group, iMED – Faculdade de Farmácia. Universidade de Lisboa. Portugal
P. Faísca Departamento de Anatomia Patologica, Faculdade de Medicina Veterinária - ULHT. Portugal
H. Mota-Filipe Pharmacology and Translational Research Group, iMED – Faculdade de Farmácia. Universidade de Lisboa. Portugal
Bruno M. Sepodes Pharmacology and Translational Research Group, iMED – Faculdade de Farmácia. Universidade de Lisboa. Portugal
R. Pinto Pharmacology and Translational Research Group, iMED – Faculdade de Farmácia. Universidade de Lisboa. Portugal

Keywords: IBD, TNBS-induced colitis, Inflammation, Metabolic pathways, Pharmacological targets

Abstract

IBD is a gastro-intestinal disorder marked with chronic inflammation of intestinal epithelium, dam- aging mucosal tissue and manifests into several intestinal and extra-intestinal symptoms. Currently used medical therapy is able to induce and maintain the patient in remission, however no modifies or reverses the underlying pathogenic mechanism. The research of other medical approaches is crucial to the treatment of IBD and, for this, it ́s important to use animal models to mimic the characteristics of disease in real life. The aim of the study is to develop an animal model of TNBS-induced colitis to test new pharmacological approaches. TNBS was instilled intracolonic single dose as described by Morris et al. It was administered 2,5% TNBS in 50% ethanol through a catheter carefully inserted into the colon. Mice were kept in a Tredelenburg position to avoid reflux. On day 4 and 7, the animals were sacrificed by cervical dislocation. The induction was confirmed based on clinical symptoms/signs, ALP determination and histopathological analysis. At day 4, TNBS group presented a decreased body weight and an alteration of intestinal motility characterized by diarrhea, severe edema of the anus and moderate morbidity, while in the two control groups weren’t identified any alteration on the clinical symptoms/signs with an increase of the body weight. TNBS group presented the highest concentrations of ALP comparing with control groups. The histopathology analysis revealed severe necrosis of the mucosa with widespread necrosis of the intestinal glands. Severe hemorrhagic and purulent exsudates were observed in the submucosa, muscular and serosa. TNBS group presented clinical symptoms/ signs and histopathological features compatible with a correct induction of UC. The peak of manifestations became maximal at day 4 after induction. This study allows concluding that it’s possible to develop a TNBS- induced colitis 4 days after instillation.

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