252P Granada Congress and Exhibitions Centre
6th European Congress of Pharmacology (EPHAR 2012)

 

 

The intestinal anti-inflammatory effects of the extract Amanda ® in the TNBS model of rat colitis are associated with epithelial barrier function improvement.

P Zorrilla1, L Tchombe1, MP Utrilla1, N Garrido-Mesa1, ME Rodriguez-Cabezas1, M Toral1, F Algieri1, A Rodriguez-Nogales1, L Perez2, M Olivares2, A Zarzuelo1, J Galvez1. 1CIBER-EHD University of Granada, Department of Pharmacology, Spain, 2Biosearch Life, 2Department of Immunology and Animal Sciences, Spain

 

Introduction: Different plant extracts containing polyphenols with antioxidant properties have been traditionally used in different human conditions, including inflammation. Since inflammatory bowel diseases (IBD) are associated with oxidative stress, the use of these polyphenol-enriched extracts may be of potential interest in their treatment. The aim of this study was to evaluate the intestinal anti-inflammatory properties of the extract Amanda ® , obtained from almonds, that contains at least 30 % of polyphenolic compounds, in the trinitrobenzenesulphonic acid (TNBS) model of rat colitis, a well characterized model with some resemblance to human IBD.

Material and Methods: Female Wistar rats (200 ± 10 g) were assigned to seven groups (n=8): non-colitic, control colitic (without treatment) and treated colitic groups: four treated with Amanda ® extract (10, 25, 50 and 100 mg/kg/day) (provided by Biosearch Life) and the remaining received sulphasalazine (SAZ) (100 mg/kg/day), as a positive control. Treatments started the same day of TNBS colitis induction, and rats were sacrificed one week after. Colonic damage was assessed macroscopically (score 0-10) and biochemically: myeloperoxidase activity (MPO), glutathione content (GSH), as well as IL-1β, MUC-3 and TFF-3 expressions by qPCR. Statistical analyses was carried out with Statgraphics 5.0, with statistical significance set at p<0.05 using a one-way analysis of variance (ANOVA) and post hoc least significance tests.

Results: The administration of Amanda ® extract resulted in an intestinal anti-inflammatory effect, at doses of 25 and 50 mg/kg/day, as evidenced macroscopically by a reduction in the extension of damage in comparison with control colitic rats (Table 1). Biochemically, it was observed a decrease in colonic MPO activity over 60% in the doses of 25 and 50 mg/kg and a 50% increment in glutathione content, thus improving the altered oxidative status in colitic rats. In addition, the treatment with 50 mg/kg of Amanda ® extract resulted in a three-fold decreased colonic expression of the proinflammatory cytokine IL-1β, and a three-fold higher expression of markers of intestine epithelial integrity, MUC-2, TFF-3 (Table 1).

Table 1. Effects of Amanda ® extract in TBNS rat colitis

Group (n=10) Score (0-10) MPO activity (mU/g tissue) GSH content (nmol/g tissue) IL-1 β (fold change) MUC-3 (fold change) TFF-3 (fold change)
Healthy 0.0 ± 0.0 58 ± 14 1346 ± 111 1.0 ± 0.2 1.0 ± 0.2 1.0 ± 0.2
Colitic Control 8.0 ± 0.2 2429 ± 728 282 ± 56 11.2 ± 2.1 0.3 ± 0.1 0.2 ± 0.1
Amanda ® (10 mg/kg) 8.0 ± 0.2 2141 ± 356 374 ± 138 7.4 ± 1.7 1.0 ± 0.4 0.2 ± 0.1
Amanda ® (25 mg/kg) 6.7 ± 0.3* 1034 ± 317* 708 ± 159* 7.6 ± 2.0 0.9 ± 0.3 0.5 ± 0.1
Amanda ® (50 mg/kg) 5.6 ± 0.5* 770 ± 231* 884 ± 308* 3.5 ± 1.3* 0.8 ± 0.2* 0.4 ± 0.2
Amanda ® (100 mg/kg) 6.1 ± 0.5* 2041 ± 421 601 ± 145* 8.8 ± 1.6 1.1 ± 0.2* 0.7 ± 0.2*
SAZ(100 mg/kg) 6.7 ± 0.5 1415 ± 434* 786 ± 313* 7.2 ± 0.8 0.9 ± 0.2* 0.4 ± 0.1

Data are expressed as mean ± SEM. *p<0.05 vs. Control group.

Conclusion: The Amanda ® extract showed intestinal anti-inflammatory activity in the TNBS model of rat colitis, in which the antioxidant properties ascribed to some of its components may play a role, thus resulting in a protective effect to the epithelial barrier.