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Dietary extra virgin olive oil polyphenols supplementation modulates DSS-induced chronic colitis in mice Susana Sánchez-Fidalgo, Ana Cárdeno, Marina Sánchez-Hidalgo, Marina Aparicio-Soto, Catalina Alarcón de la Lastra Journal of Nutritional Biochemistry Volume 24, Issue 7, Pages (July 2013) DOI: /j.jnutbio Copyright © 2013 Elsevier Inc. Terms and Conditions
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Fig. 1 Effect of EVOO polyphenol supplementation on DSS-induced chronic colitis. Mice received DSS for 5 days and drinking water for the next 3 weeks. Animals were fed with four different diets: SD elaborated with sunflower oil, EVOO and both supplemented with PE (SD+PE and EVOO+PE, respectively). (A) Body weight was calculated by dividing body weight on the specified day by body weight at Day 0 and expressed in percentage. (B) DAI was evaluated as average of score of clinical parameters as body weight changes, rectal bleeding and stool consistency or diarrhea. (C) Weight/length of the colon of animals treated with DSS expressed in percentage respect to control sham. Data are reported as means±S.E.M. ***P<.001 vs. control sham group; +P<.05 and +++P<.001 vs. SD-DSS group; ###P<.001 vs. SD+PE-DSS group; &P<.05, &&P<.01 and &&&P<.001 vs. EVOO DSS group. Journal of Nutritional Biochemistry , DOI: ( /j.jnutbio ) Copyright © 2013 Elsevier Inc. Terms and Conditions
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Fig. 2 Histopathology sections of proximal, middle and rectum of colonic lesions from mice fed with SD (A–C) or with EVOO+PE (D–F) and treated with 3% DSS for 5 days followed by 3 weeks of water (hematoxylin and eosin stain; original magnification ×200). (G) Histological analysis, three colonic sections of each animal were scored in a blinded fashion by a pathologist who was unaware of the experimental protocol using a colitis score as previously described by Dieleman et al. The resulting scores showed a significant reduction in histological changes in animals fed with all diets in study in proximal and middle sections. Data are reported as means±S.E.M. +++P<.001 vs. SD-DSS group. Journal of Nutritional Biochemistry , DOI: ( /j.jnutbio ) Copyright © 2013 Elsevier Inc. Terms and Conditions
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Fig. 3 Effect of EVOO and EVOO+PE on PCNA expression by immunohistochemistry method in the animal model of DSS-induced UC. Representative middle colon samples from group fed with SD (A) and group fed with EVOO+PE (B) and treated with 3% DSS for 5 days followed by 3 weeks of water. Original magnification ×200. (C) Immunohistochemical analysis for PCNA by means of the observation of three arbitrary high-power fields from colon of four animals from each diet-DSS group. The number of positive cells in 100 cells from each field was counted and the average of these values was calculated. Data are reported as means±S.E.M. +P<.05 and +++P<.001 vs. SD-DSS group; #P<.05 vs. SD+PE-DSS group; &&P<.01 vs. EVOO DSS group. Journal of Nutritional Biochemistry , DOI: ( /j.jnutbio ) Copyright © 2013 Elsevier Inc. Terms and Conditions
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Fig. 4 Effect of EVOO and EVOO+PE on MCP-1 and TNF-α mRNA in the animal model of DSS-induced UC. RT-PCR analyses of MCP-1 (A) and TNF-α (B) mRNA expression were evaluated. Values of MCP-1 and TNF-α mRNA relative expression in colon are shown as means±S.E.M. from samples (n=4) in triplicate. Data for each diet-DSS group samples were normalized by those obtained for each diet-sham group. β-Actin was used as a housekeeping gene. *P<.05, **P<.01 and ***P<.001 vs. control sham group; ++P<.01 and +++P<.001 vs. SD-DSS group; &P<.05 vs. EVOO DSS group. Journal of Nutritional Biochemistry , DOI: ( /j.jnutbio ) Copyright © 2013 Elsevier Inc. Terms and Conditions
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Fig. 5 Effect of EVOO and EVOO+PE diet on the expression of iNOS and COX-2 in the colon tissue after 3% of DSS for 5 days followed by 3 weeks of water. Densitometry was performed following normalization to the control (β-actin housekeeping gene). Data are expressed as the means±S.E.M. ***P<.001 vs. control sham group; ++P<.01 and +++P<.001 vs. SD-DSS group; ###P<.001 vs. SD+PE-DSS group; &&&P<.001 vs. EVOO DSS group. Journal of Nutritional Biochemistry , DOI: ( /j.jnutbio ) Copyright © 2013 Elsevier Inc. Terms and Conditions
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Fig. 6 Effect of EVOO and EVOO+PE diet on p-p38, p-JUNK and p-ERK1/2 activation in the colon tissue after 3% of DSS for 5 days followed by 3 weeks of water. Densitometry was performed following normalization to the control (p38, JUNK and ERK1/2 housekeeping genes, respectively). Data are expressed as the means±S.E.M. *P<.05, **P<.01 and ***P<.001 vs. control sham group; +P<.05, ++P<.01 and +++P<.001 vs. SD-DSS group; ##P<.01 vs. SD+PE-DSS group; &&&P<.001 vs. EVOO DSS group. Journal of Nutritional Biochemistry , DOI: ( /j.jnutbio ) Copyright © 2013 Elsevier Inc. Terms and Conditions
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Fig. 7 Effect of EVOO and EVOO+PE diet on IκBα degradation and PPARγ in colonic mucosa after 3% of DSS for 5 days followed by 3 weeks of water. Densitometry was performed following normalization to the control (β-actin housekeeping gene). Data are expressed as the means±S.E.M. *P<.05, **P<.01 and ***P<.001 vs. control sham group; +P<.05 and +++P<.001 vs. SD-DSS group; ##P<.01 and ###P<.001 vs. SD+PE-DSS group; &P<.05 and &&&P<.001 vs. EVOO DSS group. Journal of Nutritional Biochemistry , DOI: ( /j.jnutbio ) Copyright © 2013 Elsevier Inc. Terms and Conditions
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