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Effects of symbiotic and indole 3-carbinol on the gene expression involved in colon carcinogenesis in rats fed a diet containing heme

Abstract

The present study aims to evaluate the preventive effect of administration of synbiotic (prebiotic inulin associated with the probiotic Bifidobacterium bifidus) and indole-3-carbinol on the chemical process of colon carcinogenesis induced by 1,2-dimethylhydrazine (DMH) in male Wistar rats fed a diet containing heme. The animals will be allocated into seven groups with 10 to 22 animals each, animals in groups 1 to 6 and 7 will receive four subcutaneous injections of DMH (40 mg / kg) and EDTA solution (DMH vehicle) in the initial two weeks the experiment, respectively. Groups 1 and 7 will receive basal diet until the end of the experiment, groups 2-6 respectively receive basal diet containing heme (0.32 g / kg diet), basal diet containing heme (0.32 g / kg diet) + symbiotic (inulin + Bifidobacterium bifidum 2.5 x 1010 CFU per kg diet), basal diet containing heme (0.32 g / kg diet) + indole-3-carbinol (2g/kg diet), basal diet + symbiotic (inulin + Bifidobacterium bifidum 2.5 x 1010 CFU of feed) and basal diet + Indo-3-carbinol (2g/kg feed). The animals will be euthanized on the 12th and 22nd weeks of the experiment. The colon (medial and distal) will be removed and immersed in fixative (24 h) and stored in 70% alcohol. The colon will be stained with methylene blue 2% for the detection of stereoscopic ACF (incidence and multiplicity). After analysis of ACF, the colon will be processed to obtain the histological classification of dysplasia of the FCA and immunohistochemical detection of the expression of B-catenin (²-catenin positive ACF), Ki-67 and cleaved caspase 3. Tumor samples collected at the end of the 22nd week of the trial will be frozen in liquid nitrogen and stored at -80 °C Biofreeze. The gene expression pathways that regulate cell proliferation and apoptosis in the tumor samples will be analyzed by the technique Taqman ® Low Density Array (TLDA). Tumor samples are immersed in fixative (24 h) and stored in 70% alcohol and subsequently processed to obtain histological and immunohistochemical expression for B-catenin (B-catenin positive ACF), Ki-67 and cleaved caspase-3 and other markers selected after analysis of gene expression. (AU)

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Scientific publications
(References retrieved automatically from Web of Science and SciELO through information on FAPESP grants and their corresponding numbers as mentioned in the publications by the authors)
DE MOURA, NELCI A.; CAETANO, BRUNNO F. R.; BIDINOTTO, LUCAS T.; RODRIGUES, MARIA A. M.; BARBISAN, LUIS F.. Dietary hemin promotes colonic preneoplastic lesions and DNA damage but not tumor development in a medium-term model of colon carcinogenesis in rats. MUTATION RESEARCH-GENETIC TOXICOLOGY AND ENVIRONMENTAL MUTAGENESIS, v. 846, . (11/23699-4, 12/12631-2, 13/08033-5)
DE MOURA, NELCI A.; CAETANO, BRUNNO F. R.; DE MORAES, LEONARDO N.; CARVALHO, ROBSON F.; RODRIGUES, MARIA A. M.; BARBISAN, LUIS F.. Enhancement of colon carcinogenesis by the combination of indole-3 carbinol and synbiotics in hemin-fed rats. Food and Chemical Toxicology, v. 112, p. 11-18, . (13/08033-5, 12/12631-2, 11/23699-4)
DE MOURA, N. A.; CAETANO, B. F. R.; BIDINOTTO, L. T.; RODRIGUES, M. A. M.; BARBISAN, L. F.. Synbiotic supplementation attenuates the promoting effect of indole-3-carbinol on colon tumorigenesis. BENEFICIAL MICROBES, v. 12, n. 5, p. 493-501, . (13/08033-5, 12/12631-2, 11/23699-4)

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