Correlação entre ingestão de aflatoxina B1, concentração sérica e urinária de AFB1-adutos e expressão hepática de marcadores moleculares relacionados à hepatocarcinogênese em ratos [thesis]

Mauricio de Rosa Trotta
Trotta MR. Correlation between aflatoxin B 1 intake and serum and urinary concentrations of AFB 1 -adducts and hepatic expression of molecular markers related to hepatocarcinogenesis in rats [thesis]. São Paulo: Universidade de São Paulo, Faculdade de Medicina de Ribeirão Preto, 2016. 101 f. Aflatoxin B1 (AFB1) is metabolite produced by fungi of genus Aspergillus that grows naturally in food. Due to weather conditions and inadequate agricultural practices, developing countries, including
more » ... s, including Brazil, have high possibility of exposure to AFB1contamined food. Chronic exposure to this mycotoxin may result in the emergence of hepatocellular carcinoma and explain the incidence of this tumor in the absence of factors such viral hepatitis and cirrhosis. After oral ingestion, AFB1 is biotransformed to its genotoxic form that binds to DNA in liver cells. This leads mutations that may be considered promoters of hepatocarcinogenesis. Following this process, there is the formation of new adducts of aflatoxins that can bind to plasma proteins or are excreted in the urine, respectively, AFB1-lysine and AFB1-N7-guanine. These compounds can be detected and work as biomarkers of exposure and toxicity of AFB1. AFB1 was administered in Wistar rats enterally, via gavage, for 90 days, and this form of exposure is closest which humans are susceptible. The animals were separated into four groups: control group (without AFB1), AFB50 (50 ppb), AFB100 (100 ppb) and AFB200 (200 ppb), in which concentration of AFB1 in part per billion (ppb) per kilogram of diet consumed by animals. It were performed liver biochemistry plasma aspartate aminotransferase (AST) and alanine aminotransferase (ALT) assessments; changes in hepatic expression of genes and proteins related to hepatocarcinogenesis (Cyclin D1, p53, β-catenin, Prohibitin, p27 Kip1 e Glutatione-S-Transferase-p1-GST-P) by immunohistochemical and real-time PCR techniques. The levels of AFB1 adducts of serum and urine were performed. The results showed increase in AST and ALT levels in all groups receiving AFB1. In group AFB200 and, lesser extent in AFB100, emerged several altered hepatocyte foci positively marked with GST-P, which are well determined preneoplastic lesions and deemed endpoints in experimental hepatocarcinogenesis assays. Analysis of liver proteins indicated that damage from AFB1 in groups AFB200 and AFB100 showed overexpression of cyclin D1, p53, β-catenin, prohibitin, indicating their participation in ways that favor the hepatocarcinogenesis. Additionally, there was a decrease in gene expression of the p27 gene, which also indicates a favorable condition for neoplastic progression to hepatocellular carcinoma. Quantification of adducts levels in serum and urine showed that the formation of these compounds was dose-dependent with different concentrations of AFB1 employed. In addition, there was a correlation between the formation of adducts with the protein expression of Cyclin D, p53, β-catenin and Rb. Thus, it was possible experimentally to point out the key proteins involved in hepatocarcinogenesis and indicate that aflatoxin adducts in serum and urine can be useful biomarkers to measure exposure and damage caused by subchronic ingestion of AFB1.
doi:10.11606/t.17.2017.tde-06012017-145306 fatcat:yerl3m5kd5beboz5q3e22xq6t4