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Doctoral Thesis
DOI
https://doi.org/10.11606/T.25.2013.tde-04062013-091711
Document
Author
Full name
Josilene Luciene Duarte
Institute/School/College
Knowledge Area
Date of Defense
Published
Bauru, 2013
Supervisor
Committee
Buzalaf, Marilia Afonso Rabelo (President)
Alvarenga, Katia de Freitas
Martins, Carlos Henrique Ferreira
Olympio, Kelly Polido Kaneshiro
Saes, Sandra de Oliveira
Title in Portuguese
Potencial evocado auditivo cortical, análise bioquímica e morfológica em cérebro de ratos submetidos a tratamento subcrônico com chumbo: efeito protetor da suplementação com ferro
Keywords in Portuguese
Cérebro
Chumbo
Ferro
Morfologia
Potenciais evocados auditivos
Abstract in Portuguese
O chumbo (Pb) é um metal pesado que pode ocasionar alterações nos sistemas nervoso central (SNC) e auditivo. Faltam estudos que demonstrem tais alterações clínicas, em conjunto com as alterações bioquímicas e morfológicas em cérebro de ratos expostos ao Pb. Tem sido relatado que o ferro (Fe) tem um efeito protetor na toxicidade cerebral causada pelo Pb. O objetivo deste estudo foi avaliar a atividade elétrica cerebral, analisar a concentração de Pb no sangue e tecido cerebral, e observar o aspecto morfológico do cérebro de ratos intoxicados por Pb, submetidos à suplementação com Fe ou não. Foi realizado um projeto piloto utilizando 20 ratos machos recém-desmamados (Rattus norvegicus, variedade Wistar), que foram divididos em 5 grupos, sendo um controle e 4 experimentais (n=4/grupo). Nos grupos experimentais, os animais receberam por 6 semanas acetato de chumbo em água de beber nas concentrações de 100 mg/L e 400 mg/L de Pb, e em dois destes grupos houve administração simultânea de 20 mg/Kg de FeSO4 a cada 2 dias, por gavagem gástrica, e no outro não. O grupo controle recebeu, água deionizada. Após estabelecida a relação entre o Pb administrado e a dosagem deste no sangue (10 μgPb/dL e 30 μgPb/dL para as dosagens 100 e 400 mg/L de Pb, respectivamente), o experimento foi realizado com 30 ratos recém-desmamados divididos em 6 grupos (n=5/grupo). Os cinco primeiros foram idênticos aos empregados no estudo piloto, incluindo um grupo no qual os animais receberam água deionizada e 20 mg/Kg FeSO4 por gavagem gástrica a cada 2 dias, por 6 semanas. Após o período experimental, foi realizado o Potencial Evocado Auditivo Cortical (PEAC) com a análise da latência dos componentes P1, N1 e P2. Após a eutanásia, o cérebro dos animais foi removido, submetido ao processamento histológico (hematoxilina e eosina) e analisado ao microscópio óptico. Os dados foram analisados por ANOVA/Tukey, Kruskal-Wallis/Dunn e regressão linear (p<0,05). Foi observada uma dose-resposta em relação à concentração de Pb no sangue e no cérebro. A administração de FeSO4 reduziu os níveis de Pb no sangue e no cérebro, e embora a diferença só tenha sido significativa para o sangue (grupo que recebeu 100 mg/L de Pb), houve correlação significativa entre os níveis de Pb no sangue e cérebro (r2 = 0,94; p<0,0001). A latência média dos componentes P1, N1 e P2 do PEAC foi maior para os animais do grupo 400 mg/L Pb + 20 mg/Kg FeSO4. Não foram observadas alterações morfológicas significativas no tecido cerebral dos animais dos diferentes grupos, com exceção de alguns espaços pericelulares nos animais do grupo 400 mg/L Pb + 20 mg/Kg FeSO4. Conclui-se que a ingestão de Pb sozinho não foi capaz de causar aumentou nas latências dos componentes P1, N1 e P2 do PEAC e alteração morfológica no tecido cerebral, com exceção da maior dose de Pb utilizada (400 mg/L), associada ao uso do Fe (20 mg FeSO4). Além disso, a suplementação com o Fe foi capaz de diminuir os níveis do metal no sangue e no cérebro.
Title in English
Cortical Auditory Evoked Potential, biochemical and morphological analysis in brain of rats treated with subchronic lead: protective effect of supplementation with iron
Keywords in English
Auditory evoked potentials
Cerebrum
Iron
Lead
Morphology
Abstract in English
Lead (Pb) is a heavy metal that may cause alterations in the central nervous system (CNS) and auditory system. There is lack of studies demonstrating these clinical alterations, combined to the biochemical and morphological alterations in the brain of rats exposed to Pb. It has been reported that iron (Fe) has a protective effect on the brain toxicity caused by Pb. This study evaluated the electrical brain activity, analyzed the Pb concentration in blood and brain tissue, and observed the morphological aspect of the brain in rats intoxicated by Pb, submitted or not to Fe supplementation. A pilot study was conducted on 20 recently weaned male rats (Rattus norvegicus, Wistar type) divided in 5 groups, being one control and 4 experimental (n=4/group). In the experimental groups, the animals received lead acetate in drinking water for 6 weeks at concentrations of 100 mg/L and 400 mg/L of Pb, and in two of these groups there was simultaneous administration of 20 mg/Kg of FeSO4 at every 2 days, by gastric gavage. The control group received deionized water. After establishing the relationship between the administered Pb and its concentration in blood (10 μgPb/dL and 30 μgPb/dL for concentrations of 100 and 400 mg/L of Pb, respectively), the study was conducted on 30 recently weaned rats divided in 6 groups (n=5/group). The five first groups were identical to those used in the pilot study, including one group in which the animals received deionized water and 20 mg/Kg FeSO4 by gastric gavage at every 2 days, for 6 weeks. After the study period, the Cortical Auditory Evoked Potential (CAEC) was applied for analysis of latency of the components P1, N1 and P2. The animals were killed, their brains were removed and submitted to histological processing (hematoxylin and eosin) for light microscopy analysis. Data were analyzed by the ANOVA/Tukey, Kruskal-Wallis/Dunn and linear regression tests (p<0.05). A dose- response relationship was observed in the Pb concentration in blood and brain. The administration of FeSO4 reduced the Pb levels in blood and brain, and even though the difference was only significant for blood (group receiving 100 mg/L of Pb), there was significant correlation between the Pb levels in blood and brain ((r2 = 0.94; p<0.0001). The mean latency of components P1, N1 and P2 of the CAEC was higher in animals in the group 400 mg/L Pb + 20 mg/Kg FeSO4. No significant morphological alterations were observed in the brain tissue of animals in the different groups, except for some pericellular spaces in animals in the group 400 mg/L Pb + 20 mg/Kg FeSO4. It was concluded that Pb ingestion individually was not able to increase the latencies of components P1, N1 and P2 of the CAEC and morphological alterations in the brain tissue, except for the highest Pb dose used (400 mg/L), associated to the use of Fe (20 mg FeSO4). Additionally, the Fe supplementation was able to reduce the levels of the metal in the blood and brain.
 
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Publishing Date
2013-06-05
 
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