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Master's Dissertation
DOI
Document
Author
Full name
Jairo Pinheiro da Silva
E-mail
Institute/School/College
Knowledge Area
Date of Defense
Published
Ribeirão Preto, 2019
Supervisor
Committee
Tirapelli, Luis Fernando (President)
Fazan, Valeria Paula Sassoli
Regalo, Simone Cecilio Hallak
Title in Portuguese
Análise da proteína CASPASE 9 e dos microRNAs miR-21, miR126 e miR-155 relacionados ao mecanismo de apoptose no cerebelo de ratos submetidos à isquemia cerebral focal associada ou não ao modelo de alcoolismo
Keywords in Portuguese
Alcoolismo
Apoptose
Cerebelo
Isquemia focal
microRNAs
Abstract in Portuguese
Introdução: A isquemia cerebral é uma desordem da função cerebral ocasionado pela supressão sanguínea no tecido cerebral sem nenhuma outra causa aparente do que a vascular. Estudos revelam os danos causados pela isquemia cerebral focal repercutem não apenas na região da lesão isquêmica, mas também em outras regiões do encéfalo, dentre elas o cerebelo. O etanol atua diminuindo o tempo de reação do corpo e a resposta reflexa, produzindo até mesmo perda de coordenação motora. Por tempos, estudos tem verificado a ação do etanol no cerebelo. Objetivos: Este trabalho tem por objetivo analisar o córtex cerebelar de ratos submetidos a um modelo experimental de isquemia cerebral focal transitória por oclusão da ACM durante 90 minutos, seguida por reperfusão de 48 horas, associado ou não a modelo de alcoolismo. Material e métodos: Foram utilizados 50 ratos Wistar adultos, subdivididos em 5 grupos experimentais: grupo controle (C): animais submetidos apenas à anestesia; grupo sham (S): animais submetidos à simulação completa do procedimento cirúrgico; grupo isquêmico (I): animais submetidos à isquemia cerebral focal por 90 minutos seguido por reperfusão de 48 horas; grupo alcoolizado (A): animais que receberam diariamente álcool etílico absoluto diluído a 20% em água durante quatro semanas; e, grupo isquêmico e alcoolizado (IA): animais submetidos ao mesmo tratamento do grupo A e que, após quatro semanas foram submetidos à isquemia cerebral focal durante 90 minutos, seguido por reperfusão de 48 horas. As amostras do cerebelo coletadas e realizado a análise de imunohistoquímica da proteína CASPASE-9 e a análise sérica por meio de PCR - RT dos miRNAS miR-21, miR-126 e o miR155. Resultados: A expressão de CASPASE 9 teve maior expressão no grupos I, A e I+A. A análise dos miRNAS, o miR-126 foi maior nos grupos A e I+A, o miR-155 foi maior nos grupos I e I+A. Conclusões: Podemos concluir que a ocorrência de apoptose no córtex cerebelar, mesmo distante do foco isquêmico e ques miRNAs 126 e 155 apresentam correlação com a apoptose celular em ratos isquêmicos e submetidos ao modelo de alcoolismo crônico
Title in English
ANALYSIS of the CASPASE 9 PROTEIN AND THE MICRORNAS MIR-21, MIR-126 AND MIR-155 RELATED TO THE MECHANISM OF APOPTOSIS IN THE CEREBELLUM OF RATS SUBMITTED TO FOCAL CEREBRAL ISCHEMIA ASSOCIATED OR NOT TO THE MODEL OF ALCOHOLISM
Keywords in English
Alcoholism
Apoptosis
Cerebellum
Focal ischemia
microRNAs
Abstract in English
INTRODUCTION: Cerebral ischemia is a disorder of brain function caused by blood suppression in brain tissue with no apparent cause other than vascular. Studies reveal the damage caused by focal cerebral ischemia to affect not only the region of the ischemic lesion but also other regions of the brain, including the cerebellum. Ethanol acts by decreasing the reaction time of the body and the reflex response, producing even loss of motor coordination. For some time, studies have verified the action of ethanol in the cerebellum. AIMS: This study aims to analyze the cerebellar cortex of rats submitted to an experimental model of transient focal cerebral ischemia by ACM occlusion for 90 minutes, followed by reperfusion of 48 hours, associated or not with the model of alcoholism. METHODS: Fifty adult Wistar rats were used, subdivided into 5 experimental groups: control group (C): animals submitted to anesthesia only; sham group (S): animals submitted to complete simulation of the surgical procedure; ischemic group (I): animals submitted to focal cerebral ischemia for 90 minutes followed by reperfusion of 48 hours; alcoholic group (A): animals that received daily absolute ethanol diluted 20% in water for four weeks; and ischemic and alcoholized group (AI): animals submitted to the same treatment as group A and after four weeks were submitted to focal cerebral ischemia for 90 minutes, followed by reperfusion of 48 hours. The cerebellum samples were collected and the immunohistochemical analysis of the CASPASE-9 protein and the serum analysis by means of RT-PCR of miRNAS miR-21, miR-126 and miR155 were performed. RESULTS: The expression of CASPASE 9 had higher expression in groups I, A and I + A. The miRNAS analysis, miR-126 was higher in groups A and I + A, miR-155 was higher in groups I and I + A. CONCLUSIONS: We can conclude that apoptosis occurs in the cerebellar cortex, even if it is distant from the ischemic focus, and that miRNAs 126 and 155 present a correlation with cellular apoptosis in ischemic rats and submitted to the chronic alcohol model
 
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Publishing Date
2019-09-06
 
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