MORPHOLOGICAL ANALYSIS AND APOPTOSIS EXPRESSION IN EXPERIMENTAL CEREBRAL ISCHEMIA ASSOCIATED WITH ALCOHOLISM MODEL
Authors: da Silva Jairo Pinheiro , da Cunha Tirapelli Daniela Pretti , de Fátima Galli Tazima Maria , de Assis Cirino Mucio Luiz , Neto Francisco Ribeiro Tavares , Novais Paulo Cezar , Bisson Gabriela Silva , Neto Fermino Sanzes Lizarte , Colli Benedito Oscar , Junior Carlos Gilberto Carlotti , Tirapelli Luis Fernando 
Published: June 2021

Introduction

Brain ischemia is a major cause of death and neurologic disability in many countries. It is a frequent clinical condition of difficult therapeutic solution mainly because its physiopathological studies are embarrassed by factors like the diversity of the anatomical location of the lesions, its etiology and clinical signs and symptoms. The alcoholism is a health problemin many societies,larger studies are needed on effects of alcohol associated with cerebral ischemia. It is known that neurons can die few days after stroke and some researchers suggest that apoptosis is the main physiopathological mechanism involved in delayed neuronal death.

Objectives

To evaluate the histopathological changes, morphometry, expression of apoptosis‐related proteins CASPASE3 and BCL2, serum expression of miR‐21 after experimental cerebral ischemia followed by reperfusion for 48 hours, with or without a model of chronic alcoholism.

Material and Methods

We used 50 Wistar rats, divided into 5 experimental groups: Control: only anesthetic procedure; Sham: complete simulation of the surgical procedure; Ischemia (I): focal cerebral ischemia for 90 minutes + reperfusion for 48 hours; Alcoholic(A): received daily absolute ethyl alcohol diluted to 20% in water for four weeks, and, Alcoholic and Ischemic (I + A): the same treatment in group A and I. The brain samples collected were processed for histopathological analysis (by light microscopy and transmission electron microscopy), morphometric (counting of neurons in the striatum), both by the technique of luxol fast blue, and immunohistochemistry (for protein expression of CASPASE3 and BCL2). The blood of the ventral tail artery was collected for analysis of expression of miR‐21 by qPCR.

Results

We observed histopathological changes in animals with a higher degree of ischemic and ischemic groups associated with alcoholism in three areas analyzed: swelling of neuronal cytoplasm, foamy nucleus, neural cells with pyknotic nuclei, interstitial edema and foci of inflammatory infiltrate. The neuronal loss was higher in the medial striatum of animals in groups I and I+A. The protein expression of CASPASE3 was higher than the expression of BCL2, especially in groups I and I + A for the two proteins. The expression of CASPASE3 was higher in the region corresponding to the penumbra. Histopathology revealed that the ischemic focus had a greater number of pyknotic nuclei suggestive of necrosis.

Conclusions

The histopathologic and morphometric changes observed mainly in the ischemic focus were correlated with the expression of CASPASE3, the greater the area of penumbra, and were more severe in the ischemic animals when associated with chronic alcoholism. The expression of BCL2 was slightly higher where histopathological changes and expression of CASPASE3 were less evident. The serum expression of miR‐21 was not indicative of ischemic and/or associated with chronic alcoholism.

This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.