Protective role of antioxidants in experimental chronic alcohol intoxication

Abstract

Alcoholic disease that develops as a result of the systematic use of alcohol is always accompanied with a violation of the functions of vital organs. In numerous publications, the effect of systematic alcohol abuse on the liver, brain, heart, and gastrointestinal tract has been demonstrated [1, 2]. At the same time, little attention is paid to the change in the structure of organs such as the adrenal glands under the influence of the chronic use of ethanol.

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RATIONALE Alcoholic disease that develops as a result of the systematic use of alcohol is always accompanied with a violation of the functions of vital organs. In numerous publications, the effect of systematic alcohol abuse on the liver, brain, heart, and gastrointestinal tract has been demonstrated [1, 2]. At the same time, little attention is paid to the change in the structure of organs such as the adrenal glands under the influence of the chronic use of ethanol. PURPOSE Study the influence of antioxidant drugs on adrenal medulla histology. METHODS Experimental studies were conducted in the autumn-winter period. In our experiment, we used 30 Wistar albino male rats weighing 200-235 g. divided into the following groups: control and 4 experimental groups. After14 days of quarantine, rats in experimental groups were undergone to alcohol intoxication during 60 days. Group one received alcohol only. Group two received oral alfa-tocoferol (day 41-day 60). Groups three and four received squalene - tocoferol composition with the final volume of 1 or 2 ml of composition per one rat correspondingly. For morphological studies of the adrenal glands of animals at the end of the experimental period under ether anesthesia were decapitated, the abdominal cavity was opened, then right and left adrenal glands were removed and weighed on torsion scales. One part of the excised tissue of the adrenal glands was fixed in 10% neutral formalin, the other - within 24 hours in Bouin liquid. After routine dehydration, tissue blocks were poured into the histomix casting. Morphological studies were performed on serial paraffin sections with a thickness of 6μm. Sections were stained with hematoxylin and eosin for general histological morphometric studies. After staining, the sections were dehydrated in alcohols of rising concentration, clarified in meta-xylene and placed in Canadian balsam [3]. The state of the cortical substance was assessed by visual microscopy and morphometry, during which the width of the cortex and the volume of the nuclei were measured. To determine the ratio of cortical and medulla, as well as to determine the width of the zones of the cortex of the adrenal glands, an eyepiece micrometer was used. Karyometric studies were also conducted, for which the diameter of 100 nuclei for each animal was measured using an eyepiece micrometer. RESULTS In the micro preparations of the control group of animals stained with hematoxylin and eosin, three zones of the cortex were determined: glomerular, beam, reticular, oriented from the gland capsule to the medulla. When analyzing the width of the zones of the cortex, it was found that the longest is the beam zone (554.2 ± 31.0 μm), its mesh zone is 100 times (100.8 ± 9.1 μm) and almost six times the glomerular zone ( 85.0 ± 4.5 μm). These differences may indicate the existence of features of the functioning of cells in individual zones of the cortex. The parenchyma of the gland is enclosed in a connective tissue capsule. Within the glomerular zone, cells are polymorphic, of various sizes, in most cases of relatively small size (90.8 ± 5.9 μm2). The subcutaneous, transitional zone with a relatively dense granular cytoplasm is expressed. Nucleolemma along the perimeter is flat. In the center of the nucleus there is determined stained nucleolus. The second group of animals received exogenous alcohol and metabolic correction using the antioxidant α-tocopherol. In this group the adrenal cortex width was wider compared to the control, which is associated with an increase in the beam zone (656.6 ± 22.0), the width of the glomerular zone decreased in comparison with the control (76.1 ± 3.2), and the values of the width of the mesh zone did not differ significantly from the control (100.2 ± 6.7). We analyzed the dependence of the cortical zone width, nucleus size, cell cytoplasm, nuclear cytoplasmic index and histochemical markers of protein content in the nucleus and cytoplasm and RNA in the nucleolus and cytoplasm. It was estimated that a decrease in the glomerular cortical zone in experimental animals was accompanied by a decrease in the cell cross-sectional area with a simultaneous decrease in the protein content in the cytoplasm, and these parameters were in close correlation. A statistically significant increase in the core volume was also detected. CONCLUSION The use of the antioxidant α-tocopherol in combination with squalene made it possible to bring histochemical parameters closer to the values of the control group of animals that also received α-tocopherol, as well as intact control animals. The use of antioxidants could be promising for the correction of the alcohol-induced disorders.