Examination of antioxidant effect of ellagic acid on testicular tissue in rats that were exposed to tobacco smoke

The disorders on cellular metabolism, lipid peroxidation, increase on membrane permeability, protein and DNA oxidation occurs because of the oxidative stress on tissues created by chemicals in tobacco smoke. It is known that exposure to tobacco smoke damages many tissues and organs. Toxic substances in tobacco smoke which are carried to every part of the body by blood, are likely to disturb the balance between oxidant - antioxidant systems on tissues. In this case, increased free radicals cause tissue damage. Ellagic acid is a polyphenol compound that found in fruits such as pomegranate, strawberry, blackberry, raspberry and grape. Its known that ellagic acid has protective effects against oxidative damage. Also its anti-cancerogenic, anti-apoptotic and anti-inflammatory effects are known. In our study, it is aimed to examine the protective effects of ellagic acid, which is a powerful anti-oxidant, against oxidative damage on testis tissue caused by the chemicals in tobacco smoke. In this study, 8 weeks old and average 200±10 grams weight twenty-four male Spraque-Dawley rats were used. The rats were divided randomly into 4 equal groups: group I (Control), group II (Tobacco smoke), group III (Tobacco smoke + corn oil) and group IV (Tobacco smoke + ellagic acid). The rats in group II, III and IV were exposed to tobacco smoke 1 hour twice a day. In addition to tobacco smoke exposure, 12 mg/kg and dissolved in corn oil ellagic acid, was applied to the rats in group IV by oral gavage. Equal amount of corn oil used in solving ellagic acid was applied to the rats by oral gavage in group III. At the end of the 12 weeks experimental period rats were decapitated. Testicular tissues were removed and processed by using routine parafine techniques. The testis slides were stained by H&E, PAS and Masson's Trichrome methods. Also endothelial nitric oxide synthase (eNOS) immunohistochemical stain and TUNEL methods for apoptosis were applied. Biochemical analyzes on blood samples and sperm analyzes on sperm samples were performed. A significant decrease of percentage of body weight change was detected in tobacco smoke and tobacco smoke + corn oil groups when compared with control group. There was no significantly difference between tobacco smoke + ellagic acid and the other groups in terms percentage of body weight change. Right and left testis weights of rats in tobacco smoke + ellagic acid group, significantly increased compared with control group. There was no statistically difference detected in epididymis, prostate and seminal vesicle weights between the groups. The relative testis weights of rats in tobacco smoke + ellagic acid group were significant increased compared to the control group. The difference of sperm motility and sperm concentration between the groups wasn't found significant. A significant increase in abnormal sperm amount was detected in tobacco smoke group compared with control group. The reduction in the amount of abnormal sperm was found in tobacco smoke + ellagic acid group compared with tobacco smoke group. In histological examination with light microscope of rat testis tissues in tobacco smoke group compared with control group; degeneration in seminiferious tubule germinative epitelium, seperation in basement membranes, vascular congestion, atrophic tubules, edema in interstitial area and immature cell debris in tubule lumen were detected. On the other hand, in the rat testis which were experimented tobacco smoke + corn oil, similiar finding with the tobacco smoke group were detected. Significant improvement were observed in germinative epitelium degeneration, seperation of seminiferious tubule basement membranes, vascular congestion and intersititial edema in rat testis tissues of tobacco smoke + ellagic acid group. In tobacco smoke and tobacco smoke + corn oil groups, a significant increase was detected in TUNEL positive cells number compared to the control group. In tobacco smoke + ellagic acid group, the number of TUNEL positive cells was detected similar to control group. A significantly increased eNOS immunoreactivity was observed in tobacco smoke and tobacco smoke + corn oil group compared to the control group. On the other hand, eNOS immunoreactivity decreased in tobacco smoke + ellagic acid group. A significantly increase was detected in the levels of MDA, in tobacco smoke and tobacco smoke + corn oil groups compared with the control group. It was found that ellagic acid administration reduced the MDA levels similar to the control group. CAT and GSH-Px enzyme activities in tobacco smoke and tobacco smoke + corn oil groups were significantly decreased compared to the control group. Also significantly increase in CAT and GSH-Px enzyme activities was detected in tobacco smoke + ellagic acid group compared with the tobacco smoke and tobacco smoke + corn oil groups. As a result of this experimental study, it was observed that exposure to tobacco smoke effects negatively the male reproductive system. it was also observed that ellagic acid, which is used as a protector against tobacco smoke exposure, decreases the negative effects of tobacco smoke prominently by means of its antioxidant property.