The effects of Di-N-Buthyl phthalate (DBP) on testis and various microscopic investigation of Resveratrol's possible protective features

In this study it was aimed to observe the possible protective effects of Resveratrol which is preferable as a antioxidant against the damage of an endocrine disruptor di n butyl phthalate on testicles with the help of immunohistochemical methods.36 male albino Wistar rats,which are 20 days old are used.They divided into 6 seperate groups.Each group has 6 experimental subjects. I. Group: Control, II. Group:Solvent (Carboxymethyl cellulose,CMC 10 ml/kg), III.Group : 500 mg/kg/day DBP , IV.Group : 500 mg/kg/day DBP+20 mg/kg/day Resveratrol ,V. Group: 1000 mg/kg/day DBP, VI. Group : 1000 mg/kg/day +20 mg/kg/day Resveratrol. At the end of this time,after gathered testicle tissues were observed with ligth microscope paraffin blocks were prepared.C-kit was used on sections that are taken blocks to observe proliferation, AT1 used in order to observe the stress rate of seminiferous epithel especially Leydig cells and immunohistochemical stainig with estrogen receptor ? anticors used to observe whether interaction was at hormonal level or not.TUNEL method to sembolize apoptosis,was used.Spermatogenetic epithel length are measured at 6 area for 6 experimental subjects per group. Shapiro Wilk Test, Kruskal Wallis Test with Bonferroni correction and Mann Whitney U Test with Bonferroni correction are used.In c-kit immune staining,at groups that are treated with 500mg/kg and 1000 mg/kg DBP,depending on increasing DBP dose, the involvements decrease extremely in contrast to control and CMC applied sham control groups.On the other hand,at 500mg/kg DBP+Resveratrol applied groups conversely to other groups,involvements are similar to control group. At 1000 mg/kg DBP+Resveratrol applied group,it was observed that Resvaratrol was not enough.The rise of the involvement of AT1 at 500 mg/kg and 1000 mg/kg DBP treated groups;at Resveratrol applied groups partially return of the negative effect was observed;but involvement of Leydig cells at high dose of DBP and Resveratrol treated groups was still preserved.When effect of DBP on testicles and estrogen receptor level are examined,although significant differences between groups are not seen,It is observed that high doses of DBP decreases estrogen level and It is also observed that Resveratrol is not enough in endocrin protection ability.Observation made with the TUNEL method,it was determined that especially high-dose of DBP increased apoptosis in all cell types.Resveratrol application in the apoptotic process,it showed that the protective effect with low-dose DBP but not prevention apoptosis with high-dose application.Also the statistical evaluation indicated that, in contrast to other groups, at DBP treated groups ,epithelium height and diameter of seminiferous tubules decreased but this reduction were returned back at Resveratrol applied group.As a result,these lead us to conclusion that the increasing application of DBP dose negatively affects the cell cycle of the testical structure parallelly, and Resveratrol can show cyclic protective effect against hormonal and cyclic changes but this is not fully adequate and there is no hormonal effects.