The effect of Brazilian propolis and ametryn on Leydig cells and testosterone production of Wistar rats

Brazilian green propolis is an important chemical defense used by bees against pathogenic microorganisms. In popular medicine propolis have been used as a remedy in wound treatment, stomach ulcers, mouth rinse and in the preparation of food and beverages. The indiscriminate use of propolis can cause damages in male gonads. Preview studies showed that propolis can increase sperm production. Ametryn is a triazine herbicide that inhibits photosynthesis in plants and it is used in many countries as a selective herbicide to control broadleaf and invasive grasses in corn and sugarcane crops. Wistar rats treated with ametryn showed decrease in Leydig cell proportion such as and sperm decrease. Studies show the effect of these two compounds (ametryn and propolis) applied separately in Wistar rat testis. The aim of the present work was investigate the effect of concomitant application of these two compounds on Leydig cells morphology and quantify PSA and testosterone hormones in blood. Leydig cells are located in the interstitial region of the testis and are responsible for testosterone production. The treatment was performed for 90 days old-male Wistar rats (Rattus norvegicus), administered by gavage. The groups were divided into four groups (n = 6) C: control group (5mg of water/day), P: propolis group (6mg/kg/day of propolis extract), A: ametryn group (15mg/kg/day os ametryn) and P+A: propolis and ametryn group (association of propolis and ametryn doses). They were anaesthetized, euthanized and perfused after 56 days, before perfusion, blood was collected by cardiac puncture. Testis were fixed with Karnovsky and routinely processed for paraffin inclusion. The blood analysis were performed by Sao Lucas Laboratory, in Rio Claro, SP. Leydig cells stereology showed significant differences comparing P+A group and the other groups. The proportion of cell nucleus increased and cytoplasm proportion decreased. In the blood analysis, testosterone hormone production decreased significantly in the P+A group compared to the other groups (P < 0.0001) and increased in the ametryn group compared to the propolis group (P = 0,0395). In previous investigations of our research group with different propolis doses, all exposed groups showed a decrease in nuclear proportion and increased Leydig cell cytoplasm proportion. According to previous studies did not observe any change in the relative proportions of nucleus and cytoplasm with Ametryn treatment. Therefore, our results show that Propolis along with Ametryn interfered in the composition nucleus/cytoplasm of Leydig cells, consequently in testosterone production, which may negatively affect spermatogenesis. Further analysis such as spertmatides count in testis are necessary to confirm these results.