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This study tested the hypothesis that different doses of nandrolone decanoate ND will cause changes in the estrous cycle and ovarian tissue of adult rats; and investigated the duration of the recovery period that is sufficient to restore the damage in the animals treated with different doses. Wistar rats were treated with ND at doses of 1. All animals were divided into three groups according to the treatment periods: Estrous cycle was monitored daily, and at the end of each period, the animals were euthanized for histopathological analysis.
The prescription and commercialization of these drugs are strictly controlled in many countries. Although deleterious effects promoted by AAS are well understood, few studies have focused attention to their effects on female reproductive morphological parameters. The estrous cycle is a periodic physiologic process, with an average duration of 4.
This study brings novel information regarding the reversibility of some effects promoted by ND in the reproductive cycle and ovaries. The experimental protocol followed the ethical principles in animal research adopted by the Brazilian College of Animal Experimentation and was approved by the Ethical Committee for Animals Use Permit number: Both CE and CD groups received subcutaneous s.
The first injection was administered at the estrus phase of the cycle in all animals. In this regard, the evaluation of ovarian tissue was carried out in a specific stage of the estrous cycle that androgenized females persisted or maintained for a long time during the experimental period. At the end of each period, the females who had regular estrous cyclicity were euthanized in estrus phase, while the acyclic females were euthanized at the cycle phase wherein persisted.
For both cases, the ovaries were collected and processed for histopathological analysis. Chronological schedule for the experimental design in days.
The phases of the estrous cycle were identified as follows: All of the histological samples were handled in a blinded fashion. The criterion for the identification of atretic follicles was based on the presence of three characteristics: Then, sections were counterstained with haematoxylin. Negative controls were made by omitting the primary antibody. The results were analysed under a Zeiss Axiophot II microscope Carl Zeiss, Oberkochen, Germany based on the levels of staining intensity as absent, weak, moderate and strong reaction.
For negative immunolabelling, no primary antibody was added. The data were analysed by parametric analysis of variance anova followed by Tukey's test, or by nonparametric Kruskal—Wallis test followed by Student—Newman—Keuls test, according to the characteristics of each variable. Statistical analysis was performed using graphpad prism software, version 5. Females treated with lower doses 1. At the end of this period, the rats restored the cycle, but irregularly.
The group treated with 7. Due to estrous cycle restoration, these groups 1. Control group with regular cycles was euthanized in the estrus phase CE. Data are expressed as the median and interquartile deviation. Notably, the group treated with 1. These cells presented polymorphonuclear aspect with prominent nucleolus, and abundant and pallid cytoplasm. In the group treated with 7.
Stained with Mallory's trichrome c, f, g, h, i, j, k, l, n and Van Gieson a, b, d, e, m. Stained with Mallory's trichrome a—c and Orcein d. Conversely, the animals treated with higher doses of ND 7. This result occurred after the treatment and recovery periods. Stained with Mallory's trichrome a and haematoxylin b, c. The area of these follicles remained reduced in the groups treated with 1.
In this study, the estrous cycle and ovarian structures were affected by treatment with ND in all of the doses, and recovery periods did not restore the adverse effects, mainly when the steroid was administered at the highest dose level. The absence of changes in the ovarian weight after ND treatment probably occurred due to the presence of corpora lutea in the ovaries, even with a decrease in the population of follicles. In the groups treated with highest doses, the ovarian weight remained unchanged.
Moreover, these hormonal disturbances may affect the regulation of the estrous cycle as observed. Thus, it can be considered that these females showed a physiological anestrus. The animals treated with the lower doses exhibited an irregular cycle at the end of the period, with extension of diestrus. This study described for the first time the histopathological and morphometrical changes in the ovarian tissue arising from different doses of ND and varying according to the recovery periods.
In general, both folliculogenesis and luteogenesis have been severely damaged by ND, indicating that neither doses nor period of treatment are safe when administered in therapeutic or supraphysiological levels. The lowest dose of ND 1. These effects may be caused indirectly via the neuroendocrine axis or even directly via androgen receptors AR expressed in the granulosa and theca cells Drummond An explanation for this is provided by the presence of oestrogen receptors in the rat corpora lutea CL.
However, oestrogen is not always required for the action of androgens in the CL. In animals receiving doses of 1. These results suggest that the action of androstenedione on luteal progesterone production and secretion at the time of luteolysis occurs through an androgenic mechanism. The presence of stromal fibrosis in the different groups was due to tissue degeneration.
This effect has been extensively reported in animal or human coronary arteries. In the present study, vasodilation observed in ovarian medulla occurred only at the dose of 7. The changes in the features of ovarian tissue, associated with a decrease in the population of follicular units, are capable of promoting a further significant reproductive loss for the animal, considering that both preservation and function of the follicles are damaged.
Only two females receiving 1. Based on these results, the current study suggested that the treatment of ND is able to affect ovulation. Such structures were scarce in the most groups varying from 1 to 2 anomalous follicles and absent in the group treated with 3. This positive reaction was reduced after a longer recovery period, probably due to a downregulation of AR. In summary, all the different doses of ND administered 1. National Center for Biotechnology Information , U.
Int J Exp Pathol. Published online Nov Received Feb 9; Accepted Jul Summary This study tested the hypothesis that different doses of nandrolone decanoate ND will cause changes in the estrous cycle and ovarian tissue of adult rats; and investigated the duration of the recovery period that is sufficient to restore the damage in the animals treated with different doses. Open in a separate window. Statistical analysis The data were analysed by parametric analysis of variance anova followed by Tukey's test, or by nonparametric Kruskal—Wallis test followed by Student—Newman—Keuls test, according to the characteristics of each variable.
X Presence; — Absence. Discussion In this study, the estrous cycle and ovarian structures were affected by treatment with ND in all of the doses, and recovery periods did not restore the adverse effects, mainly when the steroid was administered at the highest dose level.
Conflict of interest The authors declare no conflict of interest. Gerontology 22 , — Contraception 72 , 77— Endocrinology , — Anaesthesia 60 , — Micron 37 , — Sports 21 , e91—e Potential mechanisms of ovarian injury.