A Histological Study of the Effects of Administration and Withdrawal of Anabolic Androgenic Steroid (Nandrolone Decanoate) on the Hippocampus and the Prefrontal Cortex in Adult Male Albino Rats
Authors: Mohamed Amera Taha , Soliman Younus Azza Salah El Din , Rady Hagar Yousry , Elmoneim Marwa Abd 
Published: July 2024

Abstract

Background
The PFC and the hippocampus are parts of the limbic systems and have a major role in emotion regulation, memory and learning. Anabolic androgenic steroids (AASs) are large groups of synthetic derivatives of testosterone, prescribed for the treatment of male hypogonadism. Common abuse of AASs has spread among the general population and

Aim of the Work
The aim of the current work was to study the effects of AASs on the PFC and the hippocampus histologically and statistically and to evaluate the reversibility of these effects in these regions after AASs withdrawal.

Material and Methods
24 adult male albino rats (3 – 6) months old (weighing 200-225 gm) were used in the current study and divided into 3 groups; according to the procedure and treatment type: control group (I) (12 rats) and AASs treated group (II) (6 rats) and Withdrawal group (III) (6 rats). Fixation for the brain using bouin solution was done first in situ followed by its removal and transfer to a glass bottle containing bouin solution for 24 hours to ensure complete fixation. The PFC and the hippocampi were obtained through parasagittal section. Specimens were processed for preparation of paraffin blocks then stained with H&E stain and immunohistochemically stained for GFAP antibody. Image analysis &morphometry were performed on stained sections for; number of surviving pyramidal neurons and the mean area percentage of GFAP staining.

Results
On examination of H&E-stained sections of control groups, the PFC was formed of the six- layers, the molecular layer, outer granular layer, outer pyramidal layer, inner granular layer, inner pyramidal layer and the multiform layer. The Hippocampus was formed of two components: the dentate gyrus (DG), the hippocampus proper (Cornu Ammonis, with its four regions: CA1, CA2, CA3 and CA4). The cells forming the hippocampus proper were arranged in three layers; the molecular, the pyramidal and the polymorphic layers. In immunohistochemically stained section, the present work showed minimal apparent immune-reaction in the cytoplasm and processes of astrocytes in the PFC and CA1 region of the hippocampus in the control group. On examination of H&E-stained sections of the AASs treated groups, signs of neuronal damage were observed. The inner pyramidal layer of the PFC and the pyramidal layer of CA1 region of the hippocampus revealed abnormal shrunken pyramidal cells having deeply stained nuclei, peri-cellular spaces and flame like appearance with pointed end. Moreover, these layers demonstrated also normal pyramidal cells showing also large rounded open vesicular nuclei and prominent nucleoli. Immunohistochemically stained sections of the treated group in both regions showed apparent extensive brownish immunostaining of the astrocytes with ramifying processes along the layer. Examination of H&E-stained sections of the withdrawal group revealed the inner pyramidal layer of the PFC and the pyramidal layer of CA1 region of the hippocampus showing normal pyramidal cells having large rounded open vesicular nuclei, prominent nucleoli and apical dendrites. It also showed dark irregular cells having deeply stained nuclei, peri- cellular spaces and flame like appearance with pointed end. Immunohistochemically stained section of the treated group in both regions showed apparent minimal brownish immunostaining of the astrocytes.

Conclusion
Results of the present experimental study demonstrated that AASs (in the form of ND) administration affecting the PFC and the hippocampus resulted in several histopathological and morphometric harmful effects which regressed to a lesser extent through its withdrawal after 2 weeks.