Evaluation of immunological parameters of the epididymis of diacerein-treated rats

• Grant number: 24/08088-9
• Support Opportunities: Scholarships in Brazil - Scientific Initiation
• Start date: July 01, 2024
• End date: June 30, 2025
• Field of knowledge: Biological Sciences - Biology
• Principal Investigator: Estela Sasso Cerri
• Grantee: Laryssa Morandine Macas
• Host Institution: Faculdade de Odontologia (FOAr). Universidade Estadual Paulista (UNESP). Campus de Araraquara. Araraquara , SP, Brazil

Abstract

The epididymis plays a crucial role in male fertility. During the passage along the epididymis, spermatozoa, newly released from the testis, acquire ability to fertilize the oocyte. This organ consists of a single highly convoluted duct divided into four main anatomical regions: the initial segment, head, body, and tail. The latter region is important for the final maturation, storage, and protection of sperm until ejaculation. The epididymis is an immune-privileged organ, composed of a complex cellular environment that includes macrophages, dendritic cells, and lymphocytes, maintaining a balance between pro-inflammatory cytokines such as TNF-± and IL-1² and anti-inflammatory cytokines such as IL-10. Diacerein is an anti-inflammatory drug that inhibits the pro-inflammatory cytokines. Treatment of rats with this drug has caused structural changes in testis and reduction of testosterone synthesis. Since epididymis is an androgen-dependent organ that expresses an immunological profile, the aim of this study is to evaluate the epididymal structural integrity and the immune cells population of epididymal epithelium and stroma following the diacerein inhibitory effect on the pro-inflammatory cytokines. Ten Holtzman male rats will be grouped into: control group (CG; n = 5) and diacerein group (DG; n = 5). The DG animals will receive 100 mg/kg of diacerein daily by gavage for 30 days while the CG animals will receive only distilled water for the same period. The epididymides will be fixed and processed for paraffin and historesin embedding for morphological analyses and immunolocalization of IL-1² and TNF-±. Immunofluorescence reactions will be performed for detection of F4/80 (mononuclear phagocyte marker), CD4+ and CD8+ (T lymphocyte markers), and IL-10, and the immunofluorescent areas will be measured. Moreover, halo cells and mast cells will also be quantified in the epithelial and stromal tissue, respectively. The differences between groups will be statistically analyzed by Student's t-test (pd0.05).