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Busulfan damages spermatogenic function by inducing orchitis

Lingjun Zhao1, Kaihui Wu1, Shiyuan Xu1,2

  • 1State Key Laboratory of Animal Biotech Breeding, Institute of Animal Science, Chinese Academy of Agricultural Sciences (CAAS), Beijing, China.

Plos One
|July 22, 2025
PubMed

Insights

Busulfan exposure damages male reproductive function by inducing testicular inflammation and disrupting the blood-testis barrier. This leads to reduced sperm quality and quantity, impacting fertility.

Area of Science:

  • Reproductive Toxicology
  • Cell Biology
  • Immunology

Background:

  • Busulfan is a chemotherapeutic agent with known reproductive toxicity.
  • The precise mechanisms by which busulfan impairs spermatogenesis remain incompletely understood.
  • Understanding these mechanisms is crucial for improving treatments for infertile patients and transplant recipients.

Purpose of the Study:

  • To elucidate the injury mechanism of busulfan on spermatogenic function in mice.
  • To investigate the effects of busulfan on testicular histology, sperm parameters, and inflammatory markers.
  • To examine the impact of busulfan on blood-testis barrier integrity and cytoskeletal components.

Main Methods:

  • Mice were treated with busulfan, and testicular and epididymal weights were measured.
  • Sperm concentration and morphology were analyzed.
  • Immunohistochemistry and immunofluorescence were used to assess testicular tissue.
  • Enzyme-linked immunosorbent assays (ELISAs) were performed to quantify serum inflammatory factors and blood-testis barrier-related proteins.

Main Results:

  • Busulfan treatment led to decreased testicular and epididymal weights, reduced sperm concentration, and increased sperm malformation.
  • Histological analysis revealed testicular interstitial cell infiltration, smaller seminiferous tubules, and disorganized spermatogenic cells.
  • Serum levels of inflammatory factors (IL-6, IL-1β, TNF-α) were significantly upregulated.
  • Levels of blood-testis barrier proteins (N-Cadherin, occludin, connexin 43) and vimentin decreased over time.

Conclusions:

  • Busulfan induces orchitis, disrupts the blood-testis barrier, and damages the spermatogenic microenvironment.
  • The observed decrease in vimentin indicates cytoskeletal damage, impairing Sertoli cell support for spermatogenic cells.
  • These disruptions negatively affect spermatogenesis and androgen regulation.
  • Findings offer insights to enhance the safety and efficacy of spermatogonial stem cell transplantation.

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