Effect of human recombinant granulocyte colony-stimulating factor on rat busulfan-induced testis injury

J Mol Histol. 2016 Feb;47(1):59-67. doi: 10.1007/s10735-015-9647-y. Epub 2015 Dec 29.

Abstract

Granulocyte colony-stimulating factor (G-CSF) is known for proliferation and anti-apoptotic activities. We aimed to use this growth factor in busulfan-injured testis. 32 male Wistar rats were injected with a double dosage of 15 ml/kg busulfan with 14 days interval. Administration of human recombinant G-CSF (100 µg/kg) subcutaneously was performed in two different time periods: 3 days before and 2 days after receiving busulfan, G-CSF1; and at days 14-18 of busulfan injection, G-CSF2. Animals were sacrificed at the end of week five. Histological analysis, testis weight and sperm parameters (sperm count and viability) has been checked. Expressions of DEAD (Asp-Glu-Ala-Asp) box polypeptide 4 (DDX4), deleted in azoospermia like (DAZL), transition protein 2 (TP2), proliferating cell nuclear antigen (PCNA) and 5-Bromo-20-deoxyuridine (BrdU) were assessed. Empty seminiferous tubules were apparent in the busulfan- and G-CSF2-injected rats, but not in the G-CSF1 group. The G-CSF1-treated animals showed an increase in testis weight and sperm count and viability along with high expressions of DDX4, DAZL, TP2, PCNA and BrdU; even so, the changes were reversed in the busulfan and G-CSF2 groups (for all p < 0.05). Our results revealed that G-CSF application prior to busulfan insult is a promising approach in fertility maintenance.

Keywords: Busulfan; Granulocyte colony-stimulating factor; Infertility; Spermatogenesis.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Bromodeoxyuridine / metabolism
  • Busulfan / toxicity*
  • Cell Survival / drug effects
  • DEAD-box RNA Helicases / genetics
  • DEAD-box RNA Helicases / metabolism
  • DNA-Binding Proteins
  • Gene Expression Regulation
  • Granulocyte Colony-Stimulating Factor / pharmacology*
  • Injections, Subcutaneous
  • Male
  • Nuclear Proteins / genetics
  • Nuclear Proteins / metabolism
  • Organ Size / drug effects
  • Proliferating Cell Nuclear Antigen / genetics
  • Proliferating Cell Nuclear Antigen / metabolism
  • RNA-Binding Proteins / genetics
  • RNA-Binding Proteins / metabolism
  • Rats
  • Rats, Wistar
  • Recombinant Proteins / pharmacology
  • Semen Analysis
  • Signal Transduction
  • Sperm Count
  • Spermatogenesis / drug effects*
  • Testis / drug effects*
  • Testis / metabolism
  • Testis / pathology
  • Testis / ultrastructure

Substances

  • DAZL protein, rat
  • DNA-Binding Proteins
  • Nuclear Proteins
  • Proliferating Cell Nuclear Antigen
  • RNA-Binding Proteins
  • Recombinant Proteins
  • Tnp2 protein, rat
  • Granulocyte Colony-Stimulating Factor
  • Ddx4 protein, rat
  • DEAD-box RNA Helicases
  • Busulfan
  • Bromodeoxyuridine