2-Bromopropane induces DNA damage, impairs functional antioxidant cellular defenses, and enhances the lipid peroxidation process in primary cultures of rat Leydig cells.

Utilization of highly enriched preparations of steroidogenic Leydig cells has proven invaluable for studying the direct effects of various hormones and agents on Leydig cell function in vitro. It is widely reported that male reproductive organs are particularly susceptible to the deleterious effects of reactive oxygen species (ROS) and lipid peroxidation, which ultimately lead to impaired fertility. The purpose of the study was to examine the potential of 2-bromopropane (2-BP) to induce oxidative stress and antioxidant function in primary cultures of rat Leydig cells. Leydig cells were isolated from the testes of Sprague-Dawley rats. The purity of Leydig cells was determined to be 94.6% and the cells maintained their testosterone secreting capabilities for 48 h. Fresh medium containing 2-BP (1.00, 0.10, 0.01 mM, and vehicle control) and 1 U human chorionic gonadotropin (hCG) were added in the cell culture. Superoxide dismutase (SOD) activity, malondialdehyde (MDA), and glutathione peroxidase (GSH-PX) were analyzed in the medium of each well by biochemical methods. Additionally, DNA damage was examined using the Comet assay. The proportion of cells with undamaged DNA was decreased significantly and those with different grades of damaged DNA were increased significantly in the cells exposed to 2-BP. The level of MDA and GSH-PX activity increased significantly in the cell groups exposed to 0.10 and 1.00 mM 2-BP, whereas, SOD activity decreased considerably in these two groups of cells when compared to the control. The data indicate that 2-BP induces DNA damage, impairs functional antioxidant cellular defenses, and enhances the lipid peroxidation in cultured Leydig cells. These effects may be responsible for the testicular toxicity noted in laboratory animals and humans. Copyright 2002 Elsevier Science Inc.