A Comparison between the Cytotoxicity Induced by Gossypol in Two Testicular Cell Lines

Background:Gossypol is a yellow toxic pigment from the cottonseed that can cause acute or chronic toxicity in humans and animals by affecting the testicular tissues. Nowadays cottonseed is used as food supplement for ruminants specially the sheep. In this study, two different stem cell lines of testicular tissue including GC1-spg (mouse testis) and SFTF-PI43 (sheep testis) cells were used to evaluation of gossypol cytotoxicity. Methods: The GC-1spg and the SFTF_PI43 cells were cultured in RPMI-1640 supplemented with fetal bovine serum (10%) and antibiotic (penicillin 105/ml, streptomycin100μg/ml), and then 5×104 cells/well were seeded in 24 well plates. Cultured cells were exposed to four different concentrations of gossypol (1.25, 2.5, 5 and 10μM). After 24 h incubation, cells viability test was performed using Trypan Blue dye exclusion and MTT assay. The Thiobarbituric Acid Reacting Substances (TBARS) and Ferric Reducing Activity Potential (FRAP) assays was performed on media. Result: In high concentrations (over than 2.5μM), Gossypol showed cytotoxic effects on cells. The IC50 for gossypol (using MTT assays) on SFTF-PI43 and GC-1spg cell lines was 2.2 μM and 3.2 μM, respectively. While the results for FRAP assay did not show any significant differences between the test and control groups, significantly higher lipid peroxidation was observed in SFTF-PI43 cells that were treated with higher doses of gossypol (10μM). Conclusion:In this research, we found that gossypol has cytotoxic effects on both examined testicular cell lines and increased lipid peroxidation, which is a probable mechanism of its toxicity on cell lines.