Role of mitochondria in ciprofloxacin-induced apoptosis in murine sperm cells

Ciprofloxacin (CPFX) has been reported to inhibit cell growth and induce apoptosis in certain eukaryotic cells. The role of
the mitochondrial pathway in CPFX-induced apoptosis in cultured murine sperm cells was investigated. Methods and Results:
Spermcells (5�103 cells/well) from 8-week-oldNMRI malemice were cultured in 150 mL of HAM’s F10with 25mmol/L (4-(2-hydroxyethyl)-
1-piperazineethanesulfonic acid ) HEPES and 10% human serum albumin and were incubated with 50, 100, 200, 400, and 800
mg/mL CPFX for 24 and 36 hours. Cell cytotoxicity, mitochondrial membrane potential (DCM), and concentrations of caspase 3 and
caspase 9 were assessed in CPFX-treated cultured murine sperm cells by MTT (3-(4,5-dimethylthiazol-2-Yl)-2,5-diphenyltetrazolium
bromide) , JC-1 (5, 5a´, 6, 6a´-tetrachloro-1, 1a´, 3, 3a´-tetraethylbenzimidazol-carbocyanine iodide)aggregation, and caspase 3 and caspase
9 assays, respectively. Increasing doses of CPFX showed significant cytotoxicity (EC50 ¼ 146.73 mg/mL). Significant loss of DCm
was observed in spermcells treatedwith�50mg/mLCPFXfor 36 hours. Significant increases in caspase 9 and caspase 3 concentrations
were also found in cells treated with �50 mg/mL CPFX for 24 and 36 hours, respectively (P < .001). Conclusions: Effects of clinically
reachable doses of CPFX on cultured murine sperm cellswere investigated and revealed that it may cause sperm cell toxicity by induction
of apoptosis through the mitochondrial pathway in the clinically reachable concentrations.