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:
Sperm cells (5×103 cells/well) from 8-week-old NMRI male mice were cultured in 150 μL of HAM’s F10 with 25 mmol/L (4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid ) HEPES and 10% human serum albumin and were incubated with 50, 100, 200, 400, and 800 µg/mL CPFX for 24 and 36 hours. Cell cytotoxicity, mitochondrial membrane potential (ΔΨM), 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, 5á, 6, 6á-tetrachloro-1, 1á, 3, 3á-tetraethylbenzimidazol-carbocyanine iodide)aggregation, and caspase 3 and caspase 9 assays, respectively. Increasing doses of CPFX showed significant cytotoxicity (EC50 = 146.73 μg/mL). Significant loss of ΔΨm was observed in sperm cells treated with ≥50 μg/mL CPFX for 36 hours. Significant increases in caspase 9 and caspase 3 concentrations were also found in cells treated with ≥50 μg/mL CPFX for 24 and 36 hours, respectively (P < .001).
Conclusions:
Effects of clinically reachable doses of CPFX on cultured murine sperm cells were investigated and revealed that it may cause sperm cell toxicity by induction of apoptosis through the mitochondrial pathway in the clinically reachable concentrations.
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