Ciprofloxacin Induces Toxic and Apoptotic Effects on Skeletal Muscles

Introduction: Ciprofloxacin is a second-generation fluoroquinolone antibiotic commonly used in therapy of many microbial infections. Rhabdomyolis is one of the serious side effects of ciprofloxacin. The use of ciprofloxacin is associated with a raised risk of myopathy especially in patients taking statins, but the mechanisms underlying ciprofloxacin-induced myopathy are poorly known. In this study, we investigated the effects of ciprofloxacin on isolated rat skeletal muscles mitochondria.Methods: Rat skeletal muscles mitochondria were obtained by differential ultracentrifugation and incubated with different concentrations of ciprofloxacin (35, 70 and 140 μM) .The activity of mitochondrial complex II was assayed via the measurement of MTT reduction. The mitochondrial ROS measurement was perfomed using the fluorescent probe DCFH-DA. The Rhodamine 123 (Rh 123) redistribution technique was used for MMP measurement. Mitochondrial swelling was measured spectrophotometrically in duration 1 hour. The concentration of cytochrome c was determined through using the Quantikine Rat/mouse cytochrome c Immunoassay kit. Data were analyzed using the Graph pad prism software, version 6.Results: The results showed that IC50 of ciprofloxacin was assigned approximately 70 μM. Under our experimental conditions, succinate dehydrogenase activity was significantly reduced within 60 min incubation with different concentrations (35, 70 and 140 μM) of ciprofloxacin. In Our study showed that H2O2 production was increased in a time and concentration-dependent. According to our results all concentrations of ciprofloxacin after 60 min incubation, significantly decreased MMP in a time-dependent manner. Our results showed that ciprofloxacin at all concentrations (35, 70 and 140 μM) following 60 min incubation with skeletal muscles mitochondria significantly (P<0.05) increased the mitochondria swelling, compared to untreated control groups. Ciprofloxacin (35, 70 and 140 μM) caused a significant (P<0.05) decrease in the complex IV activity. Our results demonstrated that ciprofloxacin induced a rise in mitochondrial reactive species (ROS) formation and mitochondrial membrane potential (MMP) collapse before mitochondrial swelling ensued in isolated skeletal muscles mitochondria. In addition collapse of MMP and mitochondrial swelling produced release of cytochrome c via outer membrane rupture or mitochondrial permeability transition (MPT) pore opening.Conclusion: According to the results, we suggested that ciprofloxacin- induced myopathy is the result of a disruptive effect on mitochondrial respiratory chain and induction of ROS-mediated apoptosis signaling in skeletal muscle cells.

Keywords:

Ciprofloxacin , Mitochondria , Myopathy , ROS formation , Toxicity