1,8-Cineol (Eucalyptol) Disrupts Membrane Integrity and Induces Oxidative Stress in Methicillin-Resistant Staphylococcus aureus

Due to the increased emergence of drug-resistant bacteria, the declining efficiency of traditional antimicrobials has generated severe concerns in recent years. Subsequently, more interest in other antimicrobial agents from natural resources draws more attention as an alternative to conventional medications. This study investigated the bactericidal mechanism of monoterpene 1,8-cineol (eucalyptol), a major compound of various essential oils, against methicillin-resistant Staphylococcus aureus (MRSA). The antibacterial activity of 1,8-cineol was assessed by an MTT assay against clinical and reference MRSA strains. A cell membrane integrity test, followed by zeta potential (ZP) measurements, was performed to evaluate the disruption of the bacterial membrane integrity. Additionally, the cytotoxic effect of this molecule on MRSA bacteria was investigated by monitoring reactive oxygen species (ROS) generation, lipid peroxidation (MDA), and antioxidant enzyme activities (CAT and SOD). Regarding the anti-staphylococcal effect, the obtained results revealed the antibacterial efficacy of 1,8-cineol wherein the minimum inhibitory concentrations were equal to 7.23 mg/mL. Furthermore, it enhanced membrane permeability, with a 5.36-fold increase in nucleic acid and protein leakage as compared with untreated strains, along with the alteration of surface charge (ZP) in MRSA cells. The tested compound caused an increase in ROS generation reaching 17,462 FU and MDA production, reaching 9.56 μM/mg protein, in treated bacterial cells, along with a decrease in oxidative stress enzymes activities. Our findings suggest that 1,8-cineol has the ability to damage the membrane integrity and induce ROS-mediated oxidative stress in MRSA cells, leading to its antagonistic effect against this pathogen and consequently aiding in the reversal of antibiotic resistance.