Enhancement of CREC sensitivity to fosfomycin by baicalein through increasing outer membrane permeability

Background:To confirm whether baicalein improves the sensitivity of carbapenem-resistant Escherichia coli(CREC)to fosfomycin by increasing the permeability of bacterial outer membrane in vitro experiments.Methods:The clinically isolated CREC strains were amplified and then divided into three groups including baicalein monotherapy groups,fosfomycin monotherapy groups,and baicalein plus fosfomycin groups,and their minimum inhibitory concentrations(MICs)measurement and interpretation were performed according to CLSI interpretive criteria.To determine bacterial permeability after contact with baicalein,CREC were incubated with fluorescein isothiocyanate(FITC)after pretreatment with blank control without baicalein,with 0.25 MIC of baicalein,and with 0.125 MIC of baicalein,followed by observation of the intrabacterial fluorescence intensity of FITC.In addition,CREC were pretreated with 0.125 MIC of baicalein and with blank control without baicalein followed by measurement of alkaline phosphatase(AKP)leak to determine the change of bacterial permeability.Results:The MIC range in baicalein monotherapy groups was from 128 mg/L to 256 mg/L,and the MIC range in fosfomycin monotherapy groups was from 16 mg/L to 1,024 mg/L,but the MIC range in both combination therapy groups was reduced to 4 mg/L to 64 mg/L.The combination use reduced the MIC of each therapy by 75%-96.88%in all strains,and the fractional inhibitory concentration index(FICI)values less than or equal to 0.5.In the permeability assay,no permeabilization of FITC was observed in the blank groups without baicalein,but the intrabacterial FITC aggregation was observed in the groups of pretreatment with 0.25 MIC of baicalein or 0.25 MIC of baicalein.In the AKP leak assay,the AKP leak was more severe at the groups of coincubation with 0.25 MIC of baicalein than those blank groups without baicalein within the first 6 hours.Conclusion:Our study suggests that baicalein may synergistically enhance the antibacterial effect of fosfomycin by increasing the permeability of CREC outer membrane.