Mechanism of Protodioscin Against Carbapenem-resistant Raoultella Ornithinolytica B1645-1B Carrying NDM-1 in Vitro
Objective To investigate the antibacterial effect of protodioscin(PD)against carbapenem-resistant Raoultella ornithinolytica B1645-1B carrying NDM-1 in vitro and its mechanism.Methods A broth microdilution method was used to determine the minimum inhibitory concentration(MIC)of PD against strain B1645-1B.The MIC of PD combined with imi-penem(IMP)against the strain was determined by the checkerboard method,and the fractional inhibitory concentration(FIC)index was calculated.The mRNA expression of NDM-1 in strain B1645-1B after treatment with PD was detected by quantitative real-time PCR(qRT-PCR).The Molecular Operating Environment(MOE)was used to predict the molecular docking between PD and NDM-1 protein.The protein expression of NDM-1 after treatment with PD and purification was determined by sodium dodecyl sulfate-polyacrylamide gel electrophoresis(SDS-PAGE).Results The MIC of PD against strain B1645-1B was 10 000 μg/mL,and PD combined with IMP had an FIC of 0.75,indicating synergy.After treatment with PD,the mRNA synthesis and enzyme activity of NDM-1 were inhibited.Conclusion PD was a promising novel and effective synergistic antimicrobial agent and NDM-1 enzyme inhibitor.
ProtodioscinRaoultella ornithinolyticaNDM-1ImipenemAntibacterial experiment in vitro
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