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RND pump inhibition: in-silico and in-vitro study by Eugenol on clinical strain of E. coli and P. aeruginosa

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Abstract

Multidrug-resistant (MDR) gram-negative bacteria pose significant challenges to the public health. Various factors are involved in the development and spread of MDR strains, including the overuse and misuse of antibiotics, the lack of new antibiotics being developed, and etc. Efflux pump is one of the most important factors in the emergence of antibiotic resistance in bacteria. Aiming at the introduction of novel plant antibiotic, we investigated the effect of eugenol on the MexA and AcrA efflux pumps in Pseudomonas aeruginosa (P. aeruginosa) and Escherichia coli (E. coli). Molecular docking was performed using PachDock Server 1.3. The effect of eugenol on bacteria was determined by disk diffusion, minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC). A cartwheel test was also performed to evaluate efflux pump inhibition. Finally, the expression of the MexA and AcrA genes was examined by real-time PCR. The results of molecular docking showed that eugenol interacted with MexA and AcrA pumps at − 29.28 and − 28.59 Kcal.mol−1, respectively. The results of the antibiogram test indicated that the antibiotic resistance of the treated bacteria decreased significantly (p < 0.05). The results of the cartwheel test suggested the inhibition of efflux pump activity in P. aeruginosa and E. coli. Analysis of the genes by real-time PCR demonstrated that the expression of MexA and AcrA genes was significantly reduced, compared to untreated bacteria (p < 0.001). The findings suggest, among other things, that eugenol may make P. aeruginosa and E. coli more sensitive to antibiotics and that it could be used as an inhibitor to prevent bacteria from becoming resistant to antibiotics.

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Data availability

The datasets generated and/ or analyzed during the current study are avilable from the corresponding author on reasonable request.

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Acknowledgements

The authors thank Islamic Azad University for providing the infrastructure to conduct the present study.

Funding

This research received no specific funding from public, commercial, or nonprofit entities.

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Authors and Affiliations

  1. Department of Biology, Science and Research Branch, Islamic Azad University, Tehran, Iran

    Elham Etesami Ashtiani & Zohreh Gholizadeh Siahmazgi

  2. Department of Microbiology, Lahijan Branch, Islamic Azad University, Lahijan, Iran

    Mirsasan Mirpour

  3. Department of Genetics, Faculty of Biological Sciences, Tarbiat Modares University, Tehran, Iran

    Bahram Mohammad Soltani

Authors
  1. Elham Etesami Ashtiani
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  2. Zohreh Gholizadeh Siahmazgi
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  3. Mirsasan Mirpour
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  4. Bahram Mohammad Soltani
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EEA and ZGS prepared figures and tables. MM and BMS prepared manuscript. All authors reviewed the manuscript.

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Correspondence to Mirsasan Mirpour or Bahram Mohammad Soltani.

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Ashtiani, E.E., Gholizadeh Siahmazgi, Z., Mirpour, M. et al. RND pump inhibition: in-silico and in-vitro study by Eugenol on clinical strain of E. coli and P. aeruginosa. In Silico Pharmacol. 11, 22 (2023). https://doi.org/10.1007/s40203-023-00159-z

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