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Synergistic antibacterial effect between silybin and N,N′-dicyclohexylcarbodiimide in clinical Pseudomonas aeruginosa isolates

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Abstract

Silybin is a composition of the silymarin group as a hepatoprotective agent, and it exhibits various biological activities, including an antibacterial activity. In this study, the effects of a combination of silybin with N,N′-dicyclohexylcarbodiimide (DCCD) against clinical isolates of Pseudomonas aeruginosa were investigated. In the results of susceptibility assay, silybin showed more potent antibacterial activity in methicilin-resistant Staphylococcus aureus (MRSA) than in P. aeruginosa, but DCCD significantly increased the antibacterial activity of silybin in P. aeruginosa. The antibacterial activity of silybin was affected by the strong action of multidrug-resistant pumps rather than by a permeable disruption of lipopolysaccharide and silybin showed a remarkable synergistic activity in combination with some antibiotic agents against drug-resistant bacteria. Therefore, silybin has a potential as a combination therapeutic agent for treatment of infectious diseases by multidrug-resistant bacteria.

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References

  • Agarwal, C., R.P. Singh, S. Dhanalakshmi, A.K. Tyagi, M. Tecklenburg, R.A. Sclafani, and R. Agarwal. 2003. Silibinin upregulates the expression of cyclin-dependent kinase inhibitors and causes cell cycle arrest and apoptosis in human colon carcinoma HT-29 cells. Oncogene 22, 8271–8282.

    Article  PubMed  CAS  Google Scholar 

  • Cha, J.D., M.R. Jeong, S.I. Jeong, and K.Y. Lee. 2007. Antibacterial activity of sophoraflavanone G isolated from the roots of Sophora flavescens. J. Microbiol. Biotechnol. 17, 858–864.

    PubMed  CAS  Google Scholar 

  • Chen, C.J., J.E. Chin, K. Ueda, D.P. Clark, I. Pastan, M.M. Gottesman, and I.B. Roninson. 1986. Internal duplication and homology with bacterial transport proteins in the mdr1 (P-glycoprotein) gene from multidrug-resistant human cells. Cell 47, 381–389.

    Article  PubMed  CAS  Google Scholar 

  • Davidson, A.L. and J. Chen. 2004. ATP-binding cassette transporters in bacteria. Annu. Rev. Biochem. 73, 241–268.

    Article  PubMed  CAS  Google Scholar 

  • De Kievit, T.R., M.D. Parkins, R.J. Gillis, R. Srikumar, H. Ceri, K. Poole, B.H. Iglewski, and D.G. Storey. 2001. Multidrug efflux pumps: expression patterns and contribution to antibiotic resistance in Pseudomonas aeruginosa biofilms. Antimicrob. Agents Chemother. 45, 1761–1770.

    Article  PubMed  Google Scholar 

  • Gibbons, S. and E.E. Udo. 2000. The effect of reserpine, a modulator of multidrug efflux pumps, on the in vitro activity of tetracycline against clinical isolates of methicillin resistant Staphylococcus aureus (MRSA) possessing the tet(K) determinant. Phytother. Res. 14, 139–140.

    Article  PubMed  CAS  Google Scholar 

  • Goncalves, P.P., S.M. Meireles, P. Neves, and M.G. Vale. 1999. Synaptic vesicle Ca2+/H+ antiport: dependence on the proton electrochemical gradient. Mol. Brain Res. 71, 178–184.

    Article  PubMed  CAS  Google Scholar 

  • Gradelski, E., L. Valera, D. Bonner, and J. Fung-Tomc. 2001. Synergistic activities of gatifloxacin in combination with other antimicrobial agents against Pseudomonas aeruginosa and related species. Antimicrob. Agents Chemother. 45, 3220–3222.

    Article  PubMed  CAS  Google Scholar 

  • Gupta, O.P., S. Sing, S. Bani, N. Sharma, S. Malhotra, B.D. Gupta, S.K. Banerjee, and S.S. Handa. 2000. Anti-inflammatory and anti-arthritic activities of silymarin acting through inhibition of 5-lipoxygenase. Phytomedicine 7, 21–24.

    PubMed  CAS  Google Scholar 

  • Hancock, R.E. and P.G. Wong. 1984. Compounds which increase the permeability of the Pseudomonas aeruginosa outer membrane. Antimicrob. Agents Chemother. 26, 48–52.

    PubMed  CAS  Google Scholar 

  • Hooper, D.C. 2005. Efflux pumps and nosocomial antibiotic resistance: a primer for hospital epidemiologists. Clin. Infect. Dis. 40, 1811–1817.

    Article  PubMed  CAS  Google Scholar 

  • Irvin, R.T., T.J. Macalister, and J.W. Costerton. 1981. Tris(hydroxymethyl) aminomethane buffer modification of Escherichia coli outer membrane permeability. J. Bacteriol. 145, 1397–1403.

    PubMed  CAS  Google Scholar 

  • Jernaes, M.W. and H.B. Steen. 1994. Staining of Escherichia coli for flow cytometry: influx and efflux of ethidium bromide. Cytometry 17, 302–309.

    Article  PubMed  CAS  Google Scholar 

  • Jung, H.J., Y.B. Seu, and D.G. Lee. 2007. Candicidal action of resveratrol isolated from grapes on human pathogenic yeast C. albicans. J. Microbiol. Biotechnol. 17, 1324–1329.

    PubMed  CAS  Google Scholar 

  • Kren, V. and D. Walterova. 2005. Silybin and silymarin-new effects and applications. Biomed. Pap. Med. Fac. Univ. Palacky Olomouc. Czech Repub. 149, 29–41.

    PubMed  CAS  Google Scholar 

  • Leavis, H.L., R.J. Willems, J. Top, E. Spalburg, E.M. Mascini, A.C. Fluit, A. Hoepelman, A.J. De Neeling, and M.J. Bonten. 2003. Epidemic and nonepidemic multidrug-resistant Enterococcus faecium. Emerg. Infect. Dis. 9, 1108–1115.

    PubMed  Google Scholar 

  • Lee, E.W., M.N. Huda, T. Kuroda, T. Mizushima, and T. Tsuchiya. 2003a. EfrAB, an ABC multidrug efflux pump in Enterococcus faecalis. Antimicrob. Agents Chemother. 47, 3733–3738.

    Article  PubMed  CAS  Google Scholar 

  • Lee, D.G., H.K. Kim, Y. Park, S.C. Park, E.R. Woo, H.G. Jeong, and K.S. Hahm. 2003b. Gram-positive bacteria specific properties of silybin derived from Silybum marianum. Arch. Pharm. Res. 26, 597–600.

    PubMed  CAS  Google Scholar 

  • Leive, L. 1965. Actinomycin sensitivity in Escherichia coli produced by Edta. Biochem. Biophys. Res. Commun. 18, 13–17.

    Article  PubMed  CAS  Google Scholar 

  • Levy, S.B. 1992. Active efflux mechanisms for antimicrobial resistance. Antimicrob. Agents Chemother. 36, 695–703.

    PubMed  CAS  Google Scholar 

  • Lewis, K. 1994. Multidrug resistance pumps in bacteria: variations on a theme. Trends Biochem. Sci. 19, 119–123.

    Article  PubMed  CAS  Google Scholar 

  • Lewis, K. 2001. In search of natural substrates and inhibitors of MDR pumps. J. Mol. Microbiol. Biotechnol. 3, 247–254.

    PubMed  CAS  Google Scholar 

  • Lomovskaya, O. and K. Lewis. 1992. Emr, an Escherichia coli locus for multidrug resistance. Proc. Natl. Acad. Sci. USA 89, 8938–8942.

    Article  PubMed  CAS  Google Scholar 

  • Linnett, P.E. and R.B. Beechey. 1979. Inhibitors of the ATP synthethase system. Methods Enzymol. 55, 472–518.

    Article  PubMed  CAS  Google Scholar 

  • Mcmurry, L., R.E. Petrucci, Jr., and S.B. Levy. 1980. Active efflux of tetracycline encoded by four genetically different tetracycline resistance determinants in Escherichia coli. Proc. Natl. Acad. Sci. USA 77, 3974–3977.

    Article  PubMed  CAS  Google Scholar 

  • Nikaido, H. 2003. Molecular basis of bacterial outer membrane permeability revisited. Microbiol. Mol. Biol. Rev. 67, 593–656.

    Article  PubMed  CAS  Google Scholar 

  • Perez-Victoria, J.M., F.J. Perez-Victoria, G. Conseil, M. Maitrejean, G. Comte, D. Barron, A. Di Pietro, S. Castanys, and F. Gamarro. 2001. High-affinity binding of silybin derivatives to the nucleotide-binding domain of a Leishmania tropica P-glycoprotein-like transporter and chemosensitization of a multidrug-resistant parasite to daunomycin. Antimicrob. Agents Chemother. 45, 439–446.

    Article  PubMed  CAS  Google Scholar 

  • Renau, T.E., R. Leger, E.M. Flamme, J. Sangalang, M.W. She, R. Yen, C.L. Gannon, D. Griffith, S. Chamberland, O. Lomovskaya, S.J. Hecker, V.J. Lee, T. Ohta, and K. Nakayama. 1999. Inhibitors of efflux pumps in Pseudomonas aeruginosa potentiate the activity of the fluoroquinolone antibacterial levofloxacin. J. Med. Chem. 42, 4928–4931.

    Article  PubMed  CAS  Google Scholar 

  • Sharma, G., R.P. Singh, D.C. Chan, and R. Agarwal. 2003. Silibinin induces growth inhibition and apoptotic cell death in human lung carcinoma cells. Anticancer Res. 23, 2649–2655.

    PubMed  CAS  Google Scholar 

  • Singh, R.P. and R. Agarwal. 2004. Prostate cancer prevention by silibinin. Curr. Cancer Drug Targets 4, 1–11.

    Article  PubMed  CAS  Google Scholar 

  • Steinfels, E., C. Orelle, J.R. Fantino, O. Dalmas, J.L. Rigaud, F. Denizot, A. Di Pietro, and J.M. Jault. 2004. Characterization of YvcC (BmrA), a multidrug ABC transporter constitutively expressed in Bacillus subtilis. Biochemistry 43, 7491–7502.

    Article  PubMed  CAS  Google Scholar 

  • Stermitz, F.R., J. Tawara-Matsuda, P. Lorenz, P. Mueller, L. Zenewicz, and K. Lewis. 2000. 5′-Methoxyhydnocarpin-D and pheophorbide A: Berberis species components that potentiate berberine growth inhibition of resistant Staphylococcus aureus. J. Nat. Prod. 63, 1146–1149.

    Article  PubMed  CAS  Google Scholar 

  • Swallow, C.J., S. Grinstein, and O.D. Rotstein. 1990. A vacuolar type H(+)-ATPase regulates cytoplasmic pH in murine macrophages. J. Biol. Chem. 265, 7645–7654.

    PubMed  CAS  Google Scholar 

  • Tegos, G., F.R. Stermitz, O. Lomovskaya, and K. Lewis. 2002. Multidrug pump inhibitors uncover remarkable activity of plant antimicrobials. Antimicrob. Agents Chemother. 46, 3133–3141.

    Article  PubMed  CAS  Google Scholar 

  • Tyagi, A.K., C. Agarwal, D.C. Chan, and R. Agarwal. 2004. Synergistic anti-cancer effects of silibinin with conventional cytotoxic agents doxorubicin, cisplatin and carboplatin against human breast carcinoma MCF-7 and MDA-MB468 cells. Oncol. Rep. 11, 493–499.

    PubMed  CAS  Google Scholar 

  • Van Den Berg Van Saparoea, H.B., J. Lubelski, R. Van Merkerk, P.S. Mazurkiewicz, and A.J. Driessen. 2005. Proton motive force-dependent Hoechst 33342 transport by the ABC transporter LmrA of Lactococcus lactis. Biochemistry 44, 16931–16938.

    Article  PubMed  CAS  Google Scholar 

  • Van Der Heide, T. and B. Poolman. 2002. ABC transporters: one, two or four extracytoplasmic substrate-binding sites? EMBO Rep. 3, 938–943.

    Article  PubMed  Google Scholar 

  • Wellington, K. and B. Jarvis. 2001. Silymarin: a review of its clinical properties in the management of hepatic disorders. BioDrugs 15, 465–489.

    Article  PubMed  CAS  Google Scholar 

  • Yu, J.L., L. Grinius, and D.C. Hooper. 2002. NorA functions as a multidrug efflux protein in both cytoplasmic membrane vesicles and reconstituted proteoliposomes. J. Bacteriol. 184, 1370–1377.

    Article  PubMed  CAS  Google Scholar 

  • Zgurskaya, H.I. and H. Nikaido. 1999. Bypassing the periplasm: reconstitution of the AcrAB multidrug efflux pump of Escherichia coli. Proc. Natl. Acad. Sci. USA 96, 7190–7195.

    Article  PubMed  CAS  Google Scholar 

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  1. Department of Microbiology, College of Natural Sciences, Kyungpook National University, Daegu, 702-701, Republic of Korea

    Hyun Jun Jung & Dong Gun Lee

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  1. Hyun Jun Jung
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  2. Dong Gun Lee
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Correspondence to Dong Gun Lee.

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Jung, H.J., Lee, D.G. Synergistic antibacterial effect between silybin and N,N′-dicyclohexylcarbodiimide in clinical Pseudomonas aeruginosa isolates. J Microbiol. 46, 462–467 (2008). https://doi.org/10.1007/s12275-008-0138-7

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  • DOI: https://doi.org/10.1007/s12275-008-0138-7

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