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The efflux pump inhibitor tetrandrine exhibits synergism with fluconazole or voriconazole against Candida parapsilosis

  • Ya-Jing Zhao
  • Wei-Da Liu
  • Yong-Nian Shen
  • Dong-Mei Li
  • Kun-Ju Zhu
  • Hong ZhangEmail author
Original Article
  • 17 Downloads

Abstract

In the last two decades, with the wide use of azoles, antifungal resistance among Candida parapsilosis has considered a matter of concern worldwide. The aim of this study is to evaluate the antifungal potentials of tetrandrine (TET) alone and in combination with fluconazole (FLC)/voriconazole (VRC) against C. parapsilosis. Susceptibility tests were performed by microdilution method, checkerboard assay, time-kill test, spot assay. Subsequently, rhodamine 6G efflux test and the expressions of transporter related genes, namely CDR1 and MDR1 for C. parapsilosis were analyzed by qRT-PCR. The susceptibility test showed that TET presented strong synergism with FLC and VRC with fractional inhibitory concentration index (FICI) in a range of 0.094–0.562. The susceptibility results were also confirmed by spot assay and time-kill studies. With TET treatment, a vast quantity of rhodamine 6G could not be pumped out from the cells as considerably intracellular red fluorescence was accumulated. Meanwhile, the expressions of efflux-associated genes presented varying degrees of inhibition. These results indicated that TET was a decent antifungal synergist to promote the antifungal efficacy of FLC/VRC, and the underlying antifungal mechanism might be associated with the inhibition of efflux pump and the elevation of intracellular drug content.

Keywords

Tetrandrine (TET) Fluconazole (FLC) Voriconazole (VRC) Candida parapsilosis Synergism 

Notes

Author contributions

All the authors conceived the study, participated in its design and coordination and collected and managed the data. YZ drafted the manuscript, and all authors contributed substantially to its revision. All the authors read and approved the final manuscript.

Funding

This work was supported by the National Nature Science Foundation of China (Grant No. 81471995).

Compliance with ethical standards

Conflict of interest

All authors declare that they have no conflict of interest.

References

  1. 1.
    Pais P, Costa C, Pires C, Shimizu K, Chibana H, Teixeira MC (2016) Membrane proteome-wide response to the antifungal drug clotrimazole in Candida glabrata: role of the transcription factor CgPdr1 and the drug: H+ antiporters CgTpo1_1 and CgTpo1_2. Mol Cell Proteomics 15:57–72.  https://doi.org/10.1074/mcp.M114.045344 CrossRefGoogle Scholar
  2. 2.
    Marcos-Zambrano LJ, Escribano P, Sanchez C, Munoz P, Bouza E, Guinea J (2014) Antifungal resistance to fluconazole and echinocandins is not emerging in yeast isolates causing fungemia in a Spanish tertiary care center. Antimicrob Agents Chemother 58:4565–4572.  https://doi.org/10.1128/AAC.02670-14 CrossRefGoogle Scholar
  3. 3.
    Pammi M, Holland L, Butler G, Gacser A, Bliss JM (2013) Candida parapsilosis is a significant neonatal pathogen: a systematic review and meta-analysis. Pediatr Infect Dis J 32:e206–e216.  https://doi.org/10.1097/INF.0b013e3182863a1c CrossRefGoogle Scholar
  4. 4.
    Souza AC, Fuchs BB, Pinhati HM, Siqueira RA, Hagen F, Meis JF, Mylonakis E, Colombo AL (2015) Candida parapsilosis resistance to fluconazole: molecular mechanisms and in vivo impact in infected Galleria mellonella larvae. Antimicrob Agents Chemother 59:6581–6587.  https://doi.org/10.1128/AAC.01177-15 CrossRefGoogle Scholar
  5. 5.
    Berkow EL, Manigaba K, Parker JE, Barker KS, Kelly SL, Rogers PD (2015) Multidrug transporters and alterations in sterol biosynthesis contribute to azole antifungal resistance in Candida parapsilosis. Antimicrob Agents Chemother 59:5942–5950.  https://doi.org/10.1128/AAC.01358-15 CrossRefGoogle Scholar
  6. 6.
    Canton E, Peman J, Quindos G, Eraso E, Miranda-Zapico I, Alvarez M, Merino P, Campos-Herrero I, Marco F, de la Pedrosa EG, Yague G, Guna R, Rubio C, Miranda C, Pazos C, Velasco D, Group FS (2011) Prospective multicenter study of the epidemiology, molecular identification, and antifungal susceptibility of Candida parapsilosis, Candida orthopsilosis, and Candida metapsilosis isolated from patients with candidemia. Antimicrob Agents Chemother 55:5590–5596.  https://doi.org/10.1128/aac.00466-11 CrossRefGoogle Scholar
  7. 7.
    Falagas ME, Roussos N, Vardakas KZ (2010) Relative frequency of albicans and the various non-albicans Candida spp among candidemia isolates from inpatients in various parts of the world: a systematic review. Int J Infect Dis 14:e954–e966CrossRefGoogle Scholar
  8. 8.
    Zhang H, Wang K, Zhang G, Ho HI, Gao A (2010) Synergistic anti-candidal activity of tetrandrine on ketoconazole: an experimental study. Planta Med 76:53–61.  https://doi.org/10.1055/s-0029-1185973 CrossRefGoogle Scholar
  9. 9.
    Guo H, Xie SM, Li SX, Song YJ, Lv XL, Zhang H (2014) Synergistic mechanism for tetrandrine on fluconazole against Candida albicans through the mitochondrial aerobic respiratory metabolism pathway. J Med Microbiol 63:988–996.  https://doi.org/10.1099/jmm.0.073890-0 CrossRefGoogle Scholar
  10. 10.
    Li SX, Song YJ, Jiang L, Zhao YJ, Guo H, Li DM, Zhu KJ, Zhang H (2017) Synergistic effects of tetrandrine with posaconazole against Aspergillus fumigatus. Microb Drug Resist 23:674–681.  https://doi.org/10.1089/mdr.2016.0217 CrossRefGoogle Scholar
  11. 11.
    Li SX, Song YJ, Zhang LL, Shi JP, Ma ZL, Guo H, Dong HY, Li YM, Zhang H (2015) An in vitro and in vivo study on the synergistic effect and mechanism of itraconazole or voriconazole alone and in combination with tetrandrine against Aspergillus fumigatus. J Med Microbiol 64:1008–1020.  https://doi.org/10.1099/jmm.0.000120 CrossRefGoogle Scholar
  12. 12.
    Clinical and Laboratory Standard Institute (2008) Reference method for broth dilution antifungal susceptibility testing of yeasts; Approved Standard-Third Edition. CLSI document M27-A3. 2008. Clinical and Laboratory Standards Institute, Wayne, PAGoogle Scholar
  13. 13.
    Clinical and Laboratory Standard Institute (2012) Reference method for broth dilution antifungal susceptibility testing of yeasts; Fourth Informational Supplement M27-S4. Clinical and Laboratory Standards Institute, Wayne, PAGoogle Scholar
  14. 14.
    Sun S, Li Y, Guo Q, Shi C, Yu J, Ma L (2008) In vitro interactions between tacrolimus and azoles against Candida albicans determined by different methods. Antimicrob Agents Chemother 52:409–417.  https://doi.org/10.1128/AAC.01070-07 CrossRefGoogle Scholar
  15. 15.
    Shao J, Shi G, Wang T, Wu D, Wang C (2016) Antiproliferation of berberine in combination with fluconazole from the perspectives of reactive oxygen species, ergosterol and drug efflux in a fluconazole-resistant Candida tropicalis isolate. Front Microbiol 7:1516.  https://doi.org/10.3389/fmicb.2016.01516 CrossRefGoogle Scholar
  16. 16.
    Wang T, Shao J, Da W, Li Q, Shi G, Wu D, Wang C (2018) Strong synergism of palmatine and fluconazole/itraconazole against planktonic and biofilm cells of Candida species and efflux-associated antifungal mechanism. Front Microbiol 9:2892.  https://doi.org/10.3389/fmicb.2018.02892 CrossRefGoogle Scholar
  17. 17.
    Singh P, Kaur J, Yadav B, Komath SS (2010) Targeting efflux pumps-in vitro investigations with acridone derivatives and identification of a lead molecule for MDR modulation. Bioorg Med Chem 18:4212–4223.  https://doi.org/10.1016/j.bmc.2010.05.003 CrossRefGoogle Scholar
  18. 18.
    Maesaki S, Marichal P, Vanden Bossche H, Sanglard D, Kohno S (1999) Rhodamine 6G efflux for the detection of CDR1-overexpressing azole-resistant Candida albicans strains. J Antimicrob Chemother 44:27–31.  https://doi.org/10.1093/jac/44.1.27 CrossRefGoogle Scholar
  19. 19.
    Fakhim H, Chowdhary A, Prakash A, Vaezi A, Dannaoui E, Meis JF, Badali H (2017) In vitro interactions of echinocandins with triazoles against multidrug-resistant Candida auris. Antimicrob Agents Chemother 61:e01056-17.  https://doi.org/10.1128/AAC.01056-17 CrossRefGoogle Scholar
  20. 20.
    Ahmad A, Khan A, Manzoor N (2013) Reversal of efflux mediated antifungal resistance underlies synergistic activity of two monoterpenes with fluconazole. Eur J Pharm Sci 48:80–86.  https://doi.org/10.1016/j.ejps.2012.09.016 CrossRefGoogle Scholar
  21. 21.
    Jia W, Zhang H, Li C, Li G, Liu X, Wei J (2016) The calcineruin inhibitor cyclosporine a synergistically enhances the susceptibility of Candida albicans biofilms to fluconazole by multiple mechanisms. BMC Microbiol 16:113.  https://doi.org/10.1186/s12866-016-0728-1 CrossRefGoogle Scholar

Copyright information

© Springer Nature B.V. 2019

Authors and Affiliations

  • Ya-Jing Zhao
    • 1
    • 2
  • Wei-Da Liu
    • 3
  • Yong-Nian Shen
    • 3
  • Dong-Mei Li
    • 4
  • Kun-Ju Zhu
    • 1
    • 2
  • Hong Zhang
    • 1
    • 2
    Email author
  1. 1.The First Affiliated Hospital of Jinan UniversityGuangzhouChina
  2. 2.Institute of MycologyJinan UniversityGuangzhouChina
  3. 3.Department of Mycology, Institute of DermatologyChinese Academy of Medical Science and Peking Union Medical CollegeNanjingChina
  4. 4.Department of Microbiology and ImmunologyGeorgetown University Medical CenterWashingtonUSA

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