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Aucklandia lappa Causes Membrane Permeation of Candida albicans

  • Lee, Heung-Shick (Department of Biotechnology and Bioinformatics, Korea University) ;
  • Kim, Younhee (Department of Korean Medicine, Semyung University)
  • Received : 2020.09.25
  • Accepted : 2020.11.02
  • Published : 2020.12.28

Abstract

Candida albicans is a major fungal pathogen in humans. In our previous study, we reported that an ethanol extract from Aucklandia lappa weakens C. albicans cell wall by inhibiting synthesis or assembly of both (1,3)-β-D-glucan polymers and chitin. In the current study, we found that the extract is involved in permeabilization of C. albicans cell membranes. While uptake of ethidium bromide (EtBr) was 3.0% in control cells, it increased to 7.4% for 30 min in the presence of the A. lappa ethanol extract at its minimal inhibitory concentration (MIC), 0.78 mg/ml, compared to uptake by heat-killed cells. Besides, leakage of DNA and proteins was observed in A. lappa-treated C. albicans cells. The increased uptake of EtBr and leakage of cellular materials suggest that A. lappa ethanol extract induced functional changes in C. albicans cell membranes. Incorporation of diphenylhexatriene (DPH) into membranes in the A. lappa-treated C. albicans cells at its MIC decreased to 84.8%, after 60 min of incubation, compared with that of the controls, indicate that there was a change in membrane dynamics. Moreover, the anticandidal effect of the A. lappa ethanol extract was enhanced at a growth temperature of 40℃ compared to that at 35℃. The above data suggest that the antifungal activity of the A. lappa ethanol extract against C. albicans is associated with synergistic action of membrane permeabilization due to changes in membrane dynamics and cell wall damage caused by reduced formation of (1,3)-β-D-glucan and chitin.

Keywords

References

  1. Kullberg BJ, Arendrup MC. 2015. Invasive candidiasis. N. Engl. J. Med. 373: 1445-1456. https://doi.org/10.1056/NEJMra1315399
  2. Deorukhkar SC, Saini S, Mathew S. 2014. Non-albicans Candida infection: an emerging threat. Interdiscip. Perspect. Infect. Dis. 2014: 615958.
  3. Shapiro RS, Robbins N, Cowen LE. 2011. Regulatory circuitry governing fungal development, drug resistance, and disease. Microbiol. Mol. Biol. Rev. 75: 213-267. https://doi.org/10.1128/MMBR.00045-10
  4. Bondaryk M, Kurzatkowski W, Staniszewska M. 2013. Antifungal agents commonly used in the superficial and mucosal candidiasis treatment: mode of action and resistance development. Postepy Dermatol. Alergol. 30: 293-301.
  5. Odds FC, Brown AJ, Gow NA. 2003. Antifungal agents: mechanisms of action. Trends Microbiol. 11: 272-279. https://doi.org/10.1016/S0966-842X(03)00117-3
  6. Bossche HV, Koymans L, Moereels H. 1995. P450 inhibitors of use in medical treatment: focus on mechanisms of action. Pharmacol. Ther. 67: 79-100. https://doi.org/10.1016/0163-7258(95)00011-5
  7. Polak A, Scholer HJ. 1975. Mode of action of 5-fluorocytosine and mechanisms of resistance. Chemother 21: 113-130. https://doi.org/10.1159/000221854
  8. Kordalewska M, Perlin DS. 2019. Identification of drug resistant Candida auris. Front. Microbiol. 10: 1918.
  9. Guevara-Lora I, Bras G, Karkowska-Kuleta J, Gonzalez-Gonzalez M, Ceballos K, Sidlo W, et al. 2020. Plant-derived substances in the fight against infections caused by Candida species. Int. J. Mol. Sci. 21: 6131. https://doi.org/10.3390/ijms21176131
  10. Perumal Samy R, Gopalakrishnakone P. 2010. Therapeutic potential of plants as anti-microbials for drug discovery. Evid. Based Complement. Alternat. Med. 7: 283-294. https://doi.org/10.1093/ecam/nen036
  11. Lee HS, Kim Y. 2020. Aucklandia lappa causes cell wall damage in Candida albicans by reducing chitin and (1, 3)-β-D-glucan. J. Microbiol. Biotechnol. 30: 967-973. https://doi.org/10.4014/jmb.2002.02025
  12. Rodrigues L, Ramos J, Couto I, Amaral L, Viveiros M. 2011. Ethidium bromide transport across Mycobacterium smegmatis cell-wall: correlation with antibiotic resistance. BMC Microbiol. 11: 35. https://doi.org/10.1186/1471-2180-11-35
  13. Brasch J, Kreiselmaier I, Christophers E. 2003. Inhibition of dermatophytes by optical brighteners. Mycoses 46: 120-125. https://doi.org/10.1046/j.1439-0507.2003.00857.x
  14. Lee HS, Kim Y. 2017. Paeonia lactiflora inhibits cell wall synthesis and triggers membrane depolarization in Candida albicans. J. Microbiol. Biotechnol. 27: 395-404. https://doi.org/10.4014/jmb.1611.11064
  15. Chow J, Dionne HM, Prabhakar A, Mehrotra A, Somboonthum J, Gonzalez B, et al. 2019. Aggregate filamentous growth responses in yeast. mSphere 4: e00702-18.
  16. Kumar R, Saraswat D, Tati S, Edgerton M. 2015. Novel aggregation properties of Candida albicans secreted aspartyl proteinase Sap6 mediate virulence in oral candidiasis. Infect. Immun. 83: 2614-2026. https://doi.org/10.1128/IAI.00282-15
  17. LePecq JB, Paoletti C. 1967. A fluorescent complex between ethidium bromide and nucleic acids: physical-chemical characterization. J. Mol. Biol. 27: 87-106. https://doi.org/10.1016/0022-2836(67)90353-1
  18. Dive C, Watson JV, Workman P. 1990. Multiparametric analysis of cell membrane permeability by two colour flow cytometry with complementary fluorescent probes. Cytometry 11: 244-252. https://doi.org/10.1002/cyto.990110205
  19. Bradford MM. 1976. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal. Biochem. 72: 248-254. https://doi.org/10.1006/abio.1976.9999
  20. Repakova J, Capkova P, Holopainen JM, Vattulainen I. 2004. Distribution, orientation, and dynamics of DPH probes in DPPC bilayer. J. Phys. Chem. B. 108: 13438-13448. https://doi.org/10.1021/jp048381g
  21. Arino J, Ramos J, Sychrova H. 2010. Alkali metal cation transport and homeostasis in yeasts. Microbiol. Mol. Biol. Rev. 74: 95-120. https://doi.org/10.1128/MMBR.00042-09