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. 2022 Dec 19;11(12):1842.
doi: 10.3390/antibiotics11121842.

In Silico and In Vitro Analysis of Sulforaphane Anti- Candida Activity

Bruna L R Silva  1 , Gisele Simão  2   3 , Carmem D L Campos  4 , Cinara R A V Monteiro  4 , Laryssa R Bueno  2   3 , Gabriel B Ortis  2   3 , Saulo J F Mendes  5 , Israel Viegas Moreira  4 , Daniele Maria-Ferreira  2   3 , Eduardo M Sousa  1   5 , Flávia C B Vidal  6 , Cristina de Andrade Monteiro  7 , Valério Monteiro-Neto  1   4 , Elizabeth S Fernandes  2   3
Affiliations

In Silico and In Vitro Analysis of Sulforaphane Anti- Candida Activity

Bruna L R Silva et al. Antibiotics (Basel). .

Abstract

Oropharyngeal candidiasis/candidosis is a common and recurrent opportunistic fungal infection. Fluconazole (FLZ), one of the most used and effective antifungal agents, has been associated with a rise of resistant Candida species in immunocompromised patients undergoing prophylactic therapy. Sulforaphane (SFN), a compound from cruciferous vegetables, is an antimicrobial with yet controversial activities and mechanisms on fungi. Herein, the in silico and antifungal activities of SFN against C. albicans were investigated. In silico analyzes for the prediction of the biological activities and oral bioavailability of SFN, its possible toxicity and pharmacokinetic parameters, as well as the estimates of its gastrointestinal absorption, permeability to the blood-brain barrier and skin, and similarities to drugs, were performed by using different software. SFN in vitro anti-Candida activities alone and in combination with fluconazole (FLZ) were determined by the broth microdilution method and the checkerboard, biofilm and hyphae formation tests. Amongst the identified probable biological activities of SFN, nine indicated an antimicrobial potential. SFN was predicted to be highly absorbable by the gastrointestinal tract, to present good oral availability, and not to be irritant and/or hepatotoxic. SFN presented antifungal activity against C. albicans and prevented both biofilm and hyphae formation by this microorganism. SFN was additive/synergistic to FLZ. Overall, the data highlights the anti-Candida activity of SFN and its potential to be used as an adjuvant therapy to FLZ in clinical settings.

Keywords: Candida albicans; antifungal activity; in silico analysis; sulforaphane.

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Conflict of interest statement

The authors declare no conflict of interest.

Figures

Figure 1
Figure 1
Effects of sulforaphane (SFN) and fluconazole (FLZ) on hyphae formation by C. albicans ATCC 90028. (a) Representative images of hyphae growth by C. albicans ATCC 90028 incubated with either FLZ or SFN at MIC/2 and MIC/4. The same panel represents the combined effects of SFN (MIC/64 to MIC/256) and FLZ (MIC/2 and MIC/4). (b) Percentage of hyphae per 100 cells of C. albicans ATCC 90028 and (c) percentage (%) of inhibition of C. albicans ATCC90028 hyphae formation. The control consisted of inoculum plus phosphate-buffered saline (PBS) containing 10% fetal bovine serum (FBS). * p < 0.05 differs from the control group.
Figure 2
Figure 2
Effects of sulforaphane (SFN) and fluconazole (FLZ) on hyphae formation by C. albicans oral isolate 40 HIV+. (a) Representative images of hyphae growth by Oral 40 HIV+ incubated with either FLZ or SFN at MIC/2 and MIC/4. The same panel represents the combined effects of SFN (MIC/64 to MIC/256) and FLZ (MIC/4 and MIC/8). (b) Percentage of hyphae per 100 cells of Oral 40 HIV+ and (c) percentage (%) of inhibition of Oral 40 HIV+ hyphae formation. The control consisted of inoculum plus phosphate-buffered saline (PBS) containing 10% fetal bovine serum (FBS). * p < 0.05 differs from the control group.
Figure 3
Figure 3
Effects of sulforaphane (SFN) and fluconazole (FLZ) on biofilm formation by C. albicans spp. SFN and FLZ were tested at MIC/2 and MIC/4 against (a) C. albicans ATCC 90028 and (b) C. albicans clinical isolate (Oral 40 HIV+). The combined effects of SFN (MIC/64 to MIC/256) with FLZ (MIC/2–MIC/8) against (c) C. albicans ATCC 90028 and (d) C. albicans clinical isolate (Oral 40 HIV+) were investigated over 24 h. The control consisted of inoculum plus culture medium. Data were obtained from three independent experiments and are expressed as Log10 CFU/mL.* p < 0.05 differs from the control group.
Figure 4
Figure 4
Effects of sulforaphane (SFN) and fluconazole (FLZ) on mRNA expression of hyphae growth- and biofilm formation-related genes by C. albicans spp. SFN and FLZ were tested at MIC/2 and MIC/4 against C. albicans ATCC 90028 and the oral isolate 40 HIV+. The relative expressions of ALS1 (a,d), EFG1 (b,e) and RAS1 (c,f) normalized to 18S rRNA were quantified 24 h following incubation with either FLZ or SFN. The control consisted of inoculum plus culture medium. Data were obtained from three independent experiments and are depicted as relative expression of mRNA. * p < 0.05 differs from the control group.
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