Volume 28, Issue 6 (1-2026)
J Arak Uni Med Sci 2026, 28(6): 420-431 |
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Khodavandi A, Alizadeh F, Rastgo P. Efficacy of Thymus vulgaris and Trigonella foenum-graecum Extracts Alone and in Combination in Candida albicans Hyphae Model and in Downregulation of Secreted Aspartyl Proteinase 1 Gene Expression. J Arak Uni Med Sci 2026; 28 (6) :420-431
URL: http://jams.arakmu.ac.ir/article-1-7963-en.html
URL: http://jams.arakmu.ac.ir/article-1-7963-en.html
1- Department of Microbiology, Gac. C., Islamic Azad University, Gachsaran, Iran
2- Department of Microbiology, Gac. C., Islamic Azad University, Gachsaran, Iran ,fahimeh.alizadeh@iau.ac.ir
3- Department of Microbiology, Yas. C., Islamic Azad University, Yasuj, Iran
2- Department of Microbiology, Gac. C., Islamic Azad University, Gachsaran, Iran ,
3- Department of Microbiology, Yas. C., Islamic Azad University, Yasuj, Iran
Abstract: (832 Views)
Introduction: The emergence of antifungal resistance in Candida albicans diseases poses a threat to global public health. New treatments are needed to target C. albicans and its ability to produce hyphae. The aim of this study was to investigate the potential synergistic effects and antifungal properties of Thymus vulgaris and Trigonella foenum-graecum extracts alone and in combination on C. albicans.
Methods: In this experimental study, extracts of Thymus vulgaris and Trigonella foenum-graecum were prepared using hot water (60 °C) and Soxhlet extraction with methanol (10%). Yeast susceptibility testing was performed according to the Clinical and Laboratory Standards Institute disk diffusion and broth microdilution guidelines. The hyphal model was created in the presence of alcoholic extracts of Thymus vulgaris and Trigonella foenum-graecum alone and in combination. Crystal violet staining, microscopic observation and gene expression analysis were used to evaluate the inhibition of hyphal growth.
Results: The results showed that 90% of the C. albicans isolates were resistant to fluconazole. Aqueous and alcoholic extracts of Thymus vulgaris in combination with Trigonella foenum-graecum showed synergistic, partially synergistic and additive effects. Alcoholic extracts of Thymus vulgaris with Trigonella foenum-graecum alone and in combination have anti-hyphae activity by reducing the percentage of hyphae, reducing the number of planktonic cells and the transition of planktonic cells to hyphae, and down-regulating the Secreted Aspartyl Proteinase 1 (SAP1) gene.
Conclusions: Taken together, these results indicate that extracts of Thymus vulgaris alone and in combination with Trigonella foenum-graecum may offer a potent alternative strategy to combat resistant C. albicans infections and their ability to reduce hyphae formation. Additionally, the SAP1 gene could be a likely target in the synergistic interaction of alcoholic extracts of Thymus vulgaris in combination with Trigonella foenum-graecum against the C. albicans hyphae model.
Methods: In this experimental study, extracts of Thymus vulgaris and Trigonella foenum-graecum were prepared using hot water (60 °C) and Soxhlet extraction with methanol (10%). Yeast susceptibility testing was performed according to the Clinical and Laboratory Standards Institute disk diffusion and broth microdilution guidelines. The hyphal model was created in the presence of alcoholic extracts of Thymus vulgaris and Trigonella foenum-graecum alone and in combination. Crystal violet staining, microscopic observation and gene expression analysis were used to evaluate the inhibition of hyphal growth.
Results: The results showed that 90% of the C. albicans isolates were resistant to fluconazole. Aqueous and alcoholic extracts of Thymus vulgaris in combination with Trigonella foenum-graecum showed synergistic, partially synergistic and additive effects. Alcoholic extracts of Thymus vulgaris with Trigonella foenum-graecum alone and in combination have anti-hyphae activity by reducing the percentage of hyphae, reducing the number of planktonic cells and the transition of planktonic cells to hyphae, and down-regulating the Secreted Aspartyl Proteinase 1 (SAP1) gene.
Conclusions: Taken together, these results indicate that extracts of Thymus vulgaris alone and in combination with Trigonella foenum-graecum may offer a potent alternative strategy to combat resistant C. albicans infections and their ability to reduce hyphae formation. Additionally, the SAP1 gene could be a likely target in the synergistic interaction of alcoholic extracts of Thymus vulgaris in combination with Trigonella foenum-graecum against the C. albicans hyphae model.
Keywords: Candida albicans, Thymus vulgaris and Trigonella foenum-graecum in combination, SAP1 gene expression
Type of Study: Original Atricle |
Subject:
Basic Sciences
Received: 2025/02/16 | Accepted: 2025/05/18
Received: 2025/02/16 | Accepted: 2025/05/18
References
1. Miranda S, Lassnig C, Schmidhofer K, et al. Lack of TYK2 signaling enhances host resistance to Candida albicans skin infection. Nat Commun. 2024;15(1):10493. pmid: 39622833 doi: 10.1038/s41467-024-54888-6
2. Du Toit A. Hyphae promote Candida albicans fitness and commensalism in the gut. Nat Rev Microbiol. 2024 22( 5) 258. pmid: 38472546 doi: 10.1038/s41579-024-01040-2
3. Abdulghani M, Zore G. Clinical significance, molecular formation, and natural antibiofilm agents of Candida albicans. In: Advances in Antifungal Drug Development: Natural Products with Antifungal Potential. Singapore: Springer Nature Singapore; 2024:251-291.
4. Bu QR, Bao MY, Yang Y, Wang TM, Wang CZ. Targeting virulence factors of Candida albicans with natural products. Foods. 2022;11(19):2951. pmid: 36230026 doi: 10.3390/foods11192951
5. Bras G, Satala D, Juszczak M, et al. Secreted aspartic proteinases: key factors in Candida infections and host-pathogen interactions. Int J Mol Sci. 2024;25(9):4775. pmid: 38731993 doi: 10.3390/ijms25094775
6. Katsipoulaki M, Stappers MHT, Malavia-Jones D, Brunke S, Hube B, Gow NAR. Candida albicans and Candida glabrata: global priority pathogens. Microbiol Mol Biol Rev. 2024;88(2):e0002123. pmid: 38832801 doi: 10.1128/mmbr.00021-23
7. Varshan GA, Namasivayam SKR. A critical review on sustainable formulation of anti-quorum sensing compounds using nanotechnology principles against Candida albicans. BioNanoSci. 2025;15(1):161. doi: 10.1007/s12668-024-01685-6
8. Barbosa PF, Gonçalves DS, Ramos LS, et al. Saps1-3 antigens in Candida albicans: differential modulation following exposure to soluble proteins, mammalian cells, and infection in mice. Infect Dis Rep. 2024;16(4):572-586. pmid: 39051243 doi: 10.3390/idr16040043
9. Silva NC, Nery JM, Dias AL. Aspartic proteinases of Candida spp.: role in pathogenicity and antifungal resistance. Mycoses. 2014;57(1):1-11. pmid: 23735296 doi: 10.1111/myc.12095
10. Khodavandi A, Alizadeh F, Khezrian F. Inhibition of Candida albicans yeast–hyphal transition by combination of fluconazole with amphotericin B. Physiol Pharmacol. 2018;22(3),195-204.
11. Hosseini SS, Yadegari MH, Rajabibazl M, Ghaemi EA. Inhibitory effects of carvacrol on the expression of secreted aspartyl proteinases 1-3 in fluconazole-resistant Candida albicans isolates. Iran J Microbiol. 2016;8(6):401-409. pmid: 28491252
12. Wirtu SF, Ramaswamy K, Maitra R, Chopra S, Mishra AK, Jule LT. Isolation, characterization and antimicrobial activity study of Thymus vulgaris. Sci Rep. 2024;14(1):21573. pmid: 39284874 doi: 10.1038/s41598-024-71012-2
13. Katnoria H, Kaushal S, Hunjan MS, Kaur V, Sharma P, Jangra R. Chemical characterization and antifungal potential of Trigonella foenum-graecum L. followed by molecular docking studies. J Essent Oil Bear Pl. 2024;27(1):57-72. doi: 10.1080/0972060X.2024.2302110
14. Khodavandi A, Alizadeh F. Hakimizadeh S. Inhibitory effect of aqueous and ethanolic extracts of chavill leaf and stem on aspartyl proteinase secreted from Candida albicans. J North Khorasan Univ Med Sci. 2018;9(4):91-99. [Persian].
15. CLSI (Clinical and Laboratory Standards Institute). Reference method for broth dilution antifungal susceptibility testing of yeasts. Approved standard M27-A3, third ed. Clinical and Laboratory Standards Institute, 2008; Wayne, PA, USA.
16. CLSI (Clinical and Laboratory Standards Institute). Reference method for broth dilution antifungal susceptibility testing of yeasts; fourth informational supplement, M27-S4. Clinical and Laboratory Standards Institute, 2012; Wayne, PA, USA.
17. Khodavandi A, Alizadeh F, Marashi N. Antibiofilm Activity of fluconazole/terbinafine combination in Candida albicans HWP1 gene expression. J Arak Uni Med Sci. 2018;20(11):22-33.
18. Khodavandi A, Harmal NS, Alizadeh F et al. Comparison between allicin and fluconazole in Candida albicans biofilm inhibition and in suppression of HWP1 gene expression. Phytomedicine. 2011;19(1):56-63. pmid: 21924600 doi: 10.1016/j.phymed.2011.08.060
19. Norouzi N, Alizadeh F, Khodavandi A, Jahangiri M. Antifungal activity of menthol alone and in combination on growth inhibition and biofilm formation of Candida albicans. J Herb Med. 2021;29:100495. doi: 10.1016/j.hermed.2021.100495
20. Esfahani MB, Khodavandi A, Alizadeh F, Bahador N. Possible molecular targeting of biofilm-associated genes by nano-Ag in Candida albicans. Appl Biochem Biotechnol. 2024;196(7):4205-4233. pmid: 37922031 doi: 10.1007/s12010-023-04758-6
21. Khodavandi A, Alizadeh F, Abrahe Z. Comparison of ERG11 gene expression profiles of Candida albicans treated with Thymus vulgaris extracts alone and in combination with Mentha spicata. J Microbial Biol. 2018;7(25):87-99. [Persian].
22. Khodavandi A, Alizadeh F, Shahinipor M. Relative quantitation of hyphae-specific gene HWP1 expression in inhibition of Candida albicans biofilm. J Microb World. 2016; 9(1):22-33. [Persian].
23. Khan S, Imran M, Imran M, Pindari N. Antimicrobial activity of various ethanolic plant extracts against pathogenic multi drug resistant Candida spp. Bioinformation. 2017;13(3):67-72. pmid: 28584446 doi: 10.6026/97320630013067
24. Sadoon Abd H, Al Haidar AHM. Comparison of antifungal activity of thymus vulgaris essential oil and triple antibiotic paste against Candida albicans isolated from root canal (in vitro study). F1000Research. 2024;13:381.
25. Sindhusha VB, Rajasekar A. Preparation and evaluation of antimicrobial property and anti-inflammatory activity of fenugreek gel against oral microbes: an invitro study. Cureus. 2023;15(10):e47659. pmid: 38022270 doi: 10.7759/cureus.47659
26. Maxwell SY, Gadallah MAEA. Comparison between the anti-Candida activity of fenugreek and ginger rhizome extracts and their synergism with fluconazole and nystatin. Egyptian J Med Microbio. 2023;32(2):59-64. doi: 10.21608/ejmm.2023.285225
27. Yadav SK, Jain GK, Mazumder A, Khar RK. Antimicrobial activity of a novel polyherbal combination for the treatment of vaginal infection. J Adv Pharm Technol Res. 2019;10(4):190-194. pmid: 31742120 doi: 10.4103/japtr.JAPTR_3_19
28. Sharma K, Parmanu PK, Sharma M. Mechanisms of antifungal resistance and developments in alternative strategies to combat Candida albicans infection. Arch Microbiol. 2024;206(3):95. pmid: 38349529 doi: 10.1007/s00203-023-03824-1
29. Zobi C, Algul O. The significance of mono‐and dual‐effective agents in the development of new antifungal strategies. Chem Biol Drug Des. 2025;105(1):e70045. doi: 10.1111/cbdd.70045
30. Yue D, Zheng D, Bai Y, Yang L, Yong J, Li Y. Insights into the anti-Candida albicans properties of natural phytochemicals: An in vitro and in vivo investigation. Phytother Res. 2024;38(5):2518-2538. pmid: 38450815 doi: 10.1002/ptr.8148
31. Long N, Li F. Antifungal mechanism of natural products derived from plants: a review. Nat Prod Commun. 2024;19(8):1934578X241271747. doi: 10.1177/1934578X241271747
32. Ben Miri Y. Essential oils: chemical composition and diverse biological activities: a comprehensive review. Nat Prod Commun. 2025;20(1):1934578X241311790. doi: 10.56027/JOASD282023
33. Sultan S, Zoofeen U, Shah I, et al. Enhanced inflammatory and oxidative response mitigation by acetyl-L-carnitine in a rat model of pelvic inflammatory disease. Naunyn Schmiedebergs Arch Pharmacol. 2025. pmid: 39912899 doi: 10.1007/s00210-025-03858-w
34. Chouhan S, Sharma K, Guleria S. Antimicrobial activity of some essential oils-present status and future perspectives. Medicines (Basel). 2017;4(3):58. pmid: 28930272 doi: 10.3390/medicines4030058
35. Ivanova S, Gvozdeva Y, Staynova R, et al. Essential oils–a review of the natural evolution of applications and some future perspectives. Pharmacia. 2025;72:1-12. doi: 10.3897/pharmacia.72.e140059
36. Essid R, Gharbi D, Abid G, et al. Combined effect of Thymus capitatus and Cinnamomum verum essential oils with conventional drugs against Candida albicans biofilm formation and elucidation of the molecular mechanism of action. Ind Crops Prod. 2019;140:111720.
37. Benzaid C, Belmadani A, Djeribi R, Rouabhia M. The effects of Mentha × piperita essential oil on C. albicans growth, transition, biofilm formation, and the expression of secreted aspartyl proteinases genes. Antibiotics (Basel). 2019;8(1):10. pmid: 30704020 doi: 10.3390/antibiotics8010010
38. Shaygan S, Khodavandi A. Inhibitory effect of menthol on expression of aspartyl proteinase 1 in fluconazole-resistant Candida albicans. J Herbmed Pharmacol. 2019;8(1):35-40. doi: 10.15171/jhp.2019.06
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