Anti-Candida Targets and Cytotoxicity of Casuarinin Isolated from Plinia cauliflora Leaves in a Bioactivity-Guided Study
Abstract
:1. Introduction
2. Results and Discussion
2.1. Susceptibility Testing
C.a. (ATCC) | C.a. (SC5314) | C.k. | C.p. | C.t. | |||||||
---|---|---|---|---|---|---|---|---|---|---|---|
Samples | MIC | MFC | MIC | MFC | MIC | MFC | MIC | MFC | MIC | MFC | CC50 |
Extract | 156 | 625 | 156 | 156 | 19 | 39 | 78 | 156 | 312 | 1250 | 417 |
EAF | 625 | 1250 | NT | NT | 19 | 78 | 312 | 625 | 312 | 625 | 221 |
BF | 78 | 1250 | 156 | 156 | 19 | 39 | 19 | 19 | 156 | 1250 | 767 |
AF | 312 | >1250 | NT | NT | 39 | 78 | 39 | 78 | 312 | 1250 | 1500 |
F2 | 78 | 312 | 156 | 312 | 625 | >1250 | 156 | 1250 | 312 | >1250 | >400 |
casuarinin | 580 | 580 | 580 | >580 | 26 | 580 | 580 | 580 | 145 | 580 | >116 |
FCZ | 4 | 8 | 2 | 32 | 32 | 32 | 4 | 4 | 32 | 32 | NA |
Amph B | 0.25 | 0.25 | 1 | 8 | 0.50 | 0.50 | 0.50 | 0.50 | 0.50 | 2 | NA |
2.2. Ergosterol Levels
SAMPLES | C. albicans | C. krusei | C. parapsilosis | C. tropicalis |
---|---|---|---|---|
Control | 1.38 ± 0.32 | 1.13 ± 0.70 | 0.34 ± 0.30 | 1,94 ± 0.10 |
BF | 2.03 ± 0.77(+48) | 1.44 ± 0.58(+28) | 0.77 ± 0.70(+29) | 1,30 ± 0.86(−33) |
FCZ | 0.40 ± 0.23(−71) * | 0.68 ± 0.08(−40) | 0.05 ± 0.05(−84) * | 0(−100) * |
2.3. Reduction of P. Cauliflora Leaf Samples Antifungal Properties
2.3.1. Addition of Exogenous Ergosterol
2.3.2. Sorbitol Protection Assay
Ergosterol | Sorbitol | |||||||
---|---|---|---|---|---|---|---|---|
Samples | C.a. | C.k. | C.p. | C.t. | C.a. | C.k. | C.p. | C.t. |
Extract | 5000 | 39 | 312 | 625 | 2500 | 39 | 2500 | 156 |
EAF | 5000 | 39 | 1250 | 625 | 5000 | 39 | 2500 | 312 |
BF | 5000 | 39 | 1250 | 312 | 5000 | 39 | 1250 | 312 |
AF | 5000 | 78 | 2500 | 625 | 5000 | 78 | 2500 | 1250 |
F2 | 1250 | 78 | 1250 | 312 | 1250 | 78 | 625 | 312 |
FCZ | NA | NA | NA | NA | 8 | 64 | 4 | 64 |
Amph B | 2 | 16 | 16 | 4 | 0.25 | 1 | 1 | 1 |
2.4. Ultrastructural Analysis
2.5. Assessing C. albicans cell wall treated with F2
Control cells | Treated cells | |||
---|---|---|---|---|
Yeast | Hyphae | Yeast | Hyphae | |
Chitin (%) | 5.1 ± 1.8 | 21.8 ± 1.3 | 4.2 ± 1.4 | 22.1 ± 4.0 |
Glucan (%) | 66.3 ± 4.9 | 68.8 ± 0.6 | 73.4 ± 4.0 | 67.3 ± 2.0 |
Mannan (%) | 28.6 ± 5.6 | 9.4 ± 1.6 | 22.4 ± 5.3 | 10.6 ± 2.1 |
Covalently attached cell wall protein (µg/mL) | 109.4 ± 5.5 | 90.2 ± 5.0 | 114.5 ± 4.2 | 101.3 ± 2.0 * |
Non-covalently attached cell wall protein (µg/mL) | 226.5 ± 1.5 | 156.6 ± 0.5 | 227.6 ± 3.4 | 173.3 ± 0.1 |
3. Experimental
3.1. General Procedures
3.2. Plant Material
3.3. Extraction and Isolation
3.4. Fungal Strains, Cell Line and Culture Conditions
3.5. Antifungal Susceptibility Testing
3.6. Cytotoxicity Testing
3.7. Sterol Quantification Method
3.8. Ultrastructural Analysis by Transmission Electron Microscopy (TEM)
3.9. Cell Wall Polymer Quantification
3.10. Relative Cell Wall Porosity
3.11. Statistical Analysis
4. Conclusions
Acknowledgments
Conflicts of Interest
References
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Souza-Moreira, T.M.; Severi, J.A.; Lee, K.; Preechasuth, K.; Santos, E.; Gow, N.A.R.; Munro, C.A.; Vilegas, W.; Pietro, R.C.L.R. Anti-Candida Targets and Cytotoxicity of Casuarinin Isolated from Plinia cauliflora Leaves in a Bioactivity-Guided Study. Molecules 2013, 18, 8095-8108. https://doi.org/10.3390/molecules18078095
Souza-Moreira TM, Severi JA, Lee K, Preechasuth K, Santos E, Gow NAR, Munro CA, Vilegas W, Pietro RCLR. Anti-Candida Targets and Cytotoxicity of Casuarinin Isolated from Plinia cauliflora Leaves in a Bioactivity-Guided Study. Molecules. 2013; 18(7):8095-8108. https://doi.org/10.3390/molecules18078095
Chicago/Turabian StyleSouza-Moreira, Tatiana M., Juliana A. Severi, Keunsook Lee, Kanya Preechasuth, Emerson Santos, Neil A. R. Gow, Carol A. Munro, Wagner Vilegas, and Rosemeire C. L. R. Pietro. 2013. "Anti-Candida Targets and Cytotoxicity of Casuarinin Isolated from Plinia cauliflora Leaves in a Bioactivity-Guided Study" Molecules 18, no. 7: 8095-8108. https://doi.org/10.3390/molecules18078095