Inhibitory Effects and Mechanism of Action of Elsinochrome A on Candida albicans and Its Biofilm
Abstract
:1. Introduction
2. Materials and Methods
2.1. Strains and Growth Conditions
2.2. Antifungal Susceptibility Testing
2.3. Effect of PACT on the Growth Kinetics of C. albicans
2.4. Effect of PACT on Biofilm Formation
2.5. Time-Dependent Photoinactivation Effect
2.6. Biofilm Initial Cell Attachment Assay
2.6.1. XTT Assay for the Effect of EA on the Adhesion of C. albicans
2.6.2. Adhesion-Related Gene Transcription Analysis by qRT-PCR Assays
2.7. Biofilm Eradication Assay
2.8. Effect of PACT on Cell Permeability
2.9. Effect of PACT on Relative Electric Conductivity of Cell Membrane
2.10. Effect of PACT on Plasma Membrane Dynamics
2.11. Effect of PACT on ROS Production
2.11.1. Determination of ROS Production
2.11.2. Oxidative-Stress-Related Genes Transcription Analysis by qRT-PCR Assays
2.12. Mitochondrial Membrane Potential
2.13. Detection of DNA Fragmentation
Primers | Sequence | References |
---|---|---|
MP65-F | TCAACACTGAACCACCTC | [31] |
MP65-R | ATACCTTTAGCACCACCA | [31] |
ALS1-F | CATCATTGACTCAGTTGT | [32] |
ALS1-R | CAGTGGAAGTAGATTGTG | [32] |
CAP1-F | AGTCAATTCAATGTTCAAG | [32] |
CAP1-R | AATGGTAATGTCCTCAAG | [32] |
CAT1-F | GACTGCTTACATTCAAAC | [32] |
CAT1-R | AACTTACCAAATCTTCTCA | [32] |
SOD1-F | TTGAACAAGAATCCGAATCC | [33] |
SOD1-R | AGCCAATGACACCACAAGCAG | [33] |
ACT1-F | TTGATTTGGCTGGTAGAG | [34] |
ACT1-R | ATGGCAGAAGATTGAGAA | [34] |
2.14. Statistical Analysis
3. Results
3.1. EA-Mediated PACT Decreases C. albicans Survival
3.2. EA-Mediated PACT Inhibits C. albicans Biofilm Formation
3.3. EA-Mediated PACT Has Anti-Biofilm Activity toward Two Phases of C. albicans Biofilm Maturation
3.4. Effect of EA-Mediated PACT on Cell Membrane
3.4.1. Permeability of Cell Membrane
3.4.2. Integrity of Cell Membrane
3.5. EA-Mediated PACT Induces ROS Production
3.6. EA-Mediated PACT Decreases Mitochondrial Membrane Potential and Induces Nuclear Fragmentation
4. Discussion
4.1. EA-Mediated PACT Effect on C. albicans
4.2. EA-Mediated PACT Effect on C. albicans Biofilm
4.3. Mechanism of Action of EA-Mediated PACT
5. Conclusions
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Data Availability Statement
Acknowledgments
Conflicts of Interest
References
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Irradiation | Time | Fluconazole | |||||
---|---|---|---|---|---|---|---|
0 min | 5 min | 10 min | 20 min | 40 min | 60 min | ||
SMIC80 (μg/Ml) | 256–512 | 256–512 | 128–256 | 64–128 | 16–32 | 16 | >1024 |
MIC (μg/mL) | 128 | 16 | 8 | 4 | 2 | 1 | 0.5 |
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Pan, L.; Yao, Y.; Zheng, H.; Yan, S.; Chen, S. Inhibitory Effects and Mechanism of Action of Elsinochrome A on Candida albicans and Its Biofilm. J. Fungi 2022, 8, 841. https://doi.org/10.3390/jof8080841
Pan L, Yao Y, Zheng H, Yan S, Chen S. Inhibitory Effects and Mechanism of Action of Elsinochrome A on Candida albicans and Its Biofilm. Journal of Fungi. 2022; 8(8):841. https://doi.org/10.3390/jof8080841
Chicago/Turabian StylePan, Lili, Yuanyuan Yao, Hailin Zheng, Shuzhen Yan, and Shuanglin Chen. 2022. "Inhibitory Effects and Mechanism of Action of Elsinochrome A on Candida albicans and Its Biofilm" Journal of Fungi 8, no. 8: 841. https://doi.org/10.3390/jof8080841