A Survey of the Effect of Camphor on INT1 and EFG1 Gene Expressions of Candida albicans at Three Treatment Times (24, 48, and 72 hours) via Real-time PCR

Authors
R. Ghaffaripour 1
M. Rajabi Bazl 2
M. H. Yadegari 1
1 Department of Medical Mycology, Faculty of Medicine, Tarbiat Modares University, Tehran, Iran
2 Department of Clinical Biochemistry, Faculty of Medicine, Shahid Beheshti University of Medical Sciences, Tehran, Iran
Abstract
Objective: Candida albicans (C. albicans) is an opportunistic yeast that can lead to pathogenesis in immunocompromised individuals and under suitable conditions. Medicinal plants ingredients such as camphor can reduce the expressions of genes involved in virulence of the fungi through their antifungal properties. The products of INT1 and EFG1 are implicated in inducing filamentous growth and adhesion of C. albicans to the host tissues. Both of these characteristics are very important in its virulence. The present study focuses on the evaluation of the effects of camphor on INT1 and EFG1 expressions at three time points of treatment (24, 48, and 72 hours) via real-time PCR.
Methods: We prepared serial dilutions of camphor (1, 2, 4, 8, 16, 32, 64, 128, 256, and 512 mg/ml) and co-cultured them with 1.5×103 cells/ml of a C. albicans ATCC 10231 suspension for 48 hours at 350C. Next, we determined the MIC50/90 and MFC. C. albicans cells were treated with the MIC50 concentration of camphor for 24, 48, and 72 hours. RNA from C. albicans was extractedbefore and after treatment, back translated into cDNA, and analyzed with real-time PCR.
Results:MIC50, MIC90 and MFC of camphor were determined at 16 mg/ml, 32 mg/ml and 64 mg/ml, respectively. Evaluation of gene expression changes in yeast showed that camphor reduced the INT1 gene expression about 87% at 24 hours, 97% at 48 hours, and 86% at 72 hours after treatment compared to the untreated sample. EFG1 expression reduced about 58% at 24 hours, 93% at 48 hours, and 49% at 72 hours after treatment with camphor.
Conclusion: In recent years, advancements have been seen in herbalism due to the increased drug resistance and adverse effects of chemical drugs. These plants may efficiently act as antifungal agents. The results of this study have shown that the use of camphor can significantly reduce the expression of virulence genes INT1 and EFG1 in C. albicans.

Keywords

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Pathobiology Reserach
Volume 19, Issue 3
November 2016
Pages 59-72