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| Author(s): |
Irlan de Almeida Freires
Total Authors: 1
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| Document type: | Master's Dissertation |
| Press: | Piracicaba, SP. |
| Institution: | Universidade Estadual de Campinas (UNICAMP). Faculdade de Odontologia de Piracicaba |
| Defense date: | 2014-02-26 |
| Examining board members: |
Pedro Luiz Rosalen;
Severino Matias de Alencar;
Bruno Bueno Silva
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| Advisor: | Pedro Luiz Rosalen; Marta Cristina Teixeira Duarte |
| Abstract | |
Introduction: Oral candidiasis is a common opportunistic fungal infection of the oral cavity with increasingly significant worldwide prevalence and incidence rates. Accordingly, novel specific-targeted strategies to manage this ailment have been proposed, among which is the use of essential oils (EO) with antifungal properties. Coriandrum sativum L. (coriander) EO has proven antifungal activity against Candida species and thus deserves further investigation. Objective: This study aimed to evaluate the antifungal activity and mode of action of the EO from Coriandrum sativum L. leaves on Candida spp., and to determine the molecular targets affected in the whole-genome expression in human cells. Material and Methods: C. sativum was obtained from the Germoplasm Bank of the Research Center for Chemistry, Biology and Agriculture (University of Campinas, SP, Brazil) whose EO and active fraction had their phytochemical profile determined by Gas Chromatography coupled to Mass Spectrometry. Then, we carried out the following tests with five reference strains of Candida spp. (CBS): Minimum Inhibitory and Fungicidal Concentration (MIC/MFC); antifungal mode of action (sorbitol and ergosterol assays); Scanning Electron Microscopy (SEM) analysis of Candida biofilm and tests of inhibition of biofilm adherence. We used nystatin, amphotericin B or caspofungin as positive controls, in addition to negative controls. Also, we tested the effect of C. sativum EO on the proteolytic activity of C. albicans. Next, a pharmacogenomic assay identified which molecular targets in the human genome were affected by C. sativum EO and its active fraction. Tests were performed in triplicate of independent experiments and data were statistically treated (ANOVA, Tukey¿s post-test, ?=0.05). Pharmacogenomic data were processed on GeneGo MetaCore® and DAVID Bioinformatics Resources. Results: The EO phytochemical profile indicated monoterpenes (37.9 %) and sesquiterpenes (62.1 %) as major compounds. The MIC/MFC values for the EO ranged from 15.6 to 62.5 µg/ml. With regard to the mode of action, C. sativum EO may bind to membrane ergosterol, increasing ionic permeability and causing membrane damage to cell death, but it does not act on cell wall biosynthesis-related pathways. This mode of action confirms the changes in the integrity of the biofilm morphology as verified in the analyses by SEM. The EO showed anti-adherent activity at low concentrations (31.2 ¿ 62.5 µg/ml) against all strains tested, as well as activity against proteases produced by C. albicans, with statistical significance at MIC (P < 0.05). Finally, the EO and its active fraction had low cytotoxicity on human cells with IC30 of 359.8 and 366.7 µg/ml, respectively, affecting the pathways of chemokines and MAP-kinase (apoptosis, proliferation), as well as adhesion proteins. Conclusions: The EO from C. sativum leaves has strong antifungal and anti-adherent activity against Candida spp., as well as anti-proteolytic activity against C. albicans, and acts by increasing cell membrane ionic permeability, probably due to the synergistic effect of mono- and sesquiterpenes. Pharmacogenomic analyses revealed low cytotoxicity with specific targets affected in the human genome, which encourages further pre-clinical toxicological screening and clinical research in this field (AU) | |
| FAPESP's process: | 11/15984-0 - Anti-Candida, antiproliferative and cytotoxic activities evaluation of essential oil and fractions from Coriandrum sativum L. |
| Grantee: | Irlan de Almeida Freires |
| Support Opportunities: | Scholarships in Brazil - Master |