Inhibitory effects of carvacrol on the expression of secreted aspartyl proteinases 1-3 in fluconazole-resistant Candida albicans isolates
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Abstract
Background and Objectives: Secreted Aspartyl Proteinase (SAP) is one of the main virulence factors in the pathogenesis ofCandida. This enzyme is encoded by a family of at least ten genes. Among these genes, the role of SAP1-3 in mucosal infections is evident. This study aimed to investigate the expression of SAP1-3 genes of Candida albicans isolates after treatmentwith Echinophora platyloba extract, carvacrol and caspofungin drug.
Materials and Methods: Vaginal samples of 68 women with suspected vaginitis were obtained and cultured. Canida albicansspecies were identified using phenotypic and genotyping methods. Spectrophotometry was used to investigate thepresence of SAP protein in the vaginal samples, and SDS-PAGE was used to confirm its protein composition. Real-timePCR was performed to ascertain the effects of subinhibitory concentrations of Echinophora platyloba extract, carvacrol andcaspofungin on the expression of SAP1-3 genes before and after treatment.
Results: C. albicans was found as the abundant species (59.6%), and different amounts of SAP were present in all vaginalsamples, which were higher than Candida krusei strain. The protein composition of SAP in C. albicans samples was estimatedwith the approximate molecular weight of 45 kDa. mRNA levels of total SAP in FLU-resistant isolates (P=0.01) were morethan those of FLU-susceptible isolates (P=0.07). The findings indicated that carvacrol is effective in reduction of SAP1-3 expression with a particular effect against FLU-resistant isolates.
Conclusion: Carvacrol contains an essential oil (carvacrol); therefore, it can be considered as an alternative effective antifungal compound.
Wisplinghoff H, Bischoff T, Tallent SM, Seifert H, Wenzel R, Edmond MB. Nosocomial bloodstream infections in US hospitals: analysis of 24,179 cases from a prospective nationwide surveillance study. Clin Infect Dis 2004;39:309-17.
Arendrup M.C, Rodriguez-Tudela J.L, Park S, Garcia-Effron G, Delmas G, Cuenca-Estrella M, et al. Echinocandin susceptibility testing of Candida spp. Using euast edef7.1 and CLSI M27-A3 standard procedures: Antimicrob Agents Chemother 2011b; 55:1580–1587.
Consolaro L, Albertoni A, Yoshida C, Mazucheli J. Correlation of Candida species and symptoms among patients with vulvovaginal candidiasis in Maringa . Iberoam Micol.2004; 21:202-205.
Jaeger M, Plantinga TS, Joosten LA, Kullberg BJ, Netea MG. Genetic basis for recurrent vulvo-vaginal candidiasis. J Curr Infect Dis Rep 2013;15: 136-42.
Naglik JR, Challacombe SJ, Hube B. Candida albicans secreted aspartyl proteinases in virulence and pathogenesis. J Microbiol Mol Biol Rev 2003;67:400-28.
He XY, Meurman JH, Kari K, Rautemaa R, Samaranayake LP. In vitro adhesion of Candida species to denture base materials. J Mycoses2006; 49:80-4.
Negri B, Silva N, Lopes L, Henriques M, Azeredo O, Oliveira K. An in vitro evaluation of Candida tropicalis infectivity using human cell monolayers. Journal of Medical Microbiology 2011; 60: 1270–1275.
Staib P, Kretschmar M, Nichterlein T, Hof H, Morschhauser J. Transcriptional Regulators Cph1p and Efg1p Mediate Activation of the Candida albicans Virulence Gene SAP5 during Infection . J Infect Immun 2002; 70: 921–927.
Taylor B, Staib P, Binder A, Biesemeier A, Sehnal M, llinghoff M et al. Profile of Candida albicans secreted aspartic proteinase elicited during vaginal infection. J Infection and Immunity 2005;1828–1835.
Schaller M, Schafer W, Korting HC, Hube B. Differential expression of secreted aspartyl proteinases in a model of human oral candidosis and in patient samples from the oral cavity. J Mol Microbiol 1998; 29: 605-15.
Hube B, Naglik J. Candida albicans proteinases: resolving the mystery of a gene family.J Microbiology 2001;147:1997-2005.
Vanden Bossche, H, Marichal P. Molecular mechanisms of drug resistance in fungi. Trends Microbiol 1994; 2:393–400.
Sanglard D, Ischer F, Marchetti O, Entenza J, Bille J. Calcineurin A of Candida albicans: involvement in antifungal tolerance, cell morphogenesis and virulence.J Mol Microbiol 2003;48:959-76.
McCormack PL, Perry CM. Caspofungin: a review of its use in the treatment of fungal infections. J Drugs 2005; 65:2049-68.
Pinto E, Vale-Silva L, Cavaleiro C, Salgueiro L. Antifungal activity of the clove essential oil from Syzygium aromaticum on Candida, Aspergillus and dermatophyte species. J Med Microbiol 2009;58:1454-62.
Dorman HJ, Deans SG. Antimicrobial agents from plants: antibacterial activity of plant volatile oils. J Appl Microbiol 2000; 88: 308-16.
Weber FJ, de Bont JA. Adaptation mechanisms of microorganisms to the toxic effects of organic solvents on membranes. J Biochim Biophys Acta.1996; 29:225-45.
Aslani P, Yadegari M, Rajabibazl M. Investigation the effect of Echinophora platyloba and Satureja bachtiarica on MDR1 and ERG11 gene expression in fluconazole resistance clinical isolates Candida albicans using Real- time PCR. European Journal of Experimental Biology 2014;41:375-379.
Facp J, Alexander B, Andes D, Arthington-Skaggs B, Brwn S et al. Reference Method for Broth Dilution Antifungal Susc eptibility Testing of Yeasts; Approved St andard—Third Edition 2008;14:273-289.
Rehani S, Rao N, Rao A, Carnelio S, Ramakrishnaiah S, Prakash P.K. Spectrophotometric analysis of the expression of secreted aspartyl proteinases from Candida in leukoplakia and oral squamous cell carcinoma. Journal of Oral Science 2011;53: 421-425.
Laemmli UK .Cleavage of structural proteins during the assembly of head of bacteriophage. Bio protocol 1970; 227: 680–685.
Dostal L, Hamal P, Pavlickova L, Soucek M , Ruml T, Pichova I, et al. Simple Method for Screening Candida Species Isolates for the Presence of Secreted Proteinases. J Clin Microbiol 2003; 41:712–716.
Cutfield S, Marshall C, Moody P, Sullivian F, Cutfield J. Crystallization of inhibited aspartic proteinase from Candida albicans.. J Mol Biol 1993; 234: 1266-1269.
Zaugg C, Borg-von Zepelin M, Reichard U, Sanglard D, Monod M. Secreted aspartic proteinase family of Candida tropicalis. J Infection and Immunity 2001; 69:405-412.
Naglik JR., Moyes D, Makwana J, Kanzaria P, Tsichlaki E, Weindl G, et al .(2008). Quantitative expression of the Candida albicans secreted aspartyl proteinase gene family in human oral and vaginal candidiasis. J Microbiology 2008;154: 3266-80.
Monroy-Ramirez HC, Basurto-Islas G, Mena R, Cisneros B, Binder LI, Avila J, et al. J Alzheimers Dis 2013;36: 503-20.
WU T, Samaranayake L.P, Leung W.K, Sullivan P.A. Inhibition of growth and secreted aspartyl proteinase production in Candida albicans by lysozyme. J Med Microbiol1999; 48 :721-730.
Schaller M, Korting H.C, Borelli C, Hamm G, Hube B. Candida albicans secreted aspartic proteinases modify the epithelial cytokine response in an in vitro model of vaginal candidiasis. J Infect Immune 2005;73: 2758–2765.
Cadicamo CD, Mortier J, Wolber G, Hell M, Heinrich IE, Michel D, et al. Design, synthesis, inhibition studies, and molecular modeling of pepstatin analogues addressing different secreted aspartic proteinases of Candida albicans. Biochem Pharmacol 2013;85: 881-887.
Lieke A, Burt S, Veenendaal A, Bleumink N, Putten J. The Natural Antimicrobial Carvacrol Inhibits Campylobacter jejuni Motility and Infection of Epithelial Cells.J PMCID 2012;7:45343.
Reddy K.H, Govender P, Reddy O.V, Sarma P.V. Effect of hexane extract of Dillenia indica seed on the activity of secreted aspartyl proteinase of Candida albicans and its kinetic studies. Journal of Medicinal Plants Research 2012; 6:595-560.
Cho O, Shiokama T, Ando Y, Aoki N, Uehara C, Maeda E, Matsumoto S, Kurakado S and Sugita T. Screening of Compounds from an FDA-Approved Drug Library for the ability to Inhibit aspartic protease secretion from the Pathogenic Yeast Candida albicans. Pharmaceut Reg Affairs 2014; 3: 2167-7689.
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| Issue | Vol 8 No 6 (2016) | |
| Section | Articles | |
| Keywords | ||
| Carvacrol SAP1-3 Candida albicans Fluconazole-resistant Echinophora platyloba | ||
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