Trans-chalcone and quercetin down-regulate fatty acid synthase gene expression and reduce ergosterol content in the human pathogenic dermatophyte Trichophyton rubrum

BMC Complementary and Alternative Medicine - Tập 13 Số 1 - 2013
Tamires Aparecida Bitencourt1, Tatiana Takahasi Komoto1, Bruna Gabriele Massaroto1, Carlos Eduardo Saraiva Miranda1, René Oliveira Beleboni1, Mozart Marins1, Ana Lúcia Fachin1
1Unidade de Biotecnologia, Universidade de Ribeirão Preto, Av: Costábile Romano 2201, 14096-900, Ribeirão Preto, SP, Brazil

Tóm tắt

Abstract Background Fatty acid synthase (FAS) is a promising antifungal target due to its marked structural differences between fungal and mammalian cells. The aim of this study was to evaluate the antifungal activity of flavonoids described in the scientific literature as FAS inhibitors (quercetin, trans-chalcone, ellagic acid, luteolin, galangin, and genistein) against the dermatophyte Trichophyton rubrum and their effects on fatty acid and ergosterol synthesis. Methods The antifungal activity of the natural products was tested by the microdilution assay for determination of the minimum inhibitory concentration (MIC). The effect of the compounds on the cell membrane was evaluated using a protoplast regeneration assay. Ergosterol content was quantified by spectrophotometry. Inhibition of FAS by flavonoids was evaluated by an enzymatic assay to determine IC50 values. Quantitative RT-PCR was used to measure transcription levels of the FAS1 and ERG6 genes involved in fatty acid and ergosterol biosynthesis, respectively, during exposure of T. rubrum to the flavonoids tested. Results The flavonoids quercetin and trans-chalcone were effective against T. rubrum, with MICs of 125 and 7.5 μg/mL for the wild-type strain (MYA3108) and of 63 and 1.9 μg/mL for the ABC transporter mutant strain (ΔTruMDR2), respectively. The MICs of the fluconazole and cerulenin controls were 63 and 125 μg/mL for the wild-type strain and 30 and 15 μg/mL for the mutant strain, respectively. Quercetin and trans-chalcone also reduced ergosterol content in the two strains, indicating that interference with fatty acid and ergosterol synthesis caused cell membrane disruption. The MIC of quercetin reduced the number of regenerated protoplasts by 30.26% (wild-type strain) and by 91.66% (mutant strain). Half the MIC (0.5 MIC) of quercetin did not reduce the number of regenerated wild-type fungal colonies, but caused a 36.19% reduction in the number of mutant strain protoplasts. In contrast, the MIC and 0.5 MIC of trans-chalcone and cerulenin drastically reduced protoplast regeneration in the two strains. The FAS1 gene was repressed in the presence of MICs of quercetin, trans-chalcone, fluconazole and cerulenin. The ERG6 gene was induced in the presence of MICs of fluconazole and cerulenin and was repressed in the presence of MICs of trans-chalcone and quercetin. Trans-chalcone and quercetin inhibited the enzymatic activity of FAS, with IC50 values of 68.23 and 17.1 μg/mL, respectively. Conclusion Trans-chalcone and quercetin showed antifungal activity against T. rubrum, reducing ergosterol levels and modulating the expression of FAS1 and ERG6.

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Tài liệu tham khảo

Marconi VC, Kradin R, Marty FM, Hospenthal DR, Kotton CN: Disseminated dermatophytosis in a patient with hereditary hemochromatosis and hepatic cirrhosis: case report and review of the literature. Med Mycol. 2010, 48 (3): 518-527. 10.3109/13693780903213512.

Ameen M: Epidemiology of superficial fungal infections. Clinics in dermatology. 2010, 28 (2): 197-201.

Chen BK, Friedlander SF: Tinea capitis update: a continuing conflict with an old adversary. Curr Opin Pediatr. 2001, 13 (4): 331-335. 10.1097/00008480-200108000-00008.

Baddley JW, Moser SA: Emerging fungal resistance. Clin Lab Med. 2004, 24 (3): 721-735. 10.1016/j.cll.2004.05.003.

Projan SJ, Youngman PJ: Antimicrobials: new solutions badly needed. Curr Opin Microbiol. 2002, 5 (5): 463-465. 10.1016/S1369-5274(02)00364-8.

Roemer T, Xu D, Singh SB, Parish CA, Harris G, Wang H, Davies JE, Bills GF: Confronting the challenges of natural product-based antifungal discovery. Chem Biol. 2011, 18 (2): 148-164. 10.1016/j.chembiol.2011.01.009.

Monk BC, Cannon RD: Genomic pathways to antifungal discovery. Curr Drug Targets: Infect Disord. 2002, 2 (4): 309-329. 10.2174/1568005023342344.

Sturtevant J: Translation elongation-3-like factors: are they rational antifungal targets?. Expert Opin Ther Tar. 2002, 6 (5): 545-553. 10.1517/14728222.6.5.545.

Leibundgut M, Maier T, Jenni S, Ban N: The multienzyme architecture of eukaryotic fatty acid synthases. Curr Opin Struct Biol. 2008, 18 (6): 714-725. 10.1016/j.sbi.2008.09.008.

Maier T, Leibundgut M, Boehringer D, Ban N: Structure and function of eukaryotic fatty acid synthases. Q Rev Biophys. 2010, 43 (3): 373-422. 10.1017/S0033583510000156.

Nomura S, Horiuchi T, Omura S, Hata T: he action mechanism of cerulenin. I. Effect of cerulenin on sterol and fatty acid biosynthesis in yeast. J Biochem. 1972, 71 (5): 783-796.

Li XC, Joshi AS, ElSohly HN, Khan SI, Jacob MR, Zhang Z, Khan IA, Ferreira D, Walker LA, Broedel SE: Fatty acid synthase inhibitors from plants: isolation, structure elucidation, and SAR studies. J Nat Prod. 2002, 65 (12): 1909-1914. 10.1021/np020289t.

Zhang W, Yu L, Leng W, Wang X, Wang L, Deng X, Yang J, Liu T, Peng J, Wang J: cDNA microarray analysis of the expression profiles of Trichophyton rubrum in response to novel synthetic fatty acid synthase inhibitor PHS11A. Fungal genetics and biology: FG & B. 2007, 44 (12): 1252-1261. 10.1016/j.fgb.2007.03.002.

Yu L, Zhang W, Liu T, Wang X, Peng J, Li S, Jin Q: Global gene expression of Trichophyton rubrum in response to PH11B, a novel fatty acid synthase inhibitor. J Applied Microbiol. 2007, 103 (6): 2346-2352. 10.1111/j.1365-2672.2007.03521.x.

Fachin AL, Ferreira-Nozawa MS, Maccheroni W, Martinez-Rossi NM: Role of the ABC transporter TruMDR2 in terbinafine, 4-nitroquinoline N-oxide and ethidium bromide susceptibility in Trichophyton rubrum. J Med Microbiol. 2006, 55 (Pt 8): 1093-1099.

Arthington-Skaggs BA, Jradi H, Desai T, Morrison CJ: Quantitation of ergosterol content: novel method for determination of fluconazole susceptibility of Candida albicans. J Clin Microbiol. 1999, 37 (10): 3332-3337.

Cove DJ: The induction and repression of nitrate reductase in the fungus Aspergillus nidulans. Biochimica et biophysica acta. 1966, 113 (1): 51-56. 10.1016/S0926-6593(66)80120-0.

Nowakowska Z: A review of anti-infective and anti-inflammatory chalcones. Eur J Med Chem. 2007, 42 (2): 125-137. 10.1016/j.ejmech.2006.09.019.

Boeck P, Leal PC, Yunes RA, Filho VC, Lopez S, Sortino M, Escalante A, Furlan RL, Zacchino S: Antifungal activity and studies on mode of action of novel xanthoxyline-derived chalcones. Arch Pharm. 2005, 338 (2–3): 87-95.

Brown AK, Papaemmanouil A, Bhowruth V, Bhatt A, Dover LG, Besra GS: Flavonoid inhibitors as novel antimycobacterial agents targeting Rv0636, a putative dehydratase enzyme involved in Mycobacterium tuberculosis fatty acid synthase II. Microbiology. 2007, 153 (Pt 10): 3314-3322.

Jung HJ, Park Y, Sung WS, Suh BK, Lee J, Hahm K-S, Lee DG: Fungicidal effect of pleurocidin by membrane-active mechanism and design of enantiomeric analogue for proteolytic resistance. Biochimica et Biophysica Acta (BBA) - Biomembranes. 2007, 1768 (6): 1400-1405. 10.1016/j.bbamem.2007.02.024.

Lee DG, Park Y, Kim HN, Kim HK, Kim PI, Choi BH, Hahm K-S: Antifungal Mechanism of an Antimicrobial Peptide, HP (2–20), Derived from N-Terminus of Helicobacter pylori Ribosomal Protein L1 against Candida albicans. Biochem Biophys Res Communications. 2002, 291 (4): 1006-1013. 10.1006/bbrc.2002.6548.

Young LY, Hull CM, Heitman J: Disruption of ergosterol biosynthesis confers resistance to amphotericin B in Candida lusitaniae. Antimicrobial agents and chemotherapy. 2003, 47 (9): 2717-2724. 10.1128/AAC.47.9.2717-2724.2003.

Zhang W, Yu L, Yang J, Wang L, Peng J, Jin Q: Transcriptional profiles of response to terbinafine in Trichophyton rubrum. Appl Microbiol Biotechnol. 2009, 82 (6): 1123-1130. 10.1007/s00253-009-1908-9.

Barker KS, Crisp S, Wiederhold N, Lewis RE, Bareither B, Eckstein J, Barbuch R, Bard M, Rogers PD: Genome-wide expression profiling reveals genes associated with amphotericin B and fluconazole resistance in experimentally induced antifungal resistant isolates of Candida albicans. J Antimicrob Chemother. 2004, 54 (2): 376-385. 10.1093/jac/dkh336.

Leber R, Zinser E, Hrastnik C, Paltauf F, Daum G: Export of steryl esters from lipid particles and release of free sterols in the yeast, Saccharomyces cerevisiae. Biochimica et Biophysica Acta (BBA) - Biomembranes. 1995, 1234 (1): 119-126. 10.1016/0005-2736(94)00270-Y.

Pinto WJ, Lozano R, Nes WR: Inhibition of sterol biosynthesis by ergosterol and cholesterol in Saccharomyces cerevisiae. Biochimica et Biophysica Acta (BBA) - Lipids and Lipid Metabolism. 1985, 836 (1): 89-95. 10.1016/0005-2760(85)90224-3.

Servouse M, Karst F: Regulation of early enzymes of ergosterol biosynthesis in Saccharomyces cerevisiae. Biochem J. 1986, 240 (2): 541-547.

Casey WM, Keesler GA, Parks LW: Regulation of partitioned sterol biosynthesis in Saccharomyces cerevisiae. J Bacteriol. 1992, 174 (22): 7283-7288.

Smith SJ, Crowley JH, Parks LW: Transcriptional regulation by ergosterol in the yeast Saccharomyces cerevisiae. Mol Cell Biol. 1996, 16 (10): 5427-5432.

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BMC Complementary and Alternative Medicine

Tập 13 Số 1

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