Berberine Antifungal Activity in Fluconazole-Resistant Pathogenic Yeasts: Action Mechanism Evaluated by Flow Cytometry and Biofilm Growth Inhibition in Candida spp

The incidence of fungal infections and, in particular, the incidence of fungal antibiotic resistance, which is associated with biofilm formation, have significantly increased, contributing to morbidity and mortality. Thus, new therapeutic strategies need to be developed. In this context, natural pro...

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Published in:	Antimicrobial agents and chemotherapy Vol. 60; no. 6; pp. 3551 - 3557
Main Authors:	da Silva, Anderson Ramos, de Andrade Neto, João Batista, da Silva, Cecília Rocha, Campos, Rosana de Sousa, Costa Silva, Rose Anny, Freitas, Daniel Domingues, do Nascimento, Francisca Bruna Stefany Aires, de Andrade, Larissa Nara Dantas, Sampaio, Letícia Serpa, Grangeiro, Thalles Barbosa, Magalhães, Hemerson Iury Ferreira, Cavalcanti, Bruno Coêlho, de Moraes, Manoel Odorico, Nobre Júnior, Hélio Vitoriano
Format:	Journal Article
Language:	English
Published:	United States American Society for Microbiology 01-06-2016
Subjects:	Animals Antifungal Agents Antifungal Agents - pharmacology Berberine Berberine - adverse effects Berberine - pharmacology Berberis Berberis aquifolium Berberis vulgaris Biofilms - growth & development Candida Candida - classification Candida - drug effects Candida - genetics Candidiasis Candidiasis - drug therapy Candidiasis - microbiology Cell Line Cell Proliferation Cryptococcosis Cryptococcosis - drug therapy Cryptococcosis - microbiology Cryptococcus neoformans Cryptococcus neoformans - classification Cryptococcus neoformans - drug effects Cryptococcus neoformans - genetics DNA, Fungal - genetics Drug Resistance, Fungal Fluconazole Fluconazole - adverse effects Fluconazole - pharmacology Humans Hydrastis canadensis L Cells (Cell Line) Mechanisms of Action: Physiological Effects Mice Microbial Sensitivity Tests Mitochondrial Membranes - drug effects Molecular Typing Mycological Typing Techniques Phellodendron
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Summary:	The incidence of fungal infections and, in particular, the incidence of fungal antibiotic resistance, which is associated with biofilm formation, have significantly increased, contributing to morbidity and mortality. Thus, new therapeutic strategies need to be developed. In this context, natural products have emerged as a major source of possible antifungal agents. Berberine is a protoberberine-type isoquinoline alkaloid isolated from the roots, rhizomes, and stem bark of natural herbs, such as Berberis aquifolium, Berberis vulgaris, Berberis aristata, and Hydrastis canadensis, and of Phellodendron amurense Berberine has been proven to have broad antibacterial and antifungal activity. In the present study, the potential antifungal effect of berberine against fluconazole-resistant Candida and Cryptococcus neoformans strains, as well as against the biofilm form of Candida spp., was assessed. The antifungal effect of berberine was determined by a broth microdilution method (the M27-A3 method of the Clinical and Laboratory Standards Institute) and flow cytometry techniques, in which the probable mechanism of action of the compound was also assessed. For biofilm assessment, a colorimetric 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2H-tetrazolium bromide (MTT) assay was used to determine the susceptibility of sessile cells. The isolates used in the study belonged to the Laboratory of Bioprospection and Experiments in Yeast (LABEL) of the Federal University of Ceará. After 24 and 72 h, fluconazole-resistant Candida and Cryptococcus neoformans strains showed berberine MICs equal to 8 μg/ml and 16 μg/ml, respectively. Cytometric analysis showed that treatment with berberine caused alterations to the integrity of the plasma and mitochondrial membranes and DNA damage, which led to cell death, probably by apoptosis. Assessment of biofilm-forming isolates after treatment showed statistically significant reductions in biofilm cell activity (P < 0.001).
Bibliography:	ObjectType-Article-1 SourceType-Scholarly Journals-1 ObjectType-Feature-2 content type line 23 Citation da Silva AR, de Andrade Neto JB, da Silva CR, Campos RDS, Costa Silva RA, Freitas DD, do Nascimento FBSA, de Andrade LND, Sampaio LS, Grangeiro TB, Magalhães HIF, Cavalcanti BC, de Moraes MO, Nobre Júnior HV. 2016. Berberine antifungal activity in fluconazole-resistant pathogenic yeasts: action mechanism evaluated by flow cytometry and biofilm growth inhibition in Candida spp. Antimicrob Agents Chemother 60:3551–3557. doi:10.1128/AAC.01846-15.
ISSN:	0066-4804 1098-6596
DOI:	10.1128/AAC.01846-15