25º Congresso Brasileiro de Microbiologia

25º Congresso Brasileiro de Microbiologia

ResumoID:647-1

Área: Genética e Biologia Molecular ( Divisão N )

TRANSCRIPTIONAL ANALYSIS OF PARACOCCIDIOIDES BRASILIENSIS AFTER EXPOSITION TO OENOTHEIN B

Patricia F Zambuzzi (UFG); Renata S Prado (UFG); Raquel V Rezende (UFG); Clayton Luiz Borges (UFG); Pedro H Ferri (UFG); Suzana C Santos (UFG); Celia Maria de Almeida Soares (UFG); Maristela Pereira (UFG)

Resumo

Paracoccidioides brasiliensis is a causative agent of paracoccidioidomycosis (PCM), a human systemic mycosis geographically confined to Latin America. The long-time treatment, the high toxicity of the drugs, and the appearance of resistant or multi-resistant strains have imposed the need for a permanent search and development of new therapeutic approaches. Plants are biologically-active products, with great structural diversity, many of which are models for the synthesis of a large number of drugs. The active compound oenothein B, purified from leaves of Eugenia uniflora, a Brazilian Savannah plant, has been evaluated on the growth, viability and gene expression of P. brasiliensis. The compound interferes with yeast cell morphology and inhibits â-1,3-glucan synthase transcript. Aiming elucidate the mechanism of action of oenothein B on P. brasiliensis, was realized Representational Difference Analysis (RDA). P. brasiliensis yeast cells were grown on MMcM (Mc Veigh & Morton) medium in the presence and in the absence of oenothein B for 90 min, at 37^oC. After extraction of total RNA, the cDNA was obtained and used to RDA experiments, which originated 280 ESTs that were sequenced and originated 28 contigs and 24 singlets. By using BLATX program, the ESTs were classified in agreement with the functions. The analyses indicated the presence of transcripts with functions related to cell wall and membrane, elongation and transcription factors and hypothetic proteins. The synergism of oenothein B with drugs currently used in the treatment of PCM (amphotericin B, itraconazole, sulfamethoxazole and sulfametoxazol/trimetoprim) was also evaluated. The minimum inhibitory concentrations of all drugs were obtained and the test selective plate was carried. The growth of colonies of the fungus was measured after five days of incubation. The greatest inhibition occurred in the presence of sulfametoxazol/trimetoprim associated with oenothein B, followed by Amphotericin B associated with oenothein B, itraconazole associated with oenothein B and sulfamethoxazole associated with oenothein B, respectively. The elucidation of the action mechanism of oenothein B will facilitate the design and synthesis of related compounds with enhanced pharmacological profiles.

Palavras-chave: Paracoccidioides brasiliensis, Representational Diference analysis, oenothein B

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