PROTEOMIC PROFILE CHANGES IN CANDIDA ALBICANS TREATED WITH BARK EXTRACT OF STRYPHNODENDRON ADSTRINGENS

Tatiana Maria Souza Moreira (UNESP); Henrique Luiz Rodrigues da Cunha (UNIARA); Daniella Barros Rossetto (UNESP); Danuza Rossi (UNESP); Cleslei Fernando Zanelli (UNESP); Sandro Roberto Valentini (UNESP); Rosemeire Cristina Linhari Rodrigues Pietro (UNESP)

The barks of Stryphnodendron adstringens (Mart.) Coville is widely used in Brazil to treat several infections and wounds. Among its therapy uses, there are scientific evidences favoring its activity against candidiasis. However, it is unknown the means of the antimicrobial activity. The aim of this study was to analyze by SDS-PAGE, proteomic profile changes of Candida albicans treated with bark extract of S. adstringens compared with an untreated yeast sample. Bark extract was obtained with 70% ethanol, evaporated under pressure and lyophilized. C. albicans (ATCC 64548) was cultured in Sabouraud-agar for 48 h at 37° C. An adjusted suspension of the yeast was inoculated in a volume of 20 mL of YPD in order to achieve 2.5x10³ CFU/mL. One sample was treated with 312 mg/mL (minimum inhibitory concentration extract value) of extract and the other did not receive any treatment. After incubation at 35° C, 150 rpm, overnight, the suspension were centrifuged at 5000 g for 10 min at 4° C and washed with cold PBS twice. The pellet was dissolved in Tris buffer with protease inhibitor and received vigorous mechanical agitation for 1 min, six times. The protein of this supernatant was quantified by Lowry method in order to apply 50 mg of protein of each sample in a SDS-PAGE gel of electrophoresis (0.75 mm thickness; 4% stacking gel; 12.5% running gel) that was carried out at 150 V for about 2 h. Gels were stained with Coomassie blue G-250 (0.1% v/v), destained with 1% acetic acid and dried. Digitalized gel was analyzed by TotalLab TL120 Program and revealed 19 distinct bands of C. albicans untreated with 2 majority ones and 25 bands of the sample treated with S. adstringens extract presenting 3 majority ones. It was noted the absence of the bands of 83 and 66 kDa in the sample treated and the presence of 120, 113, 102, 92, 58 and 44 kDa bands that were not present in the untreated sample. This results showed significant changes in the proteomic profile of C. albicans when treated with bark extract of S. adstringens that could be involved in its action mechanism.