CONICET | Buscador de Institutos y Recursos Humanos

Fungal bioactive compounds have a great potential to be used inthe medical field. In particular, different fungal proteins with immunomodulatory activity have been described. The aim of this work wasto obtain enriched protein extracts from fruiting bodies of Pleurotusostreatus (Po) and Agaricus bisporus (Ab) in order to evaluate theireffect on RAW 264.7 cells. To achieve this goal, fruiting bodies obtained from the local market were lyophilized and homogenized withTris-Glycine buffer pH 8.4. Homogenates were centrifuged and thesupernatants were treated with ice cold ethanol to precipitate soluble proteins. The precipitates were resuspended in phosphate buffer 50 mM, pH 7. Protein concentration was measured by Bradford method and a SDS-PAGE was performed to obtain the protein bandprofile. An MTT assay was carried out to assess cellular metabolicactivity. RAW 264.7 cells (100.000 cells/well) were seeded in 24-well culture plates and allowed to attach for 24 h. Culture mediumwas replaced with DMEM containing different protein concentrationof Ab or Po extracts, and cells were incubated at 37 °C for an additional 24 h. At the end of incubation, supernatants were collectedand stored at -20°C, and cells were washed with PBS and incubatedat 37 ºC for 30 min in the presence of MTT. Purple formazan crystals were solubilized with etanol and absorbance was measure. Fornitrites quantification, Griess assay was performed using the previously collected supernatants. The results showed that both extractsdiminished metabolic activity of RAW 264.7 in a dose-dependentmanner. In the case of Po extract, IC50 was less than 0,025 mg/ml,while with Ab extract the IC50 was 0,078 mg/ml. Regarding nitritesconcentration, none of the treatments showed differences with thecontrol. The results obtained showed that the fungal extracts usedhave an impact on RAW 264.7 cells, but more studies are needed inorder to completely understand their effect on these cells.