Fatemeh Jahansooz; Sara Keyhaniasl; Shamim Elahi; Ashraf Heidaripour
Abstract
Grapes and raisins with and without seeds were extracted using ethyl acetate/H2O solvent. The antioxidant activity of each extract was confirmed by the 1,1-diphenyl-2-picrylhydrazyl ...
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Grapes and raisins with and without seeds were extracted using ethyl acetate/H2O solvent. The antioxidant activity of each extract was confirmed by the 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical scavenging method and free radical scavenging assay. The results confirmed the antioxidant property of the extract in such a way that the most antioxidant activity of the extract was on radical activity. Free radicals were observed in extracts of seedless raisins and seeded grapes. Their cytotoxicity was investigated on cancer stem cells (SHSY5Y) using the MTT (p-value = <0.0001) method. The results obtained from the MTT test were confirmed using the cell counting method and Trypan blue staining. Cells (SHSY5Y-1) were identified with IC50=66.87 µg/mL in seeded grapes extract and IC50=12.63 µg/mL in raisins without seeds extract. The FITC-V Annexin+ and PI+ cell populations were considered as secondary apoptosis, and FITC-V Annexib- and PI+ cell populations as necrosis. Data analysis was done using device software. Flow cytometry analysis showed that 62.2 % of cells underwent cell apoptosis after treatment with seedless raisin extract and 78% of cells after treatment with seeded grapes. Real time PCR method was performed for the molecular analysis of treated cells. The change was evaluated in the expression of apoptotic genes (BAX, BCL2, P53, and GAPDH). Expression of apoptotic genes for BAX and BCL2 is high under treatment with seeded grape extract. However, the expression of apoptotic genes for BCL2 and P53 is high under treatment with raisin without grape extract.
