The effects of ethanol extract propolis on extrinsic and intrinsic pathways of apoptosis in primary rat astrocyte cell culture

Presented study was to examine the effect of ethanol extract of propolis (EEP), dependently on their concentrations on the viability and morphology of exposition to treated EEP or its ingredients. Results about 100μg/ml ingredients of EEP were evaluated by Immumoblot. We obtained the strong decline of viability of primary astrocytes cell. The results of Immunoblot shown an increasing of GFAP expression in primary astrocyte cell culture approximately to 29% after EEP 10μg/ml, 22% after EEP 25μg/ml and 79% after EEP 100μg/ml application. Moreover, the modulation of expression markers of apoptotic ways, especially, caspase3, BAX, Bcl-2 may indicate activation of intrinsic pathway of apoptosis induced by EEP in primary astrocyte cell culture line. EEP contain high amount antioxidant known to be essential for maintaining the proper function and regulation of many biochemical and cellular reactions. However, high exposure to levels of EEP in occupational can lead to more degenerative suppression caspase-3,BAX. Related to apoptosis, according to caspase-3 content on 79%, BAX 66% (EEP+LPS). Observed results showed that primary astrocyte cell culture is valid experimental model for the study of EEP effects on switching off apoptosis pathways. Different doses of EEP have a non linear dose-depend effect on expressions of both extrinsic and intrinsic apoptotic pathways markers. After treatment with EEP primary astrocyte cell culture induce astroglial reactivity in 10–100 μg/ml range concentration. This astrocyte‟s reactivation associated with GFAP overexpression and hypertrophy of glial cells. The involvement of the intrinsic mitochondrial pathway of apoptosis was demonstrated with modulation by EEP a level of Bcl-2 expression. In addition, our data also shown for the first time that 25μg/ml EEP induce maximum astroglial reactivity compared with 10 – 100μg/ml EEP. Taking together, these findings contribute to a deeper elucidation of the molecular signaling mechanisms underlying EEP for regulation apoptotic pathways and cell reactivation.