Morphological and biochemical changes during formocresol induced cell death in murine peritoneal macrophages: apoptotic and necrotic features
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The present study was conducted to investigate the role of Formocresol (FC)-induced apoptosis and necrotic cell death in murine peritoneal macrophages (pMø). Macrophages were cultured with 1:100 FC for 2 to 24 h. The viability (trypan blue assay), cell morphology (scanning electronic microscope), and apoptotic and necrotic indexes (light and fluorescent microscopy) were determined at different scheduled times. Simultaneously, the expressions of proteins related to stress, survival, and cell death were measured by western blotting. FC-exposed macrophages exhibited maximal apoptosis from 2 to 6 h, coincident with Bax overexpression (P < 0.001). Additionally, Bcl-xL showed maximal expression between 12 and 24 h suggesting its survival effect in pMø. The lowest pMø viability and the increment of the necrotic rate from 4 to 12 h were observed in accordance to Fas and Hsp60 overexpressions. In summary, all the experimental data suggest that two different pathways emerge in pMø exposed to FC, one leading Bax-dependent apoptosis (2–6 h) and the other one favoring necrosis (4–18 h), related to Fas-receptor and Hsp60 stress signal.
KeywordsMacrophages Formocresol Apoptosis Bax Necrosis Fas
This work was supported by grants from SEGCyT-UNNE (PI016/05) and CONICET (PIP 1302), Argentina. We thank PhD Gabriela Lucas for the clinical view of this study and Mrs Mirta Alba Alvarez for excellent technical assistance.
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