Induction of autophagy in BV2 mciroglial cells modulates pro-inflammatory mediators levels and rescues both LPS and alpha-synuclein-induced neuronal cell death.

BUSSI C; PERALTA-RAMOS JAVIER M; GAVIGLIO E; ARROYO DS; GALLEA JI; CELEJ MS; IRIBARREN P

Congreso; IV LASID Meeting; 2015

Autophagy is a fundamental cellular homeostatic mechanism, whereby cells autodigest parts of their cytoplasm for removal or turnover. Paradoxically, although autophagy is primarily a protective process for the cell, it can also play a role in cell death.Microglial cells (MC) are resident macrophages in the central nervous system (CNS) and have multiple functions, such as phagocytosis, production of growth factors and cytokines, and antigen presentation. The aim of this study was to evaluate the effects of autophagy on the production of pro-inflammatory mediators by BV2 MC, and on neuronal viability in a co-culture model.Autophagy was induced in MC before or after TLR or alpha-synuclein (α-syn) stimulation by rapamycin or trehalose and blocked by using 3-Methyladenine. Supernatants were isolated and analyzed by ELISA and Griess assay to determine cytokines and nitric oxide (NO) levels, respectively. Autophagic activity was followed by confocal microscopy and WB. Cell death was evaluated using AnnexinV/propidium iodide staining and subsequent flow cytometric analysis.Autophagy induction in BV2 cells before LPS or α-syn stimulation downregulated IL1b, IL-6, TNFa and nitric oxide production (p<0.01). Furthermore, we observed in BV2/N2A co-cultures stimulated with LPS or α-syn fibers that induction of autophagy in microglial cells rescued LPS and α-syn-induced neuronal cell death (p<0.05).These results suggest that modulation of microglial cells by autophagy could be an important strategy in the context of neurodegenerative diseases.