Details

Autor(en) / Beteiligte

• Couturier, J
• Paccalin, M
• Morel, M
• Terro, F
• Milin, S
• Pontcharraud, R
• Fauconneau, B
• Page, G

Titel

Prevention of the [beta]-amyloid peptide-induced inflammatory process by inhibition of double-stranded RNA-dependent protein kinase in primary murine mixed co-cultures

Ist Teil von

• Journal of neuroinflammation, 2011-06, Vol.8, p.72

Ort / Verlag

BioMed Central Ltd

Erscheinungsjahr

2011

Quelle

EZB Electronic Journals Library

Beschreibungen/Notizen

• Background Inflammation may be involved in the pathogenesis of Alzheimer's disease (AD). There has been little success with anti-inflammatory drugs in AD, while the promise of anti-inflammatory treatment is more evident in experimental models. A new anti-inflammatory strategy requires a better understanding of molecular mechanisms. Among the plethora of signaling pathways activated by [beta]-amyloid (A[beta]) peptides, the nuclear factor-kappa B (NF-[kappa]B) pathway could be an interesting target. In virus-infected cells, double-stranded RNA-dependent protein kinase (PKR) controls the NF-[kappa]B signaling pathway. It is well-known that PKR is activated in AD. This led us to study the effect of a specific inhibitor of PKR on the A[beta]42-induced inflammatory response in primary mixed murine co-cultures, allowing interactions between neurons, astrocytes and microglia. Methods Primary mixed murine co-cultures were prepared in three steps: a primary culture of astrocytes and microglia for 14 days, then a primary culture of neurons and astrocytes which were cultured with microglia purified from the first culture. Before exposure to A[beta] neurotoxicity (72 h), co-cultures were treated with compound C16, a specific inhibitor of PKR. Levels of tumor necrosis factor-[alpha] (TNF[alpha]), interleukin (IL)-1[beta], and IL-6 were assessed by ELISA. Levels of P.sub.T451 -PKR and activation of I[kappa]B, NF-[kappa]B and caspase-3 were assessed by western blotting. Apoptosis was also followed using annexin V-FITC immunostaining kit. Subcellular distribution of P.sub.T451 -PKR was assessed by confocal immunofluorescence and morphological structure of cells by scanning electron microscopy. Data were analysed using one-way ANOVA followed by a Newman-Keuls' post hoc test Results In these co-cultures, PKR inhibition prevented A[beta]42-induced activation of I[kappa]B and NF-[kappa]B, strongly decreased production and release of tumor necrosis factor (TNF[alpha]) and interleukin (IL)-1[beta], and limited apoptosis. Conclusion In spite of the complexity of the innate immune response, PKR inhibition could be an interesting anti-inflammatory strategy in AD.