Neuroprotection and activation of anti-inflammatory and plasticity mechanisms by PLC-β inhibition after cholinergic hyperstimulation in hippocampal slices and microglial cells

Epilepsy, a chronic neurological disorder characterized by recurring seizures, involves inflammatory and abnormal plasticity features. Current treatments, primarily combinations of anticonvulsant drugs, often carry adverse effects, and a substantial portion of patients remain resistant to available therapies. In this study, we used the muscarinic agonist pilocarpine (Pilo) to probe epileptogenesis mechanisms and explore the effects of edelfosine (Ef), a synthetic alkyl-lysophospholipid (ALP), and phospholipase C (PLC)-β inhibitor, as a potential anti-epileptic agent. By targeting PLC-β activation, a link between muscarinic stimulation and brain injury, we assessed edelfosine's neuroprotective effects in an ex-vivo Pilo model. Ef, at 1 and 10 μM, significantly preserved neuronal PS in hippocampal slices compared to controls. Additionally, Ef (1 μM) in vitro studies demonstrated a reduction in proinflammatory gene expression, tumor necrosis factor-α (<i>TNF-α</i>), interlukin-6 (<i>IL-6</i>), and nuclear factor kappa-B1 (<i>NFκB1</i>) in microglial cells after exposure to different proinflammatory insults. Furthermore, it decreased the expression of aberrant plasticity genes, brain-derived neurotrophic factor (<i>BDNF</i>), matrix metalloproteinase 9 (<i>MMP9</i>), and serum response factor (<i>SRF</i>) implicated in epilepsy. Notably, these effects were observed without compromising cell viability. Our findings propose PLC-β inhibition as a promising therapeutic strategy against seizures, with Ef emerging as a potential non-classical treatment option.