613. The analysis of the impact of non-steroidal drug and the TRP compound on the immune profile of microglia: study in an Alzheimer’s disease model
Abstract Background Neuroinflammation is a hallmark of central nervous system (CNS) disorders and plays a critical role in the progression of neurodegenerative diseases, including Alzheimer’s disease. Microglia, the resident immune cells of the CNS, act as key regulators of inflammatory responses and are rapidly activated in response to pathological stimuli such as amyloid-β accumulation, neuronal injury, and synaptic dysfunction. While acute microglial activation is essential for tissue homeostasis and debris clearance, sustained or dysregulated activation contributes to chronic inflammation and neurodegeneration. Non-steroidal anti-inflammatory drugs (NSAIDs), including ibuprofen, are widely used to attenuate neuroinflammatory processes. Although ibuprofen exhibits anti-inflammatory and neuroprotective properties, achieving therapeutic concentrations in the brain requires prolonged dosing, which exacerbates various peripheral and central side effects. Therefore, there is a critical need to develop novel anti-inflammatory compounds with improved brain bioavailability and safer therapeutic profiles in order to effectively modulate neuroinflammation and in consequently, neurodegeneration. Aims & Objectives We propose as a goal of the present study a comparative evaluation of the anti-inflammatory and pro-resolution potential of ibuprofen (IBU) and a novel compound (TRP) in immune-activated primary microglial cell cultures derived from C57BL/6J (WT) and APPNL-F/NL-F knock-in mice (KI, animal model of AD). Method Primary Microglial cells were prepared from the cortices of 1-2-day-old mouse brains. The cells were treated for 1 h with IBU or TRP (10 μM) and then stimulated for 24 h with bacterial endotoxin (lipopolysaccharide, LPS, 100 ng/mL). We measured the cell death parameters (lactate dehydrogenase, LDH assay), the secretion of nitric oxide (NO) synthesis by Griess reaction, and pro- (IL-1β, IL-6, CCL2) and anti-inflammatory cytokines (IL-10, IL-4) profile release in control and AD-derived microglia cells using ELISA kits. Results Exposure to IBU and TRP significantly reduced LPS-induced microglial cell death, as evidenced by decreased LDH release. Moreover, LPS stimulation led to an upregulation of NO, and IL-1β, IL-6, and IL-10 release as well as CCL-2 production, more evident in microglia cells derived from KI mice. Most of the pro-inflammatory markers released were attenuated by TRP administration. Interestingly, TRP treatment preserved the LPS-induced elevation of IL-10, which may suggest not only an anti-inflammatory but also a pro-resolving profile of TRP action in primary microglia cultures obtained also from the AD model. Discussion & Conclusions Our findings suggest that TRP modulates microglial activation in a varied manner. Its simultaneous ability to influence the proinflammatory and anti-inflammatory profiles of microglial cells in an AD model positions this compound as a promising candidate for further investigation in the context of neuroimmune dysregulation and a tool for pharmacotherapy of resolution. The study was supported by a programme coordinated by the Medical Research Agency, co-financed by the European Union under the NextGeneration EU initiative, within the framework of the National Recovery Plan, Component D, Investment D3.1.1 (project no. 2024/ABM/03/KPO/KPOD.07.07-IW.07-0173/24-00).