612. Astrocyte-mediated inflammatory response as a promising target for TRP: a study in humanized Alzheimer’s disease mice model

Abstract

Abstract Background Chronic inflammation within the central nervous system is a defining feature of many neurological and neurodegenerative disorders, including Alzheimer’s disease (AD). Astrocytes, the most abundant glial cell population in the brain, play a pivotal role in maintaining neuronal homeostasis, synaptic support, and metabolic regulation. Under pathological conditions, astrocytes undergo transformation, characterized by extensive cytokine release and impaired homeostatic functions, which can amplify inflammatory signaling and contribute to neuronal dysfunction. Although non-steroidal anti-inflammatory drugs (NSAIDs) such as ibuprofen are commonly employed to suppress inflammatory pathways, their clinical utility in neurodegeneration is constrained by limited brain availability and dose-dependent adverse effects. This has driven increasing interest in the development of novel anti-inflammatory compounds capable of effectively modulating glial reactivity with improved safety and sustained central activity. Aims & Objectives The aim of this study was to assess the anti-inflammatory and pro-resolving effects of ibuprofen (IBU) and a novel compound (TRP) in immune-stimulated primary astrocyte cultures derived from C57BL/6J wild-type (WT) mice as well as APPNL-F/NL-F knock-in mice (humanized model of AD). Method Primary astrocytes were isolated from the cerebral cortices of 1–2-day-old mice and exposed to IBU or TRP (10 μM) for 1 h, followed by inflammatory stimulation with lipopolysaccharide (LPS; 100 ng/mL) for 24 h. Astrocytic viability was evaluated using the LDH release assay. Nitric oxide production was quantified using the Griess method, while the release of selected pro-inflammatory (IL-1β, IL-6, CCL2) and anti-inflammatory (IL-10, IL-4) cytokines was determined by ELISA. Results Treatment with both IBU and TRP significantly improved astrocyte survival under inflammatory conditions, as indicated by reduced LDH release. LPS exposure markedly increased nitric oxide production and the secretion of pro-inflammatory mediators, whereas TRP treatment effectively suppressed NO, IL-1β, and CCL2 levels in stimulated astrocytes. These effects were observed in cultures derived from both WT and AD-model mice. Discussion & Conclusions The present findings demonstrate that TRP attenuates astrocyte-mediated inflammatory responses and confers protection against LPS-evoked cellular damage. Hence, TRP emerges as a promising candidate for further investigation as a modulator of astrocyte-driven neuroinflammation and/or Alzheimer’s disease. 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).