<p>N-acylethanolamines (NAEs) derived from omega-3 polyunsaturated fatty acids (PUFAs), including N-docosahexaenoylethanolamine (DHEA, FA 22:6(n-3)-EA), N-eicosapentaenoylethanolamine (EPEA, FA 20:5(n-3)-EA), and N-stearidonoylethanolamine (SDEA, FA 18:4(n-3)-EA), exhibit anti-inflammatory and antioxidant properties, making them promising candidates for the modulation of neuroinflammation. This study investigated their effects on LPS-stimulated SIM-A9 microglial cells, focusing on cytotoxicity, inflammatory markers, oxidative stress, and expression of PPAR receptors and the hydrolytic enzyme ASAHL/NAAA (N-acylethanolamine-hydrolyzing acid amidase). Results demonstrated that none of the NAEs showed cytotoxicity at tested concentrations. All three compounds significantly reduced pro-inflammatory markers (TNFα, CD86, CD68, IL1β, IL6, and P2RX7), though their efficacy profiles differed: DHEA was most effective against TNFα and IL1β, SDEA against CD86 and IL6, and EPEA against CD68 and P2RX7. Additionally, NAEs elevated the anti-inflammatory marker Arg-1 and CREB levels, suggesting a shift toward a neuroprotective microglial phenotype. In terms of antioxidant activity, EPEA and DHEA were most effective in suppressing lipid peroxidation (MDA) and reactive oxygen species (ROS), while all NAEs moderately reduced nitric oxide (NO) production. Furthermore, NAEs upregulated PPAR-α (strongest induction by EPEA) and PPAR-γ (maximal activation by SDEA), suggesting the involvement of nuclear receptor pathways in their mechanisms. Notably, SDEA and EPEA markedly increased ASAHL/NAAA expression, indicating their accelerated hydrolysis and potential metabolic conversion. These findings highlight the structure-dependent bioactivity of NAEs, with longer-chain, highly unsaturated DHEA showing prolonged effects, while SDEA and EPEA exhibited potent but shorter-lived actions. The data support further exploration of NAEs as targeted therapeutics for neuroinflammatory and neurodegenerative disorders.</p>