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This study was conducted to investigate possible mechanisms underlying egg yolk phospholipids on improving memory using scopolamine–treated mice and PC12 cells. Oral administration of egg yolk phospholipids significantly decreased latency in passageway water maze tasks and reduced error hits in passive avoidance tests for scopolamine–treated mice (P < 0.05). As the results presented, egg yolk phospholipids attenuated spatial memory deficits via preventing region–specific lesions in hippocampus CA1-2 and regulating the activity of cholinergic biomarkers. Egg yolk phospholipids significantly inhibited scopolamine–induced increase of acetylcholinesterase activity and decrease of choline acetyltransferase activity (P < 0.05), thus maintaining the release of acetylcholine at a normal level. Further analysis showed that egg yolk phospholipids downregulated scopolamine–induced MAO activity and MDA level in PC12 cells (P < 0.05). These data suggest that egg yolk phospholipids might exert neuroprotective effects via attenuating cholinergic damage and inhibiting the oxidative stress in vitro.