Electroacupuncture ameliorates insomnia with anxiety and depression via modulating slow-wave oscillation, neurotransmitter pathways, and complement system: Insights from EEG and proteomics

While acupuncture is an established intervention for insomnia, the neurobiological mechanisms linking its modulation of brain activity to molecular-level changes remain poorly understood. This study addresses a critical gap by integrating multimodal data to uncover how electroacupuncture regulates oscillatory activity in specific brain regions and correlates these changes with proteomic alterations. This study employed a multimodal approach combining clinical scales, polysomnography with 26-channel brainwaves, and proteomic analysis to evaluate the effects of acupuncture. Electroacupuncture significantly improved subjective sleep quality and alleviated anxiety and depression symptoms ( p < 0.001). Objectively, it enhanced slow-wave activity (delta, theta, and alpha bands) in key regions such as the frontal and temporal lobes during NREM sleep, while suppressing high-frequency oscillations (beta and gamma bands). Proteomic analysis identified 90 differentially expressed proteins specifically associated with electroacupuncture treatment, including RRM2, ATP5F1E, IRS4, FN1, F5, IDE, UQCRQ, ATP6V0D1, and PPIF. These proteins were enriched in pathways critical to neurotransmitter metabolism (e.g., GABAergic and serotonergic synapses) and complement/coagulation cascades. Correlation analysis revealed significant associations between slow-wave EEG activity and proteins such as UQCRQ and ATP6V0D1 across multiple brain regions ( p < 0.05). Electroacupuncture may alleviate insomnia with anxiety and depression by modulating slow-wave synchronization and high-frequency oscillations. This neurophysiological regulation might be mediated through key proteins in signaling pathways linking neural, immune, and metabolic functions. Our study thus provides evidence for a multilevel mechanism supporting electroacupuncture as a multimodal therapy. • Electroacupuncture (EA) offers a promising therapeutic alternative for insomnia comorbid with anxiety and depression, yet its mechanisms have remained elusive. Our study bridges this critical knowledge gap by deploying an innovative multimodal framework that synergistically combines clinical assessment, high-density EEG, and proteomics. Our result supported that EA's significant clinical benefits—marked improvements in sleep quality and mood scores—are underpinned by distinct, objective neurophysiological and molecular signatures. • A key finding is EA's capacity to recalibrate brain activity by enhancing restorative slow-wave oscillations while suppressing high-frequency arousal rhythms in critical regions like the frontal lobe. • Proteomic analysis provides a groundbreaking molecular narrative, linking these EEG changes to the specific modulation of proteins involved in core pathways such as neurotransmitter metabolism (e.g., IDE) and mitochondrial energy production (e.g., ATP5F1E).