Daily Nanosecond Pulsed Electric Field Brain Treatments Rescue D-gal-pretreated Mice from a SenescentAlzheimer-like Neuropathology with Spatial Exploration and Memory Decline

Background: Alzheimer’s disease (AD) remains without effective disease-modifying therapy, motivating the search for novel interventions that can counteract aging-related neuropathology. Method: We investigated whether daily administration of nanosecond pulsed electric fields (nsPEF) could reverse brain aging and AD-like pathology induced by chronic D-galactose (D-gal) exposure in mice. A senescent/AD-like phenotype was established through 12 weeks of daily intraperitoneal D-gal injections, followed by 14 consecutive days of nsPEF treatment. Results: D-gal administration induced pronounced neuropathology, including neuronal nuclear pyknosis, mitochondrial degeneration, amyloid-β42 (Aβ42) accumulation, and aberrant aggregation of Thr231-phosphorylated Tau (Thr231p-Tau). Senescence-associated markers (p16, p21), apoptotic mediators (caspase-3), and the pro-inflammatory cytokine TNF-α were significantly elevated. Fluoro-Jade C (FJC) staining confirmed widespread neuronal degeneration. Remarkably, nsPEF treatment initiated ultrastructural recovery within 6 hours, as evidenced by transmission electron microscopy. nsPEF restored nucleolar morphology, increased euchromatin content, reduced nuclear membrane infoldings, and rescued mitochondrial swelling and cristae loss. After two days, nsPEF normalized levels of p16, p21, and FJC-positive neurons, while markedly upregulating key nuclear-mitochondrial signaling proteins (SIRT1, HIF-1α, PGC-1α, Nrf1). After 14 days, Aβ42 deposits were cleared and the pathologically altered of Thr231p-Tau was corrected. Behavioral testing further demonstrated that nsPEF restored spatial exploration and memory functions mediated by the medial habenula in D-gal-treated mice. Importantly, identical nsPEF treatment applied to healthy mice did not induce neurodegeneration or alter caspase-3, PCNA, or TNF-α levels. Conclusions：Collectively, these results show that nsPEF treatment robustly reverses D-gal-induced senescent/AD-like neuropathology and rescues cognitive impairment. nsPEF may provide a safe, non-invasive, and mechanistically distinct strategy for preventing or treating of brain senescence, age-related AD, and potentially other neurodegenerative diseases.