Electroacupuncture at Fengchi (GB20) and Gongxue (Extra) could promote dendritic spine formation, enhance synaptic plasticity, and improve cognitive function in rats with chronic sleep deprivation, and these effects may be related to the inhibition of the PI3K/AKT/mTOR signaling pathway and activation of hippocampal neuronal autophagy.
Key Findings
Results
Chronic sleep deprivation (CSD) induced by the modified multiple-platform water environment method impaired cognitive function in rats, as evidenced by prolonged escape latency and reduced platform crossings in the Morris water maze.
Model group rats showed prolonged escape latency and fewer platform crossings compared to the control group (P<0.05).
65 Wistar rats with normal cognitive function were screened; 12 served as controls and the remaining underwent CSD modeling, with 48 successfully modeled rats used in subsequent groups.
The CSD model was induced using a modified multiple-platform water environment method.
Activator group (treated with PI3K activator 740Y-P) also showed prolonged escape latency and fewer platform crossings compared to control (P<0.05), similar to the model group.
Results
CSD caused structural deterioration of hippocampal CA1 pyramidal neurons, including reduced dendritic branching and lower dendritic spine density.
Model group and activator group rats exhibited fewer dendritic branches and lower spine density in hippocampal CA1 pyramidal neurons compared to the control group (P<0.05).
Dendritic morphology and spine density were assessed using Golgi staining.
These structural changes indicate impaired synaptic plasticity in CSD rats.
Results
CSD upregulated the PI3K/AKT/mTOR signaling pathway in hippocampal tissue, as indicated by increased phosphorylation ratios of PI3K, AKT, and mTOR.
Compared to the control group, the model group and activator group showed increased ratios of p-PI3K/PI3K, p-AKT/AKT, and p-mTOR/mTOR (P<0.05).
Protein expression was measured by Western blot in hippocampal tissues.
Activation of PI3K/AKT/mTOR was associated with suppressed neuronal autophagy.
Results
CSD suppressed hippocampal neuronal autophagy, as shown by decreased LC3II/LC3I ratio, reduced Beclin-1 protein expression, and decreased positive expression of Beclin-1 and LC3 in the CA1 region.
Model group and activator group showed decreased LC3II/LC3I ratios and decreased Beclin-1 expression compared to controls (P<0.05).
Positive expression of Beclin-1 and LC3 in hippocampal CA1 region was also reduced in the model and activator groups (P<0.05).
Autophagy markers were assessed by Western blot and immunofluorescence staining.
The PI3K activator 740Y-P (0.02 mg/kg, intraperitoneal injection) replicated the autophagy-suppressing effects seen in the model group, confirming the role of PI3K/AKT/mTOR activation in autophagy suppression.
Results
Electroacupuncture (EA) at bilateral Fengchi (GB20) and Gongxue (Extra) improved cognitive function in CSD rats, shortening escape latency and increasing platform crossings.
Compared to the model group, activator group, and EA+activator group, the EA group showed shortened escape latency and increased platform crossings (P<0.05).
EA was administered for 20 minutes per session, once daily for 14 days.
The EA+activator group (which received both EA and the PI3K activator 740Y-P) did not show the same cognitive improvements as the EA-alone group, suggesting the PI3K/AKT/mTOR pathway mediates EA's beneficial effects.
Results
EA treatment enriched dendritic branching and increased spine density in hippocampal CA1 pyramidal neurons of CSD rats.
Compared to the model group, activator group, and EA+activator group, EA group rats showed enriched dendritic branching and higher spine density (P<0.05).
Structural improvements were assessed by Golgi staining of the hippocampal CA1 region.
These findings indicate EA enhances synaptic plasticity in CSD-affected hippocampal neurons.
Results
EA inhibited the PI3K/AKT/mTOR signaling pathway in CSD rats, decreasing phosphorylation ratios of PI3K, AKT, and mTOR in hippocampal tissue.
Compared to the model group, activator group, and EA+activator group, the EA group showed decreased ratios of p-PI3K/PI3K, p-AKT/AKT, and p-mTOR/mTOR (P<0.05).
The PI3K activator 740Y-P counteracted EA's inhibitory effects on this pathway when co-administered (EA+activator group).
Results were measured by Western blot in hippocampal tissues after 14 days of intervention.
Results
EA activated hippocampal neuronal autophagy in CSD rats, elevating LC3II/LC3I ratio, Beclin-1 protein expression, and positive expression of Beclin-1 and LC3 in the CA1 region.
Compared to the model group, activator group, and EA+activator group, the EA group showed elevated LC3II/LC3I ratios and increased Beclin-1 protein expression (P<0.05).
Positive expression of Beclin-1 and LC3 in hippocampal CA1 was increased in the EA group (P<0.05) compared to the model, activator, and EA+activator groups.
The co-administration of PI3K activator 740Y-P blocked EA's autophagy-enhancing effects, suggesting EA promotes autophagy via PI3K/AKT/mTOR pathway inhibition.
Autophagy markers were assessed by both Western blot and immunofluorescence staining.
What This Means
This research suggests that chronic sleep deprivation causes measurable damage to memory and learning in rats, accompanied by structural deterioration of brain cells in the hippocampus (a brain region critical for memory) and suppression of a cellular recycling process called autophagy. The study found that this damage is linked to overactivation of a specific molecular signaling chain (PI3K/AKT/mTOR), which normally helps cells grow but, when overactive, appears to shut down the cellular cleanup process that neurons need to stay healthy.
The study found that a form of acupuncture using mild electrical stimulation (electroacupuncture) at two specific points—Fengchi (GB20) and Gongxue (Extra)—performed once daily for 14 days reversed many of these harmful effects. Treated rats performed significantly better in a water maze memory test, showed richer branching and denser spines on their hippocampal neurons (structural signs of better brain connectivity), and had restored levels of autophagy proteins. Crucially, when the PI3K pathway was artificially reactivated using a drug while the rats also received electroacupuncture, the benefits largely disappeared—providing evidence that the PI3K/AKT/mTOR pathway is a key mechanism through which the treatment works.
This research suggests that electroacupuncture may help protect brain function during chronic sleep deprivation by rebalancing a molecular pathway that governs cellular maintenance in neurons. While this is an animal study and the findings cannot be directly applied to humans without further research, it provides a possible biological explanation for how acupuncture at these specific points might benefit cognition in sleep-deprived individuals.
Wu J, Cai P, Zhang S, Lu G, Guo C, Jia K, et al.. (2026). [Effect of electroacupuncture on neuronal autophagy in rats with chronic sleep deprivation via the PI3K/AKT/mTOR signaling pathway].. Zhongguo zhen jiu = Chinese acupuncture & moxibustion. https://doi.org/10.13703/j.0255-2930.20241130-k0002