Effects of Electroacupuncture via AFG3L2 on Learning and Memory in Mice with Alzheimer's Disease and the Underlying Mechanisms

ObjectiveTo observe the effects of electroacupuncture (EA) at the Baihui (DU20) and Shenting (DU24) acupoints on learning and memory function in Alzheimer's disease (AD) mice, and to explore the underlying mechanisms.MethodsFour male 5×familial Alzheimer's disease (5×FAD) and four female wild-type (WT) mice, all aged two months, were selected and cross-bred. After approximately one month, their offspring were genotyped. Twenty 3-month-old offspring identified as positive were randomly assigned using a random number table to either a control group or an electroacupuncture (EA) group, with 10 mice in each group. Additionally, ten 3-month-old negative littermates were selected as the WT group. Following one week of adaptive fixation, EA group was administered at the Baihui(DU20) and Shenting(DU24) acupoints with a needle depth of 2 mm, using a sparse waveform at a frequency of 1/20 Hz for 30 minutes daily, five times a week, over a period of four weeks. WT and control group mice were captured and fixed under the same conditions without receiving any other treatment. Employing novel location recognition test (total distance travelled, discrimination index) to evaluate learning and memory function in mice; Chemically Exchange Saturated Transfer (CEST) imaging scans were performed using a 7. 0 T small animal magnetic resonance imaging system to detect glucose metabolism levels (asymmetric magnetisation transfer rate [MTRasym(Δω)]) in the hippocampus of mice. Immunohistochemistry was used to detect the level of Aβ-positive area in the hippocampus of mice. ATP assay kit was used to detect the concentration of ATP in the hippocampus of mice. Immunofluorescence was used to detect the positive area of Essential Mitochondrial Calcium Uniporter Regulator(EMER) in the hippocampus of mice. Western blot was used to detect the expression levels of mitochondrial calcium uniporter (MCU) and AFG3 like matrix AAA peptidase 2 (AFG3L2) proteins in the hippocampus of mice.Results(1) Novel location recognition test：On days 2 and 3 post-intervention, the total distance travelled across all three activity groups showed no statistically significant difference (P>0.05). Compared with the WT group, the control group exhibited a significantly reduced discrimination index (P