基于串联质谱标签的电针改善阿尔茨海默病大鼠学习记忆损伤的海马组织蛋白质组学研究OA

Objective To investigate the effects of electroacupuncture(EA)on learning and memory abilities and hippocampal tissue proteome in Alzheimer's disease(AD)rats,and to explore differential proteins and regulatory pathways involved in the prevention and treatment of AD by EA.Methods Twenty-four SD rats were randomly divided into sham operation(Sham)group,Model group,and EA group,with 8 rats in each group.The AD model was established through injection of amyloid β protein 1-42 into bilateral hippocampal CA1 region.Following 7-week EA intervention,the learning and memory abilities of rats were evaluated using the Morris water maze experiment,and the recognition memory and novelty preference abilities of rats were assessed through the novel object recognition experiment.Hippocampal proteomic profiling was performed through tandem mass tag-labeling quantitative proteomics.Differential proteins underwent Gene Ontology(GO)and Kyoto Encyclopedia of Genes and Genomes(KEGG)enrichment analyses.The key differential proteins were verified using the parallel reaction monitoring(PRM)method.Results Compared with the Sham group,the rats in the Model group exhibited significantly prolonged escape latency(P＜0.01),reduced platform crossings numbers(P＜0.01),and decreased target quadrant dwell time(P＜0.01).Compared with the Model group,the escape latency of rats in the EA group was significantly shorter than that of the Model group(P＜0.01),the number of platform crossings was significantly increased(P＜0.01),and the target quadrant dwell time was significantly prolonged(P＜0.01).There was no significant difference in swimming speed among the different groups of rats(P＞0.05).During the exploration of the same objects,there was no significant difference in the recognition index of the same objects among all groups of rats(P＞0.05);during the exploration of new objects,compared with the Sham group,the recognition index of new objects in the Model group was significantly decreased(P＜0.01),and compared with the Model group,the recognition index of novel objects in the EA group was significantly increased(P＜0.01).The proteomic analysis results showed that a total of 211 differentially expressed proteins were identified.Based on these differences,11 significant reversal target proteins were selected.KEGG enrichment results highlighted synaptic vesicle cycle and glutamatergic synapse pathways as primary regulatory mechanisms.Two key proteins were finally identified,namely protein tyrosine phosphatase non-receptor type 5(PTPN5)as well as sodium and chloride-dependent γ-aminobutyric acid transporter 3(GAT3),which were associated with the glutamatergic synaptic pathway and the synaptic vesicle pathway respectively.The PRM results indicated that compared with the Sham group,the expressions of PTPN5 and GAT3 proteins in the hippocampal tissue of the Model group were increased(P＜0.01);compared with the Model group,the expressions of PTPN5 and GAT3 proteins in the hippocampal tissue of the EA group were decreased(P＜0.01),which was consistent with the proteinomics trends.Conclusions EA may improve the learning and memory abilities of AD rats by regulating the expressions of PTPN5 and GAT3 in the hippocampal tissue.