To investigate a hippocampal anti-hypertensive mechanism induced by twirling reinforcing-reducing manipulation (TRRM) using proteomics in rats.

Forty-two male spontaneously hypertensive rats were randomly divided into 3 groups, and 14 Wistar-Kyoto rats were served as control group. In the twirling reinforcing (TRF) group, needles were directly inserted into the Taichong (LR 3) point, then thumbs were moved heavily forward and lightly backward for 3 min, while needles remained inserted for 17 min. In the twirling reduction (TRD) group, the same treatment was applied as in the TRF group except that the thumb moved lightly forward and heavily backward. In the model and control groups, only the corresponding grasping and fixation were applied. All interventions were conducted for 14 days. The blood pressure (BP) of all rats was measured one day before intervention and every other day after. Then hematoxylin-eosin (H&E) staining, label-free and parallel reaction monitoring proteomic techniques were used to assess hippocampal samples from each group.

Systolic BPs showed a significant decrease in the TRF and TRD groups compared with the model group (P < .01). In the model group, H&E staining showed obvious pathological changes in the hippocampus, while in the TRF and TRD groups, the hippocampal morphology was only slightly altered. Label-free proteomic analysis revealed 1163 differential protein expressions between groups. Gene Ontology enrichment analysis confirmed that the differentially expressed proteins were enriched in different biological processes, cellular components, and molecular functions. Kyoto Encyclopedia of Genes and Genomes enrichment analysis showed that TRRM proteins were expressed in the serotonergic synapse pathway, renin-angiotensin system, the mitogen-activated protein kinase signaling pathway, and the peroxisome pathway, which were all also related to BP regulation.

TRRM can significantly lower the BP of SHRs. The mode of action may be through the activation of various protein pathways in the hippocampus that are related to BP regulation.