Holistic Regulation of Body-Mind Acupuncture Activates Leptin Receptor-AMPK Signaling to Restore Mitochondrial Homeostasis in Chronic Fatigue Syndrome in vitro and in vivo.

OBJECTIVE

To elucidate the therapeutic mechanism of holistic regulation of body and mind (HRBM) acupuncture in chronic fatigue syndrome (CFS) by focusing on leptin receptor (LepR)-AMP-activated protein kinase (AMPK) signaling to address mitochondrial dysfunction and oxidative stress.

METHODS

A CFS rat model was established by a 4-week treadmill-based exhaustive exercise protocol. After modeling, 50 rats were randomly allocated into 5 groups (n=10), control, CFS (model), acupuncture (Acu), Acu + Allo-aca, and Acu + Allo-aca + 5-aminoimidazole-4-carboxamide ribonucleoside (AICAR) groups. Rats in the Acu group received acupuncture at bilateral Baihui (GV 20) and Zusanli (ST 36) for 30 min daily over 4 weeks. Behavioral assessments were used to confirm model validity and treatment effects. Skeletal muscle transcriptomics were applied to identify differentially expressed genes associated with mitochondrial metabolism. LepR, AMPK, phosphorylated AMPK (p-AMPK), peroxisome proliferator-activated receptor γ coactivator 1α (PGC-1α), and nuclear factor erythroid 2-related factor 2 (Nrf2) were validated by RT-qPCR and Western blot analyses. In vitro, chronic fatigue-like cells were treated with EX-527 (1 µmol/L) and AICAR (500 µmol/L). Mitochondrial membrane potential was assessed using JC-1 assay, intracellular ATP and reactive oxygen species (ROS) levels were quantified, and expressions of apoptosis-related proteins, antioxidative enzymes, and mitophagy-associated proteins were evaluated by Western blot analysis. Mitochondrial ultrastructure was observed by transmission electron microscopy. Immunofluorescence and DCFH-DA probes were used to assess the LepR/AMPK co-localization and ROS dynamics.

RESULTS

Acupuncture at GV 20 and ST 36 significantly improved fatigue-related behaviors and exercise endurance in CFS rats. Compared with the control group, rats with CFS exhibited markedly aggravated fatigue symptoms (P<0.01), whereas acupuncture treatment significantly ameliorated these deficits (P=0.0015). Transcriptomic analysis revealed that acupuncture activated LepR-AMPK signaling pathway, accompanied by enhanced mitochondrial function, increased ATP production, and improved antioxidative capacity. Furthermore, acupuncture promoted PINK1/Parkin-mediated mitophagy and stabilized mitochondrial ultrastructure. In vitro, treatment with EX-527 and/or AICAR improved mitochondrial and metabolic measures in the chronic fatigue-like cell model.

CONCLUSIONS

HRBM acupuncture exerts therapeutic effects in CFS by activating LepR-AMPK signaling, thereby restoring mitochondrial bioenergetics, enhancing antioxidant defenses, and promoting mitophagy.