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.