AMPK mediates the anti-ferroptosis effect of acupuncture in cerebral ischemia–reperfusion rats

Background

Acupuncture is clinically effective in the treatment of ischemic stroke. The report suggests that energy stress inhibits ferroptosis in part through AMPK. However, whether the effect of activating AMPK is related to ferroptosis in ischemic stroke and whether acupuncture can achieve neuroprotection against ischemic brain injury through the pathway of inhibiting ferroptosis by activating AMPK has not been confirmed.

Methods

In this experiment, all rats were randomly divided into 4 groups: Sham group, MCAO/R group, MA (MCAO/R + acupuncture) group, and MAM (MCAO/R + acupuncture + metformin) group. The middle cerebral artery occlusion/reperfusion injury (MCAO/R) model was created by the wire embolism method, and the MA and MAM groups received acupuncture treatment with electrotherapy (1 mA, 2/15 Hz, 20 min each), while the MAM group continued to receive metformin (oral gavage 200 mg/kg) after successful modelling. Neurological deficit score and infarct volume were measured, Prussian blue staining and mitochondrial structural changes were observed, and Fe2+ and MDA levels were determined in the brain tissue of the rats. Western blot results were analyzed to determine differences in the expression of TFR1, SLC7A11, GPX4 and AMPK、p-AMPK proteins in order to explore the possible pathological processes involved in cerebral ischemia at behavioral, histological and molecular levels and the possible protective mechanisms of acupuncture.

Results

Acupuncture attenuated ischemia–reperfusion-induced brain damage and mitochondrial damage. Further studies showed that acupuncture reduced the levels of Fe2+, MDA and the expression of TFR1 protein and increased the expression of SLC7A11 and GPX4 protein in the diseased hippocampal region of MCAO/R rats. In addition, metformin, as an AMPK activator, significantly enhanced the protective effect of acupuncture on cerebral ischemic injury and enhanced the acupuncture-mediated reduction of Fe2+, MDA levels and TFR1 protein expression and the increase of SLC7A11 and GPX4 protein expression in the lesioned hippocampal region of MCAO/R rats.

Conclusions

These findings suggest that acupuncture can inhibit ferroptosis and thus exert a protective effect against ischemic brain injury, and that this mechanism may be achieved by activating AMPK. This extends the mechanism of action of acupuncture in the treatment of ischemic stroke.