OBJECTIVES: To investigate the effect of acupuncture on the expressions of cuproptosis marker proteins-ferredoxin 1 (FDX1) and lipoic acid synthase (LIAS) in the ischemic penumbra (IP) of rats with cerebral ischemia-reperfusion injury (CIRI).
METHODS: SD rats were randomly divided into sham operation, model and acupuncture groups (=15). The CIRI model was established by occlusion of the middle cerebral artery and reperfusion (MCAO/R). In the acupuncture group, "Dazhui" (GV14), "Baihui" (GV20) and "Shuigou" (GV26) were punctured with filiform needles for 30 min, once every 12 h for 7 times. Neurological function was assessed using modified neurological severity scores (mNSS). Cerebral infarction volume was measured by TTC staining. Copper ion content in IP tissue was determined by colorimetric method. Mitochondrial respiratory chain complexes (MRCC)Ⅰ and Ⅱ contents were detected by ELISA. Protein expressions of FDX1, LIAS, and heat shock protein 70 (HSP70) were analyzed by Western blot. Mitochondrial morphology and structure in the IP region were observed via transmission electron microscopy (TEM). n
RESULTS: Compared with the sham operation group, the mNSS score (<0.01), the volume percentage of cerebral infarction (<0.000 1), copper ion in IP (<0.01), MRCCⅠ and Ⅱ contents (<0.000 1) and HSP70 protein expression (<0.001) were significantly increased, while the expression of FDX1 (<0.01) and LIAS (<0.001) were decreased in the model group. Compared with the model group, the mNSS score (<0.05), the volume percentage of cerebral infarction (<0.001), the contents of IP copper ion (<0.001), MRCCⅠ and Ⅱ contents (<0.01,<0.000 1) and HSP70 protein expression decreased (<0.01), whereas the protein expressions of FDX1 and LIAS were increased (<0.05) in the acupuncture group. TEM results showed that the outer membrane of mitochondria swelled and ruptured, the inner cristae ruptured and disappeared, and the density of the mitochondrial matrix decreased in the model group, which were relatively milder in the acupuncture group. P P P P P P P P P P P P P P
CONCLUSIONS: Acupuncture intervention can reduce the volume of cerebral infarction and improve the symptoms of neurological deficit in CIRI rats, and its mechanism may be related to reducing the loss of FDX1 and LIAS proteins and inhibiting the surge of HSP70 protein, thus preventing cuproptosis in brain cells.
