Zhongguo Zhen Jiu. 2026 Mar 12;46(3):393-401. doi: 10.13703/j.0255-2930.20241207-k0004. Epub 2025 Dec 25.
ABSTRACT
OBJECTIVE: To observe the effects of electroacupuncture (EA) with different waveforms on nucleotide-binding oligomerization domain-like receptor protein 3 (NLRP3) inflammasome and pyroptosis in the cerebral cortex of rats with cerebral ischemia-reperfusion injury (CIRI), and explore the potential mechanisms of EA in treating CIRI.
METHODS: A total of 70 male SD rats were selected. Twelve rats were randomly assigned to a sham operation group, while the remaining rats were subjected to middle cerebral artery occlusion/reperfusion (MCAO/R) model by reforming Longa method. Forty-eight successfully modeled rats were randomly divided into a model group, an EA-continuous wave (EA-A) group, an EA-discontinuous wave (EA-B) group, and an EA-dense-disperse wave (EA-C) group, with 12 rats in each group. EA was performed at "Baihui" (GV20), "Dazhui" (GV14), and bilateral "Zusanli" (ST36) once daily for 20 min for 7 d. The EA-A group used a continuous wave (15 Hz), the EA-B group used a discontinuous wave (15 Hz with 1.5 s intervals), and the EA-C group used a dense-disperse wave (alternating 3 Hz/15 Hz). Neurological deficit scores were recorded before and after intervention. After intervention, TTC staining was used to assess cerebral infarct volume. HE staining was performed to observe the morphological changes in the ischemic cortical brain tissue. ELISA was used to measure interleukin-1β (IL-1β) and interleukin-18 (IL-18) levels in the ischemic cortex. Western blot was used to detect the protein expression of NLRP3, Caspase-1, gasdermin D (GSDMD), and apoptosis-associated speck-like protein (ASC), and qRT-PCR was used to detect the mRNA expression of NLRP3, Caspase-1, GSDMD and ASC.
RESULTS: Compared with the sham operation group, the model group showed significantly higher neurological deficit score and infarct volume percentage (P<0.01); histological analysis revealed loose tissue structure, widened cytoplasmic gaps, and vacuolar degeneration in the model group. The IL-1β and IL-18 levels, and both protein and mRNA expression of NLRP3, Caspase-1, GSDMD, and ASC in the model group were significantly higher than those in the sham operation group (P<0.01). Compared with the model group, all three EA groups showed reduced neurological deficit scores and infarct volume percentage (P<0.01), improved tissue damage, and decreased levels of IL-1β and IL-18 (P<0.01). Protein and mRNA expression of NLRP3, Caspase-1, GSDMD, and ASC were also lower than those in the model group (P<0.05, P<0.01). Compared with the EA-A group, the EA-B group had lower IL-1β level (P<0.05), and the EA-C group had significantly lower levels of IL-1β and IL-18 (P<0.01, P<0.05), as well as decreased GSDMD and ASC protein expression and reduced Caspase-1 and ASC mRNA expression (P<0.05, P<0.01). Compared with the EA-B group, the EA-C group showed further decreases in ASC protein expression and Caspase-1, ASC mRNA expression (P<0.05, P<0.01).
CONCLUSION: Different EA waveforms could all alleviate neurological deficits and reduce cerebral infarct volume in CIRI rats. The mechanism may be related to inhibition of NLRP3 inflammasome-mediated pyroptosis.
PMID:41839600 | DOI:10.13703/j.0255-2930.20241207-k0004