[Effects of electroacupuncture with different waveforms on NLRP3 inflammasome and pyroptosis in the cerebral cortex of rats with cerebral ischemia-reperfusion injury].

The MCAO/R model group showed significantly elevated neurological deficit scores, infarct volume, and markers of NLRP3 inflammasome-mediated pyroptosis compared to sham operation rats.

• Model group had significantly higher neurological deficit score and infarct volume percentage than sham operation group (P<0.01).
• Histological analysis revealed loose tissue structure, widened cytoplasmic gaps, and vacuolar degeneration in the model group.
• IL-1β and IL-18 levels were significantly higher in the model group than in the sham operation group (P<0.01).
• Protein and mRNA expression of NLRP3, Caspase-1, GSDMD, and ASC were all significantly higher in the model group than in the sham operation group (P<0.01).

All three electroacupuncture waveforms (continuous, discontinuous, and dense-disperse) significantly reduced neurological deficits and cerebral infarct volume compared to the model group.

• EA was performed at GV20 (Baihui), GV14 (Dazhui), and bilateral ST36 (Zusanli) once daily for 20 min for 7 days.
• All three EA groups showed reduced neurological deficit scores and infarct volume percentage compared to the model group (P<0.01).
• All three EA groups showed improved tissue damage on HE staining compared to the model group.
• IL-1β and IL-18 levels were decreased in all three EA groups compared to the model group (P<0.01).
• Protein and mRNA expression of NLRP3, Caspase-1, GSDMD, and ASC were lower in all three EA groups than in the model group (P<0.05, P<0.01).

The dense-disperse wave (EA-C, alternating 3 Hz/15 Hz) showed superior anti-pyroptotic effects compared to both continuous wave (EA-A) and discontinuous wave (EA-B) groups.

• Compared with the EA-A group, 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 and ASC mRNA expression (P<0.05, P<0.01).
• The EA-B (discontinuous wave, 15 Hz with 1.5 s intervals) group showed lower IL-1β level compared with the EA-A group (P<0.05), but fewer significant differences than the EA-C group.
• The dense-disperse wave used alternating frequencies of 3 Hz and 15 Hz, while the continuous wave used a fixed 15 Hz frequency.

Electroacupuncture treatment inhibited NLRP3 inflammasome components at both the protein and mRNA levels in the ischemic cerebral cortex.

• Western blot was used to detect protein expression of NLRP3, Caspase-1, GSDMD, and ASC.
• qRT-PCR was used to detect mRNA expression of NLRP3, Caspase-1, GSDMD, and ASC.
• All three EA waveforms reduced both protein and mRNA expression of NLRP3, Caspase-1, GSDMD, and ASC compared to the model group (P<0.05, P<0.01).
• The dense-disperse wave most effectively reduced ASC protein and Caspase-1 and ASC mRNA expression compared to both other waveforms.

A total of 70 male SD rats were used, with 48 successfully modeled MCAO/R rats divided into four groups of 12 for the intervention comparison.

• 12 rats were assigned to the sham operation group; the remaining 58 underwent MCAO/R modeling using the reformed Longa method.
• 48 successfully modeled rats were randomly divided into: model group, EA-continuous wave (EA-A) group, EA-discontinuous wave (EA-B) group, and EA-dense-disperse wave (EA-C) group, with 12 rats each.
• ELISA was used to measure IL-1β and IL-18 levels; TTC staining assessed cerebral infarct volume; HE staining observed morphological changes.
• Neurological deficit scores were recorded before and after intervention.