Electroacupuncture pretreatment alleviates cerebral Ischemia-Reperfusion-Induced intestinal barrier injury and neuroinflammation via inhibition of the TLR4/NLRP3 inflammasome pathway in rats

Abstract

This study investigates the protective effects and mechanisms of electroacupuncture (EA) pretreatment on intestinal barrier dysfunction following cerebral ischemia–reperfusion injury (CIRI), with a focus on the TLR4/NLRP3 inflammasome pathway. A middle cerebral artery occlusion (MCAO) rat model was established, with groups divided into Sham, ischemia–reperfusion (I/R), and EA pretreatment (I/R + EA). Neurological function and intestinal pathology were dynamically assessed at 2, 24, and 72 h post-reperfusion. Results demonstrated that EA pretreatment significantly attenuated CIRI-induced intestinal mucosal edema, inflammatory infiltration, and fibrosis, while improving neurological deficits. Mechanistically, EA pretreatment suppressed the TLR4/NF-κB/NLRP3 inflammasome axis, reducing pro-inflammatory cytokines (IL-1β, IL-18) and enhancing anti-inflammatory factors (TGF-β1, IL-4) and the antioxidant protein TRX1. Concurrently, it lowered plasma levels of lipopolysaccharide (LPS), diamine oxidase (DAO), and D-lactate, restoring intestinal epithelial tight junctions. 16S rDNA sequencing revealed that EA pretreatment reversed CIRI-induced gut microbiota dysbiosis, inhibiting pathogenic Escherichia-Shigella proliferation and rebalancing the Firmicutes/Bacteroidetes ratio. This study firstly identifies that EA pretreatment establishes a “neuro-immune-microbial” protective network through spatiotemporal modulation of the TLR4/NLRP3 pathway and microbiota interactions, providing a novel theoretical foundation for the “brain-gut co-protection” strategy in ischemic stroke therapy.