Zhongguo Zhong Yao Za Zhi. 2026 Apr;51(7):2029-2039. doi: 10.19540/j.cnki.cjcmm.20251217.801.
ABSTRACT
This study aimed to investigate whether Huangqi Chifeng Tang(HQCFT) attenuates neuronal injury and inflammatory responses induced by cerebral ischemia reperfusion injury(CIRI) through regulating the nuclear factor erythroid 2-related factor 2(Nrf2)/NOD-like receptor protein 3(NLRP3)/cysteinyl aspartate-specific proteinase-1(Caspase-1)/gasdermin D(GSDMD) pathway. Oxygen-glucose deprivation/reoxygenation(OGD/R) was employed to establish an in vitro model with BV2 murine microglial cells, while the middle cerebral artery occlusion/reperfusion(MCAO/R) model was induced in mice through the suture method. The cell experiments were conducted with control, model, si-NC, si-Nrf2, HQCFT, and si-Nrf2+HQCFT groups. The animal experiments encompassed the sham, model, ML385, HQCFT, and ML385+HQCFT groups. Cell viability and injury were assessed via CCK-8 and lactate dehydrogenase(LDH) assays. Neurological function and tissue damage were evaluated by the modified neurological severity score(mNSS), TTC staining, HE staining, and Nissl staining. Western blot was employed to determine the expression levels of pyroptosis-related proteins, including Nrf2, NLRP3, Cleaved Caspase-1, GSDMD-Nterminal domain(GSDMD-N), and Cleaved Caspase-3. RT-qPCR was performed to quantify the mRNA levels of Nrf2 and NLRP3. The release of pro-inflammatory cytokines, including tumor necrosis factor(TNF)-α, interleukin(IL)-1β, and IL-18, was assessed by ELISA. The results of cell experiments showed that HQCFT increased the viability of BV2 murine microglial cells following OGD/R injury(P<0.05), reduced LDH release(P<0.001), and suppressed the expression of pyroptosis-related molecules and the secretion of inflammatory cytokines. The results of animal experiments showed that HQCFT improved the neurological function(P<0.05), reduced the infarct volume(P<0.01), alleviated neuronal structural damage, reversed the downregulation of Nrf2 and the upregulation of NLRP3, Cleaved Caspase-1, GSDMD-N, and Cleaved Caspase-3 in the brain tissue(P<0.05), and decreased the serum levels of pro-inflammatory cytokines(P<0.05) in MCAO/R mice. Genetic silencing or pharmacological inhibition of Nrf2 gene exacerbated the injury and significantly attenuated the protective effects of HQCFT, indicating that its beneficial effects were partially dependent on Nrf2 gene activation. In conclusion, HQCFT exerts neuroprotective effects against CIRI, in part by activating the Nrf2 signaling pathway and thereby inhibiting both the NLRP3/Caspase-1/GSDMD canonical pyroptosis pathway and the Caspase-3-mediated pyroptosis pathway, ultimately alleviating neuroinflammatory responses.
PMID:42392783 | DOI:10.19540/j.cnki.cjcmm.20251217.801