Protective effects and mechanisms of Huangzhu Qingnao granules against cerebral ischemia-reperfusion injury in rats
XUE Ruiwen
NIE Wei
XIONG Peng
WANG Dengkun
Abstract：Objective To investigate the protective effects of Huangzhu Qingnao granules(HZQNG)against cere-bral ischemia-reperfusion injury in rats subjected to middle cerebral artery occlusion/reperfusion(MCAO/R)and to ex-plore the underlying mechanism.Methods In vivo experiment,Wistar rats were randomly divided into the sham group,model group,butylphthalide group,and low-,medium-,and high-dose HZQNG groups,with 10 in each.Neurological deficits were evaluated using the Zea-Longa score.Histopathological changes,infarct area,blood-brain barrier(BBB)ul-trastructure,and apoptosis were assessed.Oxidative stress indices[reactive oxygen species(ROS),malondialdehyde(MDA),and superoxide dismutase(SOD)],serum inflammatory cytokines(IL-1β and IL-18),and the expression levels of thioredoxin-interacting protein(TXNIP),NOD-,LRR-and pyrin domain-containing protein 3(NLRP3),apoptosis-as-sociated speck-like protein containing a CARD(ASC),Caspase-1,and Gasdermin D(GSDMD)were measured.The in-teraction between TXNIP and NLRP3 was examined by co-immunoprecipitation.In vitro experiment,BV2 microglial cells were assigned into the normal group,model group,NLRP3 inhibitor group,and low-,medium-,and high-dose HZQNG groups.Cell viability was assessed using CCK-8.TXNIP-NLRP3 interaction was analyzed by co-immunoprecipitation,and the expression of TXNIP,NLRP3,ASC,Caspase-1 and GSDMD proteins was detected by Western blotting.Results In vivo experiment:The Zea-Longa neurological function score,the percentage of cerebral infarct area,the rate of neuronal apoptosis,the levels of ROS and MDA in the brain tissues,serum levels of IL-18 and IL-1β,as well as the protein expres-sion levels of TXNIP,NLRP3,ASC,Caspase-1,and GSDMD in the brain tissues,were as follows:model group>low-dose HZQNG group>medium-dose HZQNG group>butylphthalide group>high-dose HZQNG group>sham group(all P<0.05).In contrast,the SOD activity in the brain tissues followed the reverse order:sham group>high-dose HZQNG group>butylphthalide group>medium-dose HZQNG group>low-dose HZQNG group>model group(all P<0.05).HE staining results showed that the brain tissue structure in the sham group was essentially normal,while the model group ex-hibited disordered neuronal arrangement and significant structural damage.The pathological changes in the brain tissues were alleviated in the low-,medium-,and high-dose HZQNG groups,as well as in the butylphthalide group.Transmission electron microscopy revealed severe damage to the ultrastructure of the BBB in the cerebral cortex of the model group,whereas HZQNG intervention mitigated the extent of ultrastructural damage to the BBB.Co-immunoprecipitation confirmed the interaction between TXNIP and NLRP3 in rat brain tissues.Conclusion HZQNG ameliorates cerebral ischemia-re-perfusion injury in MCAO/R rats.The protective effects may be associated with alleviation of oxidative stress and inhibi-tion of the TXNIP/NLRP3/Caspase-1/GSDMD pathway-mediated inflammatory response and pyroptosis,thereby reducing BBB injury and promoting neurological recovery.
Keywords：Huangzhu Qingnao granulescerebral ischemia-reperfusion injurymiddle cerebral artery occlusion/reperfusionblood-brain barrierNLRP3 inflammasomepyroptosis
Publication Date：2026-02-25
Online Publishing Date：2026-03-19(First online date of this platform, not the publication date of the document)
Pages：7( 27-33 )