温阳复元方抑制脑缺血再灌注大鼠神经元坏死性凋亡和线粒体分裂OACHSSCD

Objective:To elucidate the mechanisms of the Wenyang Fuyuan Formula in treating cerebral ischemia-reperfusion inju-ry(CIRI)in rats by detecting indicators related to the receptor-interacting protein kinase 1(RIPK1)/mixed lineage kinase domain-like protein(MLKL)/phosphoglycerate mutase 5(PGAM5)signaling pathway.Methods:Rats were randomly divided into a sham-operation group,model group,necrostatin-1(Nec-1)group,and Wenyang Fuyuan Formula group using a random number table.A rat CIRI model was established using the filament occlusion method.After one week of intervention with Wenyang Fuyuan Formula or Nec-1,2,3,5-triphenyltetrazolium chloride(TTC)staining was performed to determine successful model establishment.Neuro-logical deficits were evaluated using the Bederson scoring system.Hematoxylin-eosin(HE)staining and Nissl staining were used to observe the extent of brain tissue damage.Immunofluorescence was used to detect the expression of RIPK1 and PGAM5.Real-time quantitative reverse transcription polymerase chain reaction(RT-qPCR)and Western blot(WB)were used to detect the mRNA and protein expression levels of RIPK1,RIPK3,MLKL,PGAM5,and Drp1.Results:Compared with the sham-operation group,rats in the model group exhibited obvious white infarct areas in brain tissue,increased Bederson scores(P＜0.05),and severe brain tissue damage.Nissl staining showed increased neuronal apoptosis(P＜0.05),and the mRNA and protein expression levels of RIPK1,RIPK3,MLKL,PGAM5,and Drp1 in brain tissue were significantly increased(P＜0.05).Compared with the model group,rats in the Nec-1 group and the Wenyang Fuyuan Formula group showed smaller infarct areas,lower Bederson scores(P＜0.05),more Nissl bodies,and reduced neuronal apoptosis in brain tissue.The mRNA and protein expression levels of RIPK1,RIPK3,MLKL,PGAM5,and Drp1 were significantly decreased(P＜0.05),with a more pronounced reduction in the Wenyang Fuyuan Formula group.Conclusion:The Wenyang Fuyuan Formula may exert neuroprotective effects by downregulating the expression of proteins related to the RIPK1/MLKL/PGAM5 signaling pathway in CIRI rats,reducing mitochondrial fission,and further inhibiting neuronal necroptosis,thereby alleviating cellular damage.