温阳复元方通过内外源铁死亡途径改善脑缺血再灌注损伤的效应机制研究OA

Objective To investigate the mechanism of Wenyang Fuyuan Prescription(温阳复元方,WYFYP)by exploring its regula-tion of proteins related to endogenous and exogenous pathways to inhibit ferroptosis and exert its cerebral protective effects in a rat model of cerebral ischemia-reperfusion injury(CIRI).Methods Thirty-six healthy male SD rats were randomly divided into the sham-operated group,model group,Ferrostatin-1 group,and low-/medium-/high-dose WYFYP groups,with 6 rats in each group.Except for the sham-operated group,CIRI models were established in all other groups.Neurological deficit scores were assessed,and TTC staining was used to confirm the success of the rat modeling.HE staining and Nissl staining were used to compare brain tissue damage across groups,while immunofluorescence(IF)was employed to detect the protein expression of ACSL4 and NCOA4 in brain tissues.qRT-PCR was used to measure the relative mRNA expression levels of ACSL4,LPCAT3,NCOA4,and FTH1.Western blot(WB)was used to detect the relative protein expression levels of ACSL4,LPCAT3,NCOA4,FTH1,and GPX4.Results Compared with the sham-operated group,the model group showed large areas of white infarct foci(P<0.05),a significant increase in neurological deficit scores(P<0.05),loosely arranged brain cells,dissolved cytoplasm,fragmented nuclei,and severe loss of Nissl bodies on the infarcted side.The protein and mRNA expression contents of ACSL4,LPCAT3,and NCOA4 in the brain tissue were increased(P<0.05),while the pro-tein and mRNA expression of FTH1 and GPX4 were decreased(P<0.05).Compared with the model group,the Ferrostatin-1 group and the medium-dose WYFYP group showed a significant decrease in neurological deficit scores(P<0.05),reduced degree of brain tissue damage,and an increased number of surviving neurons.The medium-dose group also showed significantly decreased protein and mRNA expression of ACSL4,LPCAT3,and NCOA4(P<0.05),and significantly increased protein and mRNA expression of FTH1 and GPX4(P<0.05).Conclusion WYFYP improves neurological function and brain pathological brain tissue damage in CIRI rats,and its mecha-nism may be related to the inhibition of ferroptosis by regulating proteins associated with the ACSL4-LPCAT3 and NCOA4-FTH1 path-ways.