Formononetin downregulates P53/SAT1/ACSL4 pathway-mediated ferroptosis to improve hypoxic-ischemic brain injury in neonatal mice
GUO Tao
CHEN Bolin
YANG Xiao
ZHAO Yanli
LI Xiaomin
HE Jiahao
SHI Jinsha
ZUO Hanjun
LI Juanjuan
Abstract：Objective To investigate the neuroprotective effects of formononetin(FMN)against hypoxic-ischemic brain damage(HIBD)in neonatal mice and the underlying mechanism.Methods Twenty-four neonatal C57BL/6J mice were randomly divided(n=6)into sham-operated group,HIBD model group,HIBD+FMN-L(50 mg/kg)group,and HIBD+FMN-H(100 mg/kg)group.Mouse models of HIBD were established by left common carotid artery ligation followed by hypoxia(92%N₂,8%O₂)for 40 min.FMN at the two doses was administered by intraperitoneal injection,and 3 days later,brain tissues from the cortical ischemic penumbra were collected for assessing expressions of ferroptosis-related proteins(P53,SAT1,and ACSL4)using Western blotting and immunofluorescence staining and for detecting the levels of Fe²⁺,superoxide,malondialdehyde(MDA),and glutathione(GSH).In cultured HT22 neurons with oxygen-glucose deprivation(OGD),the effects of 100 μmol/L FMN,10 μmol/L Nutlin-3(a P53 agonist),or their combination on expressions of ferroptosis proteins,intracellular Fe²⁺,reactive oxygen species(ROS),lipid peroxidation,GSH,mitochondrial membrane potential,and cell viability were evaluated.Results In the neonatal mouse models of HIBD,FMN treatment significantly suppressed the protein expression of P53,SAT1,and ACSL4,reduced Fe²⁺,ROS,and MDA levels and increased GSH content in the cortical ischemic penumbra.In HT22 neurons with OGD,FMN obviously alleviated OGD-induced ferroptosis as shown by lowered expressions of the key ferroptosis proteins,reduced Fe²⁺ accumulation and lipid peroxidation,and significant increases of GSH levels,mitochondrial membrane potential,and cell viability.Mechanistic experiments showed that activation of P53 signaling by Nutlin-3 markedly reversed the protective effects of FMN.Conclusion FMN produces neuro-protective effects against HIBD in neonatal mice by mitigating neuronal ferroptosis,primarily through downregulation of the P53/SAT1/ACSL4 signaling pathway.
Keywords：formononetinhypoxic-ischemic brain damageferroptosisP53SAT1ACSL4
Publication Date：2026-03-20
Online Publishing Date：2026-04-01(First online date of this platform, not the publication date of the document)
Pages：11( 604-614 )