Shunaoxin dropping pills alleviate cerebral ischemia-reperfusion injury in MCAO rats associated with ferroptosis inhibition and AKT/Nrf2/HO-1 pathway activation

Ischemic stroke (IS), accounting for about 71% of stroke cases, is the most common form of this significant global cause of disability and mortality. Shunaoxin dropping pills (SNX), a traditional Chinese medicine, have demonstrated notable therapeutic benefits for IS patients. However, preclinical studies investigating its mechanisms remain scarce. This study examined the role of SNX in mitigating cerebral I/R injury by inhibiting Ferroptosis, with an emphasis on the Akt/Nrf2/HO-1 signaling pathway mechanisms. LC-MS/MS was used to analyze the chemical composition of SNX. Rats were administered SNX (45 or 90 mg/kg/d) for three consecutive days prior to MCAO model surgery. Neurological impairment in rats was evaluated and histopathological damage was assessed. The levels of Fe²⁺, MDA, and GSH in brain tissue were measured using commercial assay kits. The expression of GPX4, SLC7A11, ACSL4 were evaluated via western blot. The expression of FTH1 and FPN1 were detected by immunofluorescence. Network pharmacology identified potential SNX targets linked to Ferroptosis and ischemic stroke. The study then investigated the activation of the Akt/Nrf2/HO-1 signaling pathway by SNX. Following SNX treatment, MCAO rats exhibited notable enhancements in neurobehavioral scores, decreased infarct volume, and diminished pathological damage. SNX exerted neuroprotective effects by attenuating lipid peroxidation and inhibiting Ferroptosis. Network pharmacological analysis initially suggested, and subsequent experiments indicated, that the inhibition of Ferroptosis by SNX has a strong correlation with the activation of the Akt/Nrf2/HO-1 signaling pathway. In conclusion, SNX ameliorates cerebral I/R injury through reducing Ferroptosis, likely via Akt/Nrf2/HO-1 pathway activation.