Geissoschizine methyl ether-(GME) is an indole alkaloid isolated from Uncariae Ramulus Cum Uncis, known for its various biological activities, including antioxidant effects, and neuroprotective properties. However, it has not yet been confirmed whether Geissoschizine methyl ether can protect dopaminergic neurons from death by inhibiting ferroptosis. In this study, First, network pharmacology is used to predict the action targets of GME in relation to Parkinson's disease and to perform pathway enrichment analysis. Molecular docking technology is then employed to further confirm the relevant action targets. Furthermore, we established a cell model using the ferroptosis inducers Erastin and MPP+ and found that Geissoschizine methyl ether exhibited an inhibitory effect on ferroptosis during the protection of dopaminergic neurons in Parkinson's disease. The JC-1 fluorescence probe kit was utilized to detect mitochondrial membrane potential; Mito-Tracker was used to analyze mitochondrial morphology and distribution; DCFH-DA was employed to detect mitochondrial ROS levels; and FerroOrange was used to assess the intracellular ferrous ion content. Western blotting was performed to measure the expression levels of GPX4 and SLC7A11 proteins in different treatment groups. The results of network pharmacology indicate that GME primarily exerts its effects by targeting AKT1, BCL2, DRD2, HTR2A, and other targets, with the Neuroactive ligand receptor interaction signaling pathway as the main pathway. Experimental results showed that Geissoschizine methyl ether improved the survival of MPP+-induced dopaminergic neurons by inhibiting ferroptosis. Therefore, Geissoschizine methyl ether is a potential drug for preventing the death of dopaminergic neurons by targeting and inhibiting ferroptosis.
