Objective  "Astragalus-Leech" medicine pair can reduce cerebral ischemia-reperfusion injury (CIRI), but its mechanism of action is not yet clear. Ferroptosis is a new target of CIRI. In this paper, the mechanism of  the "Astragalus-Leech" medicine pair on regulating ferroptosis in the treatment of CIRI was investigated using the network pharmacology approach.

Methods  The active ingredients and targets of Astragalus-Leech were obtained by searching databases, such as PubChem, SwissTargetPrediction, Batman-TCM, UniProt, TCMSP and other databases, respectively; the CIRI-related targets were collected by searching GeneCards database; the Venny online tool was used to obtain the common targets of "Astragalus-Leech" medicine pairs for active ingredients and CIRI. Cytoscape software was used to construct a network of interrelationships between the active ingredients, disease and predicted targets of the "Astragalus-Leech" medicine pair, the protein interaction network was visualized, and CytoHubba plug-in was used to calculate the core targets. The GO analysis and KEGG analysis of the targets of "Astragalus-Leech" in the treatment of CIRI were performed using R language software. Using FerrDb database, the genes related to the regulation of ferroptosis were obtained, and the common genes among the active ingredients, CIRI and ferroptosis in the "Astragalus-Leech" medicine pair were analyzed to investigate their relationship and make predictions. AutoDockTools 1.5.7 and other softwares were used to verify the molecular docking between the active ingredients and key targets.

Results  Through searching the databases, 28 active ingredients of "Astragalus-Leech" medicine pair, 680 predicted gene targets of the drug pair, 1 513 targets related to CIRI, 253 common targets of drug pair-disease, 259 targets related to ferroptosis were obtained. 28 potential targets, including PIK3CA, RELA, MAPK1, MAPK8, PTGS2, STAT3, SRC, NOS2, etc. on the regulation of ferroptosis and intervention in CIRI, and 279 signaling pathways including PI3K-Akt, Ras, TNF, MAPK, and HIF-1 were obtained through related prediction. Molecular docking showed that there was an interaction between the key components of the drug pair and the core targets. The "Astragalus-Leech" medicine pair may intervene in the development of CIRI by regulating ferroptosis and exert its therapeutic effects.

Conclusion  Using network pharmacology methods, the potential targets and related pathways of "Astragalus-Leech" on the active ingredients by regulating ferroptosis against CIRI were analyzed, suggesting that "Astragalus-Leech" may play its role in anti-CIRI through oxidative stress and anti-inflammatory pathways to regulate ferroptosis pathway, and provide a basis for further cell and animal experiments.

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