Abstract: Objective: To identify and validate key interaction proteins of cobra cytotoxin-1（CTX-1） through network toxicology prediction combined with multi-dimensional experiments, construct a mouse skin necrosis model, and preliminarily investigate the association between CTX-1 binding to the identified proteins and skin necrosis by utilizing histopathological and immunohistochemical analyses. Methods: CTX-1-interacting proteins were initially predicted using the Super-PRED database. A Venn diagram identified overlapping proteins between these interactors and dermal necrosis-associated proteins from the GeneCards database. Protein-protein interaction（PPI） network analysis was performed using the STRING database, followed by mining of key proteins with Cytoscape software. One critical protein was selected for experimental validation. Molecular docking analyzed molecular interaction sites between CTX-1 and the selected key protein. Their interaction was validated in vivo and in vitro using bimolecular fluorescence complementation（BiFC） and surface plasmon resonance（SPR）. Mice were randomly divided into a control group and three experimental groups（n=8/group, half male/half female）. Mouse models of skin necrosis with concentration gradients of 40μg, 70μg and 100μg were constructed. Mouse skin tissues were collected for histopathological examination and immunohistochemistry（IHC）, and the expression of STAT3 in mouse skin tissues of each group was statistically analyzed. Results: The Venn diagram identified 48 potential CTX-1 interactors, with 37 forming a PPI network. Eleven key proteins were mined from these proteins, and STAT3 was selected for validation. Molecular docking revealed 11 hydrogen bonds formed between STAT3 and CTX-1 via 9 amino acids, which constituted their non-covalent interactions. Intracellular BiFC experiment preliminarily confirmed the interaction between CTX-1 and STAT3, while the in vitro SPR experiment yielded an affinity（KD） of 4.327×10^-8 M between CTX-1 and STAT3, demonstrating strong binding. HE-stained pathological sections showed significant necrosis and inflammatory cell infiltration in the skin tissues of mice in mild-and high-dose CTX-1 groups. IHC revealed higher STAT3-positive area ratios in all CTX-1 groups vs control groups, and the differences between mild-and high-dose CTX-1 groups and the control groups were statistically significant（P＜0.05）. Conclusion: Following cobra envenomation, CTX-1 in the venom may bind to STAT3 within the body, thereby triggering a cascade of pathophysiological reactions that contribute to local tissue swelling and necrosis.