As one of the widely-used broad-spectrum anti-cancer drug, ototoxicity occurrence limited the usage of Cisplatin in clinical practice. At the same time, Eleutheroside E are found with kindny and never protection. However, the mechanism of the protection is remained unclear. In this study, 27 C57BL/6J mice were used and divided into Ctrl group with normal Saline injected, Cis and Cis + EE group with injection of Cis and Cis + EE peritoneally for Cisplatin-reduced ototoxicity modelling and intervention, then cochlears were dissected for proteomics. Network pharmacology to predict the target and biological mechanism of Eleutheroside E against cisplatin ototoxicity and Eleutheroside E might play an antagonistic role through the regulation of MAPK and pyrogenic signaling pathway. Compare with Cis group, Cis+EE group mice have a lower threshold value in ABR result and the group shows less cell pyroptosis than Cis group. Morphology results also indicated that Cis+EE group reduced space of spiral ganglion and the vascular structure was relatively clear than the Cis group. In addition, compared with Cis group, mRNA expression of NLRP3, GSDMD, ASC, Caspase-1, IL-1β and IL-18 genes in Cis+EE group decreased. The expression of p-JNK, p-ERK, P-P38, NF-κB P65, Cleaved Caspase-1, GSDMD was decreased, and the expression of IκB was increased. This study demonstrated that Eleutheroside E exerts an antagonistic effect on Cisplatin ototoxicity by down-regulating the activity of MAPK/NF-κB/NLRP3 signaling pathway, reducing the expression level of pyroptosis related proteins and inflammatory factors after cisplatin administration, and then inhibiting pyroptosis.