Objective To investigate the mechanism of Cong’er-Tong’qiao Agent (CTA) against age-related sensorineural apoptosis in the C57BL/6J mouse by regulating NEFM/MAPK signaling pathway. Methods 54 C57BL / 6J mice at 1 month of age were equally divided into 3 groups. 10 mice drinked tap water until 2 months after birth as young control group (2M control group); 22 mice drinked tap water until 7 months of age as presbycusis control group(7M control group); another 22 mice daily drinked the CTA until 7 months of age as CTA treating group (7M CTA group) . After the expiration of the animal experiments in each group, whole cochlear basilar membrane, 5 animals in each group were micro-dissected out, trimmed and mounted in glycerin on glass slides as flat surface preparations for inner and outer hair cells (IHCs and OHCs) morphological examination, while cochleae of 5 animals in each group were used for paraffin embedment and slice for morphological examination on cochlear spiral ganglion neurons(SGNs). From the7M control group and the 7M CTA group, the cochlear samples of 6 animals in each group were used for detecting the NEFM abundance values by proteomic techniques, and other cochlear samples of 6 animals in each group were used for detecting the MAPK14 expression level by Real time PCR (RT-PCR) technique, and then the data results were statistically analysed and compared, and their molecular signal transducting mechanism wrer analyzed using String and Kegg databases. Results By comparison of IHCs and OHCs at basal turn near hook region or SGNs at corresponding region in each group, it was shown that all IHCs and OHCs or SGNs were intact at2M control group, while almost all IHCs and OHCs or SGNs were missing at 7M control group, but the losses of the IHCs and OHCs or SGNs were significantly reduced at 7M CTA group(p<0.05).Proteomic research found that NEFM was one of the abundant enrichment proteins with significant differences and at the top in rank of expression intensity, wich its abundance values in the 7M CTA group were significantly increased by comparison with 7M control group (p<0.01),while RT-PCR research showed that cochlear MAPK14 expression levels in the 7M CTA group were significantly decreased by comparison with 7M control group? (p<0.01). NEFM was closely related to the MAPK14-mediated MAPK signaling pathway and SOD1-mediated the antioxidant signaling pathway, and further, they interacted with ROS/BCl-2/Bax/caspase and so on , wtich were apoptotic or anti-apoptotic factors, to form up-regulated or down--regulated signaling relationship.? Conclusion The CTA possessd the effect against age-related sensorineural apoptosis. The mechanism of its action may be that NEFM/MAPK14 regulated the SOD1/ROS-mediated signaling pathway of anti-apoptosis to achieve pharmacodynamic effect. It suggested that NEFM and MAPK14 may be one of the important targets of CTA for effectively resisting age-related sensorineural apoptosis.