中国耳鼻咽喉颅底外科杂志2025，Vol.31Issue(5)：6-12,7.DOI:10.11798/j.issn.1007-1520.202524543
聪耳通窍剂经NEFM/MAPK通路对抗小鼠老年性耳蜗感音神经细胞凋亡机制的探讨
Investigation on mechanism of Cong'er-Tong'qiao agent against age-related cochlear sensorineural apoptosis in mice via NEFM/MAPK pathway
摘要
Abstract
Objective To investigate the mechanism of Cong'er-Tong'qiao agent(CTA)against age-related cochlear sensorineural apoptosis in the C57BL/6J mice by regulating neurofilament medium polypeptide(NEFM)/mitogen-activated protein kinase(MAPK)signaling pathway.Methods Fifty-four C57BL/6J mice at 1 month of age were selected for the experiment.Among them,10 mice drank tap water until two months after birth as the young control group(2M control group).Twenty-two mice drank tap water until 7 months after birth as the control group of the age-related cochlear sensorineural apopt otic model(7M control group).Twenty-two mice were given the CTA until 7 months after birth as the CTA treating group(7 M CTA group).After the expiration of the animal experiments in each group,five animals in each group were used for whole cochlear basilar membrane spread preparations to observe the morphology of inner and outer hair cells,and five animals were used for cochlear epoxy resin embedding and sectioning to observe the morphology of spiral ganglion neurons(SGNs).In addition,from the 7M control group and the 7M CTA group,the cochlear samples of 6 animals were selected from each group to detect the abundance value of NEFM through proteomics technology,and the expression level of MAPK14 was detected in 6 animals' cochleae through real time PCR(RT-PCR)technology.The data were subjected to statistical analysis,and the molecular signal transduction mechanism was analyzed in combination with the String and Kegg databases.Results By comparison of inner and outer hair cells at basal turn near hook region or SGNs at corresponding region in each group,it was shown that all the inner and outer hair cells and SGNs were intact in the 2M control group,while almost all the inner and outer hair cells or SGNs were missing in the 7M control group,but the losses of the inner and outer hair cells and SGNs were significantly reduced in the 7M CTA group(P＜0.001).Proteomic studies revealed that NEFM,one of the abundant enrichment proteins with significant differences and at the top in rank of expression intensity,was significantly increased in abundance after intervention with CTA in the 7 M CTA group,compared to the 7M control group(P＜0.001).The RT-PCR study showed that,compared with the 7M control group,the expression level of cochlear MAPK14 in the 7M CTA group was significantly decreased(P＜0.001).The MAPK signaling pathway mediated by NEFM and MAPK14 was closely related to the antioxidant signaling pathway mediated by superoxide dismutase 1(SOD1),and subsequently formed an up-down signal transduction regulatory interrelationship with reactive oxygen species(ROS)/Bcl-2/Bax/caspase-9 and other factors related to apoptosis or anti-apoptosis.Conclusions The CTA has the effect of combating age-related sensorineural apoptosis.The mechanism may be that NEFM/MAPK14 regulate the SOD1/ROS-mediated signaling pathway of anti-apoptosis to achieve pharmacodynamic effect.It suggests that NEFM and MAPK14 may be one of the important targets of CTA on effectively resisting age-related sensorineural apoptosis.关键词
老年性聋/耳蜗/小鼠/神经丝中链多肽/丝裂原活化蛋白激酶14/中药Key words
Presbycusis/Cochlea/Mouse/NEFM/MAPK14/Traditional Chinese medicine分类
临床医学引用本文复制引用
宣伟军,文艺群,黄力毅,唐俊波,陈思仲,陆灵娟,丁大连..聪耳通窍剂经NEFM/MAPK通路对抗小鼠老年性耳蜗感音神经细胞凋亡机制的探讨[J].中国耳鼻咽喉颅底外科杂志,2025,31(5):6-12,7.基金项目
国家自然科学基金资助项目(81973913,81774374,81373700,81260552). （81973913,81774374,81373700,81260552）
