Yuan Yingxue, Zhang Junhong, Zhao Jingyi, Zhang Xiru, Cao Zhixin
Shandong Provincial Hospital Affiliated to Shandong First Medical University, Department of Pathology, Shandong, China.
Shandong Provincial Hospital Affiliated to Shandong First Medical University, Department of Otolaryngology-Head and Neck Surgery, Shandong, China.
Braz J Otorhinolaryngol. 2025 Jul 9;91(6):101674. doi: 10.1016/j.bjorl.2025.101674.
Screen for differential proteins in the cochlea of mice associated with Age-Related Hearing Loss (ARHL), analyze and validate the expression of specific differential proteins and genes in the cochlea and auditory cortex of ARHL mice, and preliminarily explore their potential mechanisms of action.
ABR (Auditory Brainstem Response) hearing tests were conducted to select 15-month-old C57BL/6 mice with significantly decreased hearing as the experimental group and 2-month-old mice with normal hearing as the control group. Cochleae were dissected, and unlabeled quantitative proteomics was employed to identify and analyze differentially expressed proteins in the inner ear of the two groups of mice. Key node proteins were selected via the STRING database and Cytoscape analysis. The expression of two selected proteins, ApoE and Spp1, in the cochlea was detected using qRT-PCR, Western blot, and immunofluorescence techniques, and their expression in the auditory cortex of the brain was further explored.
Label-free quantitative proteomics identified 115 differentially expressed proteins in the cochlea of 15-month-old ARHL mice compared to 2-month-old hearing-normal mice, including 42 upregulated and 73 downregulated proteins. GO and KEGG enrichment analyses revealed significant enrichment of differentially expressed proteins in functions and signaling pathways associated with neurodegenerative diseases and neurotransmission. Protein-Protein Interaction (PPI) analysis using the STRING database and Cytoscape selection identified ApoE and Spp1 as key hub proteins. Validation via qRT-PCR, Western blot, and immunofluorescence demonstrated that both ApoE and Spp1 were highly expressed in the cochlea and auditory cortex of the ARHL mice compare to 2-month-old hearing-normal mice.
ApoE and Spp1 are upregulated in the cochlea of ARHL mice, particularly in spiral ganglion neurons, and in the auditory cortex, suggesting their potential involvement in the pathogenesis and progression of ARHL through the modulation of auditory neural conduction systems.
Level 2.
筛选与年龄相关性听力损失（ARHL）相关的小鼠耳蜗差异蛋白，分析并验证ARHL小鼠耳蜗及听觉皮层中特定差异蛋白和基因的表达，初步探究其潜在作用机制。
进行听性脑干反应（ABR）听力测试，选择听力显著下降的15月龄C57BL/6小鼠作为实验组，听力正常的2月龄小鼠作为对照组。解剖耳蜗，采用无标记定量蛋白质组学方法鉴定和分析两组小鼠内耳中差异表达的蛋白质。通过STRING数据库和Cytoscape分析选择关键节点蛋白。使用qRT-PCR、蛋白质免疫印迹和免疫荧光技术检测所选两种蛋白ApoE和Spp1在耳蜗中的表达，并进一步探究它们在大脑听觉皮层中的表达。
无标记定量蛋白质组学鉴定出15月龄ARHL小鼠耳蜗中与2月龄听力正常小鼠相比有115种差异表达蛋白，其中42种上调，73种下调。基因本体（GO）和京都基因与基因组百科全书（KEGG）富集分析显示，差异表达蛋白在与神经退行性疾病和神经传递相关的功能和信号通路中显著富集。使用STRING数据库和Cytoscape选择进行蛋白质-蛋白质相互作用（PPI）分析，确定ApoE和Spp1为关键枢纽蛋白。通过qRT-PCR、蛋白质免疫印迹和免疫荧光验证表明，与2月龄听力正常小鼠相比，ApoE和Spp1在ARHL小鼠的耳蜗和听觉皮层中均高表达。
ApoE和Spp1在ARHL小鼠的耳蜗中上调，特别是在螺旋神经节神经元中，在听觉皮层中也上调，提示它们可能通过调节听觉神经传导系统参与ARHL的发病机制和进展。
2级。
