Expression of ApoE and Spp1 in the cochlea and auditory cortex of age-related hearing loss mice

Objective: 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...

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Main Authors: Yingxue Yuan, Junhong Zhang, Jingyi Zhao, Xiru Zhang, Zhixin Cao
Format: Article
Language:English
Published: Elsevier 2025-11-01
Series:Brazilian Journal of Otorhinolaryngology
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Online Access:http://www.sciencedirect.com/science/article/pii/S180886942500117X
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author Yingxue Yuan
Junhong Zhang
Jingyi Zhao
Xiru Zhang
Zhixin Cao
author_facet Yingxue Yuan
Junhong Zhang
Jingyi Zhao
Xiru Zhang
Zhixin Cao
author_sort Yingxue Yuan
collection DOAJ
description Objective: 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. Methods: 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. Results: 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. Conclusion: 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 of evidence: Level 2.
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spelling doaj-art-5c0cce934e884121a16cd65d8b5e863f2025-08-20T02:36:16ZengElsevierBrazilian Journal of Otorhinolaryngology1808-86942025-11-0191610167410.1016/j.bjorl.2025.101674Expression of ApoE and Spp1 in the cochlea and auditory cortex of age-related hearing loss miceYingxue Yuan0Junhong Zhang1Jingyi Zhao2Xiru Zhang3Zhixin Cao4Shandong Provincial Hospital Affiliated to Shandong First Medical University, Department of Pathology, Shandong, ChinaShandong Provincial Hospital Affiliated to Shandong First Medical University, Department of Otolaryngology-Head and Neck Surgery, Shandong, ChinaShandong Provincial Hospital Affiliated to Shandong First Medical University, Department of Pathology, Shandong, ChinaShandong Provincial Hospital Affiliated to Shandong First Medical University, Department of Pathology, Shandong, ChinaShandong Provincial Hospital Affiliated to Shandong First Medical University, Department of Pathology, Shandong, China; Corresponding author.Objective: 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. Methods: 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. Results: 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. Conclusion: 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 of evidence: Level 2.http://www.sciencedirect.com/science/article/pii/S180886942500117XAge-related hearing lossApoESpp1Spiral ganglion neuronProteomics
spellingShingle Yingxue Yuan
Junhong Zhang
Jingyi Zhao
Xiru Zhang
Zhixin Cao
Expression of ApoE and Spp1 in the cochlea and auditory cortex of age-related hearing loss mice
Brazilian Journal of Otorhinolaryngology
Age-related hearing loss
ApoE
Spp1
Spiral ganglion neuron
Proteomics
title Expression of ApoE and Spp1 in the cochlea and auditory cortex of age-related hearing loss mice
title_full Expression of ApoE and Spp1 in the cochlea and auditory cortex of age-related hearing loss mice
title_fullStr Expression of ApoE and Spp1 in the cochlea and auditory cortex of age-related hearing loss mice
title_full_unstemmed Expression of ApoE and Spp1 in the cochlea and auditory cortex of age-related hearing loss mice
title_short Expression of ApoE and Spp1 in the cochlea and auditory cortex of age-related hearing loss mice
title_sort expression of apoe and spp1 in the cochlea and auditory cortex of age related hearing loss mice
topic Age-related hearing loss
ApoE
Spp1
Spiral ganglion neuron
Proteomics
url http://www.sciencedirect.com/science/article/pii/S180886942500117X
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AT jingyizhao expressionofapoeandspp1inthecochleaandauditorycortexofagerelatedhearinglossmice
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