Macrophage Changes and High‐Throughput Sequencing in Aging Mouse Intervertebral Disks

ABSTRACT Background Intervertebral disk (IVD) degeneration is associated with lower back pain and aging; however, the mechanisms underlying age‐related changes and the changes in macrophage polarization in aging intervertebral disks require further elucidation. The aim of this study was to evaluate...

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Main Authors: Wei Wang, Cheng Jiang, Jiong‐Hui Chen, Yong‐Long Chen, Zhen‐Wu Zhang, Zhi‐Chao Yang, Jun Li, Xiao‐Chuan Li
Format: Article
Language:English
Published: Wiley 2025-06-01
Series:JOR Spine
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Online Access:https://doi.org/10.1002/jsp2.70061
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author Wei Wang
Cheng Jiang
Jiong‐Hui Chen
Yong‐Long Chen
Zhen‐Wu Zhang
Zhi‐Chao Yang
Jun Li
Xiao‐Chuan Li
author_facet Wei Wang
Cheng Jiang
Jiong‐Hui Chen
Yong‐Long Chen
Zhen‐Wu Zhang
Zhi‐Chao Yang
Jun Li
Xiao‐Chuan Li
author_sort Wei Wang
collection DOAJ
description ABSTRACT Background Intervertebral disk (IVD) degeneration is associated with lower back pain and aging; however, the mechanisms underlying age‐related changes and the changes in macrophage polarization in aging intervertebral disks require further elucidation. The aim of this study was to evaluate changes in macrophages, the differential expression of senescence genes, and their relationship with hub genes in IVDs during aging in mice. Methods Twenty‐eight male wild C57 mice aged 4 weeks were divided into two groups. Four mice per group were selected for high‐throughput sequencing and 10 for tail IVD immunohistochemical analysis. Adult and aged mouse IVD specimens were stained with hematoxylin–eosin, Fast Green, and Alcian Blue to determine collagen (Col) 1, Col2, proteoglycan, P16, P21, P53, CD11b, CD86, CD206, IL‐1, TGF‐β, and IL‐4 expression. High‐throughput sequencing was performed on adult and aged mouse IVD tissues. Results Aged mouse IVDs showed reduced height and marked degeneration, with decreased Col2 and proteoglycan expression and increased Col1 expression. The expression of senescence markers, senescence‐associated IL‐1, TGF‐β, and IL‐4, and macrophage‐related markers, CD11b, CD86, and CD206, increased markedly with age. High‐throughput sequencing revealed 1975 differentially expressed genes in adult and aged mice, with 797 genes showing upregulated expression (top five: Kcna7, Mmp9, Panx3, Myl10, and Bglap) and 1178 showing downregulated expression (top five: Srd5a2, Slc38a5, Gm47283, Npy, and Pcdh8). Gene Ontology and pathway enrichment analyses highlighted aging‐related cellular components, biological processes, and metabolic pathways. The identified hub genes included Cox5a, Ndufs6, and Ndufb9. Conclusions Disk senescence and reduced height in aged mice are linked to upregulated expression of senescence‐associated phenotypes and macrophage polarization markers. These findings suggest that macrophages and differential gene expression play key roles in age‐related IVD degeneration, indicating that they can be used as potential targets for therapeutic intervention.
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spelling doaj-art-14aa30f15d3c4c34a6fdb7ba9409292d2025-08-20T03:29:53ZengWileyJOR Spine2572-11432025-06-0182n/an/a10.1002/jsp2.70061Macrophage Changes and High‐Throughput Sequencing in Aging Mouse Intervertebral DisksWei Wang0Cheng Jiang1Jiong‐Hui Chen2Yong‐Long Chen3Zhen‐Wu Zhang4Zhi‐Chao Yang5Jun Li6Xiao‐Chuan Li7Chongqing Tongnan Hospital of Traditional Chinese Medicine Chongqing People's Republic of ChinaDepartment of Orthopaedic Surgery Gaozhou People's Hospital Gaozhou Guangdong Province People's Republic of ChinaDepartment of Orthopaedic Surgery Gaozhou People's Hospital Gaozhou Guangdong Province People's Republic of ChinaDepartment of Orthopaedic Surgery Gaozhou People's Hospital Gaozhou Guangdong Province People's Republic of ChinaDepartment of Orthopaedic Surgery Gaozhou People's Hospital Gaozhou Guangdong Province People's Republic of ChinaDepartment of Orthopaedic Surgery Gaozhou People's Hospital Gaozhou Guangdong Province People's Republic of ChinaDepartment of Orthopaedic Surgery Gaozhou People's Hospital Gaozhou Guangdong Province People's Republic of ChinaDepartment of Orthopaedic Surgery Gaozhou People's Hospital Gaozhou Guangdong Province People's Republic of ChinaABSTRACT Background Intervertebral disk (IVD) degeneration is associated with lower back pain and aging; however, the mechanisms underlying age‐related changes and the changes in macrophage polarization in aging intervertebral disks require further elucidation. The aim of this study was to evaluate changes in macrophages, the differential expression of senescence genes, and their relationship with hub genes in IVDs during aging in mice. Methods Twenty‐eight male wild C57 mice aged 4 weeks were divided into two groups. Four mice per group were selected for high‐throughput sequencing and 10 for tail IVD immunohistochemical analysis. Adult and aged mouse IVD specimens were stained with hematoxylin–eosin, Fast Green, and Alcian Blue to determine collagen (Col) 1, Col2, proteoglycan, P16, P21, P53, CD11b, CD86, CD206, IL‐1, TGF‐β, and IL‐4 expression. High‐throughput sequencing was performed on adult and aged mouse IVD tissues. Results Aged mouse IVDs showed reduced height and marked degeneration, with decreased Col2 and proteoglycan expression and increased Col1 expression. The expression of senescence markers, senescence‐associated IL‐1, TGF‐β, and IL‐4, and macrophage‐related markers, CD11b, CD86, and CD206, increased markedly with age. High‐throughput sequencing revealed 1975 differentially expressed genes in adult and aged mice, with 797 genes showing upregulated expression (top five: Kcna7, Mmp9, Panx3, Myl10, and Bglap) and 1178 showing downregulated expression (top five: Srd5a2, Slc38a5, Gm47283, Npy, and Pcdh8). Gene Ontology and pathway enrichment analyses highlighted aging‐related cellular components, biological processes, and metabolic pathways. The identified hub genes included Cox5a, Ndufs6, and Ndufb9. Conclusions Disk senescence and reduced height in aged mice are linked to upregulated expression of senescence‐associated phenotypes and macrophage polarization markers. These findings suggest that macrophages and differential gene expression play key roles in age‐related IVD degeneration, indicating that they can be used as potential targets for therapeutic intervention.https://doi.org/10.1002/jsp2.70061high‐throughput sequencingintervertebral diskmacrophagessenescence
spellingShingle Wei Wang
Cheng Jiang
Jiong‐Hui Chen
Yong‐Long Chen
Zhen‐Wu Zhang
Zhi‐Chao Yang
Jun Li
Xiao‐Chuan Li
Macrophage Changes and High‐Throughput Sequencing in Aging Mouse Intervertebral Disks
JOR Spine
high‐throughput sequencing
intervertebral disk
macrophages
senescence
title Macrophage Changes and High‐Throughput Sequencing in Aging Mouse Intervertebral Disks
title_full Macrophage Changes and High‐Throughput Sequencing in Aging Mouse Intervertebral Disks
title_fullStr Macrophage Changes and High‐Throughput Sequencing in Aging Mouse Intervertebral Disks
title_full_unstemmed Macrophage Changes and High‐Throughput Sequencing in Aging Mouse Intervertebral Disks
title_short Macrophage Changes and High‐Throughput Sequencing in Aging Mouse Intervertebral Disks
title_sort macrophage changes and high throughput sequencing in aging mouse intervertebral disks
topic high‐throughput sequencing
intervertebral disk
macrophages
senescence
url https://doi.org/10.1002/jsp2.70061
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