Cyclin-dependent kinase inhibitor-1 deficiency enhances bone destruction in a mouse model of arthritis

IF 3.1 Q3 ENDOCRINOLOGY & METABOLISM

Bone Reports Pub Date : 2025-12-01 DOI:10.1016/j.bonr.2025.101892

Kensuke Wada , Shinya Hayashi , Yuma Onoi , Shotaro Tachibana , Yoshihito Suda , Akira Saito , Takuma Maeda , Shotaro Araki , Kohei Motono , Tomoyuki Kamenaga , Masanori Tsubosaka , Yuichi Kuroda , Naoki Nakano , Tomoyuki Matsumoto , Ryosuke Kuroda

{"title":"Cyclin-dependent kinase inhibitor-1 deficiency enhances bone destruction in a mouse model of arthritis","authors":"Kensuke Wada , Shinya Hayashi , Yuma Onoi , Shotaro Tachibana , Yoshihito Suda , Akira Saito , Takuma Maeda , Shotaro Araki , Kohei Motono , Tomoyuki Kamenaga , Masanori Tsubosaka , Yuichi Kuroda , Naoki Nakano , Tomoyuki Matsumoto , Ryosuke Kuroda","doi":"10.1016/j.bonr.2025.101892","DOIUrl":null,"url":null,"abstract":"<div><div>Rheumatoid arthritis (RA) causes bone destruction by activating inflammatory cytokines and osteoclasts. Cyclin-dependent kinase inhibitor 1 (p21), a cell cycle regulator, may influence this process; however, its role remains unclear. Therefore, we investigated the effect and potential mechanisms of p21 deficiency in bone loss in a mouse model of arthritis. Collagen antibody-induced arthritis (CAIA) was established in p21 knockout (p21−/−) and wild-type mice. Bone destruction was analyzed using histology, micro-computed tomography, and bone strength testing; osteoclast formation and activity were evaluated using tartrate-resistant acid phosphatase (TRAP) staining and immunohistochemistry for cathepsin K. The expression of inflammatory cytokines and osteoclast-related genes was examined using immunohistochemistry and real-time polymerase chain reaction, respectively. p21−/− mice exhibited greater bone destruction and lower bone strength than wild-type mice. Additionally, TRAP and cathepsin K staining revealed significantly higher osteoclast count in p21−/− mice. Interleukin (IL)-6, IL-1β, tumor necrosis factor-alpha (TNF-α), and phosphorylated signal transducer and activator of transcription 3 (STAT3) levels were considerably higher in bone tissues of p21−/− mice than in those of wild-type mice. In vitro osteoclast differentiation in bone marrow macrophages (BMMs) was examined after IL-6 stimulation; osteoclast differentiation and osteoclast marker gene expression were significantly enhanced in p21−/− BMMs. Western blotting confirmed increased STAT3 phosphorylation in p21−/− BMMs; IL-6 treatment further amplified osteoclastogenesis in p21−/− BMMs. In conclusion, p21 deficiency exacerbates bone destruction in arthritis by promoting osteoclast differentiation and inflammatory cytokine expression via the IL-6/STAT3 pathway. Targeting p21 may offer therapeutic potential for preventing arthritis-related bone loss, such as in RA.</div></div>","PeriodicalId":9043,"journal":{"name":"Bone Reports","volume":"27 ","pages":"Article 101892"},"PeriodicalIF":3.1000,"publicationDate":"2025-12-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":"0","resultStr":null,"platform":"Semanticscholar","paperid":null,"PeriodicalName":"Bone Reports","FirstCategoryId":"1085","ListUrlMain":"https://www.sciencedirect.com/science/article/pii/S2352187225000695","RegionNum":0,"RegionCategory":null,"ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"Q3","JCRName":"ENDOCRINOLOGY & METABOLISM","Score":null,"Total":0}

引用次数: 0

Abstract

Rheumatoid arthritis (RA) causes bone destruction by activating inflammatory cytokines and osteoclasts. Cyclin-dependent kinase inhibitor 1 (p21), a cell cycle regulator, may influence this process; however, its role remains unclear. Therefore, we investigated the effect and potential mechanisms of p21 deficiency in bone loss in a mouse model of arthritis. Collagen antibody-induced arthritis (CAIA) was established in p21 knockout (p21^−/−) and wild-type mice. Bone destruction was analyzed using histology, micro-computed tomography, and bone strength testing; osteoclast formation and activity were evaluated using tartrate-resistant acid phosphatase (TRAP) staining and immunohistochemistry for cathepsin K. The expression of inflammatory cytokines and osteoclast-related genes was examined using immunohistochemistry and real-time polymerase chain reaction, respectively. p21⁻/⁻ mice exhibited greater bone destruction and lower bone strength than wild-type mice. Additionally, TRAP and cathepsin K staining revealed significantly higher osteoclast count in p21⁻/⁻ mice. Interleukin (IL)-6, IL-1β, tumor necrosis factor-alpha (TNF-α), and phosphorylated signal transducer and activator of transcription 3 (STAT3) levels were considerably higher in bone tissues of p21⁻/⁻ mice than in those of wild-type mice. In vitro osteoclast differentiation in bone marrow macrophages (BMMs) was examined after IL-6 stimulation; osteoclast differentiation and osteoclast marker gene expression were significantly enhanced in p21^−/− BMMs. Western blotting confirmed increased STAT3 phosphorylation in p21^−/− BMMs; IL-6 treatment further amplified osteoclastogenesis in p21^−/− BMMs. In conclusion, p21 deficiency exacerbates bone destruction in arthritis by promoting osteoclast differentiation and inflammatory cytokine expression via the IL-6/STAT3 pathway. Targeting p21 may offer therapeutic potential for preventing arthritis-related bone loss, such as in RA.

查看原文本刊更多论文

细胞周期蛋白依赖性激酶抑制剂-1缺乏增强关节炎小鼠模型的骨破坏

类风湿关节炎（RA）通过激活炎症细胞因子和破骨细胞导致骨破坏。细胞周期蛋白依赖性激酶抑制剂1 (p21)，一种细胞周期调节剂，可能影响这一过程；然而，它的作用仍不清楚。因此，我们在关节炎小鼠模型中研究了p21缺乏对骨质流失的影响及其潜在机制。在p21敲除（p21−/−）和野生型小鼠中建立胶原抗体诱导关节炎（CAIA）。采用组织学、显微计算机断层扫描和骨强度测试分析骨破坏情况；采用抗酒石酸酸性磷酸酶（TRAP）染色和组织蛋白酶k的免疫组化检测破骨细胞的形成和活性。分别采用免疫组化和实时聚合酶链反应检测炎症细胞因子和破骨细胞相关基因的表达。与野生型小鼠相比，P21−/−小鼠表现出更大的骨破坏和更低的骨强度。此外，TRAP和组织蛋白酶K染色显示p21−/−小鼠的破骨细胞计数显著增加。与野生型小鼠相比，p21−/−小鼠骨组织中白介素(IL)-6、IL-1β、肿瘤坏死因子-α (TNF-α)和磷酸化信号换能器和转录激活因子3 （STAT3）水平明显升高。IL-6刺激后观察骨髓巨噬细胞（BMMs）体外破骨细胞分化；破骨细胞分化和破骨细胞标记基因表达在p21−/−BMMs中显著增强。Western blotting证实p21−/−BMMs中STAT3磷酸化增加；IL-6治疗进一步放大了p21−/−BMMs的破骨细胞生成。综上所述，p21缺乏通过IL-6/STAT3通路促进破骨细胞分化和炎症细胞因子表达，从而加剧关节炎的骨破坏。靶向p21可能为预防关节炎相关的骨质流失提供治疗潜力，例如在RA中。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

求助全文

约1分钟内获得全文求助全文

来源期刊

Bone Reports Medicine-Orthopedics and Sports Medicine

CiteScore

4.30

自引率

4.00%

发文量

444

审稿时长

57 days

期刊介绍： Bone Reports is an interdisciplinary forum for the rapid publication of Original Research Articles and Case Reports across basic, translational and clinical aspects of bone and mineral metabolism. The journal publishes papers that are scientifically sound, with the peer review process focused principally on verifying sound methodologies, and correct data analysis and interpretation. We welcome studies either replicating or failing to replicate a previous study, and null findings. We fulfil a critical and current need to enhance research by publishing reproducibility studies and null findings.