压应力激活SOST/Wnt/β-catenin 通路诱导软骨终板细胞退变  

Compressive stress induces degeneration of cartilaginous endplate cells through the SOST/Wnt/beta-catenin pathway

作　　者：项攀 车艳军 罗宗平[1] Xiang Pan;Che Yanjun;Luo Zongping(Department of Orthopaedics,The First Affiliated Hospital of Soochow University,Suzhou 215006,Jiangsu Province,China;Orthopedics and Sports Medicine Center,The Affiliated Suzhou Hospital of Nanjing Medical University,Suzhou 215000,Jiangsu Province,China)

机构地区：[1]苏州大学附属第一医院骨科研究所,江苏省苏州市215006 [2]南京医科大学附属苏州医院,骨科与运动医学中心,江苏省苏州市215000

出　　处：《中国组织工程研究》2025年第5期951-957,共7页Chinese Journal of Tissue Engineering Research

基　　金：国家自然科学基金项目(32071307),项目负责人:罗宗平;江苏省高等学校重点学科建设项目(PAPD),项目负责人:罗宗平;苏州市科技局项目(SKY2022185),项目负责人:车艳军;姑苏卫生人才计划人才科研项目(GSWS2021035),项目负责人:车艳军;苏州市中西医结合科研基金(SKYD:2023254),项目负责人:车艳军。

摘　　要：背景:在许多可以导致椎间盘退变的因素中(衰老、营养匮乏、机械因素等),力学负荷被认为是极其重要的因素,但其机制尚不清楚。目的:探讨硬骨素和Wnt/β-catenin信号通路在压应力诱导终板软骨退变中的作用。方法:提取4周龄雄性SD大鼠软骨终板细胞,体外利用力学加载仪器对终板软骨细胞施加压应力,于压缩细胞1,3,5,7 d,采用CCK-8法测定细胞活力;Western blot、RT-qPCR及细胞免疫荧光等检查终板软骨细胞内软骨标记物(聚集蛋白聚糖、Ⅱ型胶原)、钙化相关因子(Runx2、骨钙素)、细胞外基质降解酶及信号通路基因(硬骨素、β-catenin)等表达。结果与结论:①压应力作用下,终板软骨细胞活力会随着压应力时间、强度增加而降低;②终板软骨细胞的软骨标记物(聚集蛋白聚糖、Ⅱ型胶原)表达下降,而钙化相关因子(Runx2、骨钙素)表达上升;③压应力能促进终板软骨细胞的细胞外基质降解,基质金属蛋白酶3、基质金属蛋白酶13表达量增加;④细胞内Wnt/β-catenin信号通路表达出现异常,其特异性抑制因子硬骨素伴随着β-catenin的异常积累而表达下降。结果说明:在压应力作用下,终板软骨细胞内硬骨素的表达下降,导致Wnt/β-catenin信号通路激活,促进软骨终板的钙化、退变与细胞外基质的降解,进而促进椎间盘退变。BACKGROUND:Many factors can cause disc degeneration,including aging,nutritional deficiency,and mechanical factors.The mechanical load is considered to be a very important factor,but its mechanism is still unclear.OBJECTIVE: To investigate the role of sclerostin (SOST) and Wnt/β-catenin signaling pathways in inducing degeneration of endplate cartilage. METHODS: Cartilage endplate cells were extracted from 4-week-old male Sprague-Dawley rats. Compressive stress was applied to endplate chondrocytes in vitro using a mechanical loading apparatus, and the cell viability was determined by the cell counting kit-8 assay at 1, 3, 5, and 7 days after compression. Western blot, reverse transcription quantitative PCR, and cellular immunofluorescence techniques were employed to examine intracellular cartilage markers (Aggrecan and type II collagen) as well as calcification-related factors (Runx2 and osteocalcin). The expression of extracellular matrix degradation enzyme and genes related to the signaling pathway (SOST and β-catenin) was also analyzed. RESULTS AND CONCLUSION: Under compressive stress, the cell activity of endplate chondrocytes increased with both the duration and intensity of stress. Furthermore, the expression levels of Aggrecan and type II collagen decreased in endplate cells under compressive stress, while those of calcification-related factors (Runx2 and osteocalcin) increased. Additionally, compressive stress promoted extracellular matrix degradation in endplate chondrocytes, leading to an increase in matrix metalloproteinase 3 and matrix metalloproteinase 13 expression. Abnormalities were observed in the Wnt/β-catenin signaling pathway within these cells under compressive stress, characterized by a decrease in specific inhibitory factor SOST expression accompanied by abnormal accumulation of β-catenin. To conclude, decreased SOST expression in endplate chondrocytes under compressive stress activates the Wnt/β-catenin signaling pathway, thereby promoting calcification, degeneration and extracellular m

关 键 词：软骨终板 软骨细胞 SOST WNT/Β-CATENIN 压应力 椎间盘退变 

分 类 号：R496[医药卫生—康复医学] R318[医药卫生—临床医学] R681