Huang Zhixiang, Li Yinyu, Hu Guozhen, Ding Jiali, Liu Meng, Huang Yukai, Huang Xuechan, Li Tianwang
Department of Rheumatology and Immunology, The Affiliated Guangdong Second Provincial General Hospital of Jinan University, Guangzhou 510317, China; Guangdong Medical University, Zhanjiang 524023, China; The Second School of Clinical Medicine, Southern Medical University, Guangzhou 510515, China.
Department of Rheumatology and Immunology, The Affiliated Guangdong Second Provincial General Hospital of Jinan University, Guangzhou 510317, China.
Bone. 2025 Nov;200:117591. doi: 10.1016/j.bone.2025.117591. Epub 2025 Jul 14.
Excessive osteogenesis in bone marrow mesenchymal stem cells (BMSCs) contributes to the ectopic ossification associated with ankylosing spondylitis (AS), yet the underlying mechanisms are not fully understood. Integrin-linked kinase (ILK) plays an important role in the inflammatory process of AS, but its expression and effects on osteophytogenesis require further evaluation. Hence, we aimed to explore the role and mechanisms of ILK in the syndesmophyte formation of AS. After establishing the BMSC lines, the mineralization potential of BMSCs from AS patients (AS-BMSCs) was found to be greater than BMSCs of healthy volunteers (HV-BMSCs). The expression of ILK was consistent with the osteogenic hyperactivity of AS-BMSCs. Additionally, knockdown of ILK using small interfering ribonucleic acid suppressed osteogenic differentiation in BMSCs. Conversely, ILK upregulation via lentiviral transfection promoted their osteogenesis. The activity of protein kinase B (Akt)/ glycogen synthase kinase-3β (GSK-3β)/ β-catenin pathway in AS-BMSCs was higher than HV-BMSCs after osteogenic induction, while ILK overexpression further activated this axis. Besides, the osteogenic medium enhanced the nuclear translocation of β-catenin only in AS-BMSCs. Animal experiments revealed that the size and number of osteophytes progressively increased in a time-dependent manner in ankylosing enthesitis mice. Moreover, the expression of ILK in entheseal BMSCs was higher at week 24 and week 32 than at week 8, and this elevated expression positively correlated with osteophyte development. These findings indicate that increased ILK leads to excessive mineralization in AS-BMSCs via the activation of the Akt/GSK-3β/β-catenin pathway, resulting in ectopic ossification in AS patients.
骨髓间充质干细胞（BMSCs）中过度的骨生成导致与强直性脊柱炎（AS）相关的异位骨化，但其潜在机制尚未完全阐明。整合素连接激酶（ILK）在AS的炎症过程中起重要作用，但其表达及对骨赘形成的影响尚需进一步评估。因此，我们旨在探讨ILK在AS韧带骨赘形成中的作用及机制。建立BMSC系后，发现AS患者的BMSCs（AS-BMSCs）的矿化潜能大于健康志愿者的BMSCs（HV-BMSCs）。ILK的表达与AS-BMSCs的成骨活性一致。此外，使用小干扰核糖核酸敲低ILK可抑制BMSCs的成骨分化。相反，通过慢病毒转染上调ILK可促进其成骨。成骨诱导后，AS-BMSCs中蛋白激酶B（Akt）/糖原合酶激酶-3β（GSK-3β）/β-连环蛋白通路的活性高于HV-BMSCs，而ILK过表达进一步激活该信号轴。此外，成骨培养基仅在AS-BMSCs中增强了β-连环蛋白的核转位。动物实验显示，强直性附着点炎小鼠的骨赘大小和数量随时间呈渐进性增加。此外，附着点处BMSCs中ILK的表达在第24周和第32周高于第8周，且这种表达升高与骨赘形成呈正相关。这些发现表明，ILK增加通过激活Akt/GSK-3β/β-连环蛋白通路导致AS-BMSCs过度矿化，从而导致AS患者出现异位骨化。
