成纤维细胞生长因子2在椎间盘退行性变中的作用及机制  

Role and mechanism of fibroblast growth factor 2 in intervertebral disc degeneration

作　　者：李磊[1] 王一范 施强慧 钟华建 Li Lei;Wang Yifan;Shi Qianghui;Zhong Huajian(Department of Orthopaedics,Quzhou Hospital(Quzhou People’s Hospital),Wenzhou Medical University,Quzhou 324000,Zhejiang,China;Department of Health Management,Changzheng Hospital,Naval Medical University,Shanghai 200003,China;Department of Orthopaedics,63680th Hospital of Chinese PLA,Jiangyin 214400,Jiangsu,China;Department of Orthopaedics,Changzheng Hospital,Naval Medical University,Shanghai 200003,China)

机构地区：[1]温州医科大学附属衢州医院(衢州市人民医院)骨科,衢州324000 [2]海军军医大学长征医院健康管理科,上海200003 [3]中国人民解放军63680部队医院骨科,江阴214400 [4]海军军医大学长征医院骨科,上海200003

出　　处：《脊柱外科杂志》2023年第6期403-410,共8页Journal of Spinal Surgery

摘　　要：目的探讨成纤维细胞生长因子2(FGF2)在椎间盘退行性变(IDD)中的作用及调控机制。方法收集青年(6月龄)小鼠正常椎间盘及老年(24月龄)小鼠发生退行性变的椎间盘组织,通过免疫组织化学染色和蛋白质印迹法检测FGF2在IDD中的表达变化。培养原代大鼠髓核细胞,采用促炎因子白细胞介素-1β(IL-1β)诱导退行性变,并予以FGF2重组蛋白、FGF2中和抗体、FGF2干扰RNA等处理,通过流式细胞术、实时荧光定量PCR及蛋白质印迹法检测髓核细胞凋亡、细胞外基质(ECM)代谢相关基因表达及Wnt/β-catenin信号通路活性改变;采用Wnt/β-catenin信号通路抑制剂(PRI-724)处理FGF2重组蛋白刺激后的髓核细胞,明确Wnt/β-catenin信号通路对FGF2调控IDD的影响。结果FGF2表达水平在老年小鼠椎间盘组织及髓核细胞内均高于青年小鼠(P<0.05);FGF2重组蛋白促进正常髓核细胞凋亡及ECM分解相关基因表达,而FGF2中和抗体及干扰RNA在发生退行性变的细胞中作用相反。Wnt/β-catenin信号通路在FGF2重组蛋白或IL-1β刺激的髓核细胞内显著激活,PRI-724抑制该通路活性,可削弱FGF2对髓核细胞凋亡及ECM代谢的调控作用。结论发生IDD时,髓核细胞FGF2表达升高,通过Wnt/β-catenin信号通路促进髓核细胞凋亡及ECM降解,进一步加速IDD进程。Objective To investigate the role and the underlying mechanism of fibroblast growth factor 2(FGF2)in intervertebral disc degeneration(IDD).Methods The normal and degenerated intervertebral discs were collected from young(6 months old)and aged(24 months old)mice,respectively,and the expression of FGF2 during IDD was detected histologically by immunohistochemistry and Western blotting.Primary rat nucleus pulposus cells were isolated and cultured,then degeneration was induced using pro-inflammatory factor interleukin(IL)-1β,and treated with FGF2 recombinant protein,FGF2 neutralizing antibody,FGF2 interfering RNA,etc.The apoptosis,gene expression related to extracellular matrix(ECM)metabolism and Wnt/β-catenin signaling pathway activity were evaluated by flow cytometry,real-time PCR and Western blotting.Chemical inhibitor PRI-724 was used to inhibit the activity of Wnt/β-catenin signaling pathway in nucleus pulposus cells pretreated with FGF2,and the effect of Wnt/β-catenin signaling pathway on FGF2 modulating IDD was clarified.Results The expression level of FGF2 was significantly increased in degenerated intervertebral disc tissue and nucleus pulposus cells(P<0.05).Recombinant FGF2 protein promoted the expression of genes related to apoptosis and ECM degradation in normal nucleus pulposus cells,while inhibiting the expression of synthesis-related genes,FGF2 neutralizing antibodies and interfering RNAs showed opposite effects in degenerated nucleus pulposus cells.The Wnt/β-catenin signaling pathway was significantly activated in nucleus pulposus cells under FGF2 recombinant protein or IL-1βtreatment,what’s more,the effects of FGF2 on nucleus pulposus cells apoptosis and ECM metabolism were ameliorated with inhibition of Wnt/β-catenin signaling pathway through PRI-724.Conclusion The expression of FGF2 is elevated during IDD,which promotes apoptosis and ECM degradation of nucleus pulposus cells through the Wnt/β-catenin signaling pathway,which further accelerates the process of IDD.

关 键 词：椎间盘退行性变 成纤维细胞生长因子2 细胞凋亡 小鼠 

分 类 号：R342.2[医药卫生—基础医学]