何斌;沈皆亮;许圣茜;郝杰;胡侦明;

• 1:贵州省遵义市第一人民医院骨科二病区
• 2:重庆医科大学附属第一医院骨科

摘要(Abstract)：

[目的]探讨SDF-1/CXCR4信号轴对于退变椎间盘胞外基质的影响及其潜在的作用机制。[方法]分别从年龄相匹配的椎间盘突出症和脊柱骨折手术患者中获取椎间盘组织,通过术前磁共振(MRI)采用Pfirrmann分级对椎间盘样本进行退变程度分组。免疫组化和western blot检测两个组椎间盘组织SDF-1、CXCR4、MMP-3,MMP9,Collagen Ⅱ和Aggrecan的表达。对取自椎间盘突出症患者的组织进行分离,细胞原代培养。取第3~6代细胞加入100ng/ml SDF-1或转染CXCR4小干扰RNA(siRNA)后共培养24 h,荧光定量PCR和western blot检测CXCR4、Collagen Ⅱ和Aggrecan的表达,Annexin V/PI双染法检测细胞凋亡率。为进一步探讨其潜在的分子机制,NF-κB抑制剂PDTC(20μmol/L)或CXCR4-siRNA转染髓核细胞后,观察NF-κB的主要基团P65磷酸化水平(p-P65)及核转移情况,胞外基质的表达和细胞凋亡水平是否发生改变。[结果]通过MRI的Pfirrmann分级标准,椎间盘突出组的分级为Grade Ⅲ~Ⅴ级,而脊柱骨折组的分级为Grade Ⅰ、Ⅱ级,分别定义为退变组和正常组。通过检测两组未进行处理的椎间盘组织发现,SDF-1、CXCR4、MMP-3和MMP-9在退变组中表达增高,而胞外基质的主要成分Collagen Ⅱ和Aggrecan表达则在退变组织中明显降低。而体外细胞实验结果显示,SDF-1处理后CXCR4的表达明显增高,同时伴随细胞凋亡的增高,而Collagen Ⅱ、Aggrecan的表达则显著降低,但这一作用可以随着CXCR4被siRNA靶向沉默后受到明显抑制。作者进一步采用PDTC或CXCR4-siRNA处理细胞后发现,SDF-1可以明显提高p-P65表达水平,促进P65基团的核转移,而这些作用随着CXCR4表达降低而受到抑制。此外PDTC抑制NF-κB活性后,细胞凋亡水平明显下降,而Collagen Ⅱ和Aggrecan的表达则明显上升。[结论]SDF-1/CXCR4信号轴在退变椎间盘中表达增高,它可以促进胞外基质的降解,增加细胞的凋亡水平,其潜在的机制通过调控NF-κB的活性得以实现。SDF-1/CXCR4信号轴有望成为治疗椎间盘退变疾病的潜在靶点。

关键词(KeyWords)： 椎间盘退变;SDF-1/CXCR4;NF-κB;胞外基质;细胞凋亡

Abstract:

Keywords:

基金项目(Foundation): 国家自然科学基金面上项目(编号:81171751,81372003)

作者(Author): 何斌;沈皆亮;许圣茜;郝杰;胡侦明;

Email:

DOI:

参考文献(References)：

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