胸椎黄韧带骨化患者的差异蛋白组学研究

Differential proteomics study of patients with sternal ossification of the ligamentum flavum

背景:针对胸椎黄韧带骨化发病原因及机制研究主要集中在功能基因组的研究,而从蛋白组学对其进行研究者甚少。目的:探讨胸椎黄韧带骨化患者骨化黄韧带与正常黄韧带组织差异蛋白质,寻找发病相关靶点蛋白。方法:收集3例胸椎黄韧带骨化患者的骨化黄韧带和相邻水平正常黄韧带组织,通过蛋白质提取、定量、同位素标记相对和绝对定量TMT技术标记肽段,运用液相色谱-串联质谱(LC-MS/MS)技术鉴定及筛选差异蛋白质。结果与结论:①共鉴定出差异表达蛋白质589种,其中表达上调蛋白质532种,表达下调蛋白质57种;②进一步分析鉴定出与炎性相关蛋白质共23种,其中蛋白CYBB、PODXL、S100A12、LXN、S100A10、GJA1、LIPA、FOLR2、CYBA、IFI16、ABCF1、HMGB1、RPS19、PRDX5、MAP2K3表达上调,蛋白KRT14、IL17D、IL1RAP、SLC7A2、RHOB、COMP、KRT6B表达下调;③这些炎性相关蛋白可能是参与胸椎黄韧带骨化疾病发生发展的靶点蛋白,尤其S100A10、FOLR2、S100A12、HMGB1这4种炎性相关蛋白在胸椎黄韧带骨化疾病发生发展中可能起重要作用,成为诊治的重要靶点。

BACKGROUND:The research on the pathogenesis and mechanism of thoracic ossification of the ligamentum flavum mainly focuses on the research of functional genome;however, less is reported on it from proteomics.OBJECTIVE: To investigate the differential proteins between ossified ligamentum flavum and normal ligamentum flavum in patients with thoracic ossification ofthe ligamentum flavum, and to search for disease-related target proteins.METHODS: Ossified ligamentum flavum and adjacent normal ligamentum flavum samples were collected from three patients with thoracic ossification of theligamentum flavum, and detected through protein extraction, quantification, isotope labeling for relative and absolute quantitative technique-labeled peptides.Differentially expressed proteins were identified and screened by liquid chromatography-tandem mass spectrometry.RESULTS AND CONCLUSION: Totally 589 kinds of differentially expressed proteins were identified, including 532 up-regulated proteins and 57 down-regulatedproteins. Further analysis identified 23 kinds of proteins related to inflammation, among which, the expression of CYBB, PODXL, S100A12, LXN, S100A10, GJA1,LIPA, FOLR2, CYBA, IFI16, ABCF1, HMGB1, RPS19, PRDX5, MAP2K3 was up-regulated, and the expression of KRT14, IL17D, IL1RAP, SLC7A2, RHOB, COMP, KRT6Bwas down-regulated. These inflammation-related proteins may be important diagnostic and therapeutic targets involved in the occurrence and developmentof thoracic ossification of the ligamentum flavum.