急性期神经根型颈椎病患者的血清差异蛋白组学分析

Analysis of serum differential proteomics in patients with acute cervical spondylotic radiculopathy

背景:正常健康人向急性期神经根型颈椎病转化的具体分子机制尚未完全明确,需进一步深入研究。目的:探讨正常健康人群与急性期神经根型颈椎病患者的血清蛋白组学差异表达,并寻找及鉴定两者之间的潜在特异性血清标志物。方法:收集急性期神经根型颈椎病患者、正常健康人群血清各8例,应用绝对定量技术联合液相色谱-串联质谱技术进行蛋白组学筛选与分析,以期探寻与鉴定急性期神经根型颈椎病患者差异表达的血清蛋白质。结果与结论:①绝对定量技术筛查出有意义差异蛋白183种,鉴定出11种显著性差异蛋白质(P<0.05),其中与正常健康人组相比,急性期神经根型颈椎病组中人类白细胞抗原A、分泌珠蛋白家族1a成员1、蛋白4-羟基苯丙酮酸双加氧酶等3种差异蛋白显著上调,免疫球蛋白IgG3恒定区、皮肤因子、肌球蛋白轻链3等7种差异蛋白显著下调;②基因本体富集分析结果表明,这些差异蛋白参与了抗原结合、免疫球蛋白受体结合等分子功能;③蛋白相互作用网络分析表明,正常健康人群与急性期神经根型颈椎病之间的共同差异蛋白中HLA-A、HPD、PSMA3、DMKN、SCGB1A1、MYL3等6种位于功能网络节点,且与机体免疫、细胞炎症反应、能量代谢、机械压力等系统关系密切;④采用免疫印迹实验对显著性差异蛋白HLA-A、HPD、MYL3进行验证,其验证结果与蛋白组学结果相一致;⑤提示通过绝对定量技术联合液相色谱-串联质谱技术发现正常健康人群与急性期神经根型颈椎病患者之间的血清差异表达蛋白质,并初步认为HLA-A、HPD、MYL3可能是急性期神经根型颈椎病的特异性血清标志物,为进一步研究其发病机制提供了新方向。

BACKGROUND:The specific molecular mechanism of the transformation from normal healthy people to acute cervical spondylotic radiculopathy has not been clear,which needs to be further studied.OBJECTIVE:To investigate the differential expression of serum proteomics between normal healthy people and patients with acute cervical spondylotic radiculopathy,and to find and identify potential specific serum markers between them.METHODS:The serum samples of eight patients with acute cervical spondylotic radiculopathy and eight normal healthy people were collected,and the proteomic screening and analysis were performed by tandem mass tag combined with liquid chromatography-tandem mass spectrometry technology,in order to explore and identify serum proteins differentially expressed in patients with acute cervical spondylotic radiculopathy.RESULTS AND CONCLUSION:A total of 183 significantly differential proteins were screened by tandem mass tag technology,and 11 significantly differential proteins were identified(P<0.05).Compared with normal healthy people,three differential proteins were significantly up-regulated,including human leukocyte antigen-A,secretoglobin family 1a member 1,and protein 4-hydroxyphenylpyruvate dioxygenase,and seven differential proteins were significantly down-regulated,such as immunoglobulin heavy constant gamma 3,skin factor,and myosin light chain 3,in patients with acute cervical spondylotic radiculopathy.Gene ontology enrichment analysis showed that these differential proteins participated in antigen binding,immunoglobulin receptor binding and other molecular functions.Protein-protein interaction analysis showed that among the common differential proteins between normal healthy people and patients with acute cervical spondylotic radiculopathy,HLA-A,HPD,PSMA3,DMKN,SCGB1A1,and MYL3 were located at the nodes of the functional network,and were closely related to the systems of body immunity,cellular inflammatory response,energy metabolism,and mechanical pressure.The significantly differential proteins HLA-A,HPD and MYL3 were verified by western blot,and the results were consistent with those of proteomics.To conclude,tandem mass tag combined with liquid chromatography-tandem mass spectrometry technology can be used to find the differentially expressed proteins in serum between normal healthy people and patients with acute cervical spondylotic radiculopathy.It is preliminarily believed that HLA-A,HPD and MYL3 may be specific serum markers of acute cervical spondylotic radiculopathy,providing a new direction for further research on its pathogenesis.