发育性颈椎管狭窄人群血清差异蛋白质组学分析  

Serum differential proteomic analysis of developmental cervical canal stenosis

作　　者：卜献忠 钟远鸣[2] 卜保献 李记天 汪利合[1] 李慧英[1] 杨汉立 许伟 Bu Xianzhong;Zhong Yuanming;Bu Baoxian;Li Jitian;Wang Lihe;Li Huiying;Yang Hanli;Xu Wei(First Affiliated Hospital of Henan University of Chinese Medicine,Zhengzhou 450000,Henan Province,China;First Affiliated Hospital of Guangxi University of Chinese Medicine,Nanning 530023,Guangxi Zhuang Autonomous Region,China;Luoyang Orthopedic Hospital,Luoyang 471002,Henan Province,China)

机构地区：[1]河南中医药大学第一附属医院,河南省郑州市450000 [2]广西中医药大学第一附属医院,广西壮族自治区南宁市530023 [3]河南省洛阳正骨医院(河南省骨科医院),河南省洛阳市471002

出　　处：《中国组织工程研究》2024年第34期5432-5439,共8页Chinese Journal of Tissue Engineering Research

基　　金：国家自然科学基金项目(81760874,82260942),项目负责人:钟远鸣;广西中医药重点学科建设项目(GZXK-Z-20-21),项目负责人:钟远鸣;广西重点研发计划(桂科AB29159018),项目负责人:钟远鸣;河南省中医药科学研究专项课题(20-21ZY2083),项目负责人:卜保献;河南省中医药科学研究专项课题(2023ZXZX1003),项目认责人:卜献忠;洛阳市2022-2024年度中医重点学科-中西医结合医学(脊柱外科),项目负责人:卜保献。

摘　　要：背景:正常健康人与发育性颈椎管狭窄(气虚血瘀证)人群之间的血清特异性生物学标志物尚未完全明确。目的:寻找并鉴定气虚血瘀证发育性颈椎管狭窄的潜在生物学标志物。方法:分别采集发育性颈椎管狭窄(气虚血瘀证)人群血清9例、健康正常人群血清8例,采用同位素相对标记与绝对定量技术串联质谱标签联合液相色谱-串联质谱筛选2组人群血清的差异蛋白及鉴定,使用Western Blot法对肌球蛋白轻链3(MYL3)差异蛋白进行验证。结果与结论:(1)串联质谱标签技术:共鉴定出61个差异蛋白(P <0.05),其中与健康正常人群组比,补体成分C1q受体、载脂蛋白A4、C-C基序趋化因子配体18等14种差异蛋白明显上调,而肌球蛋白轻链3、线粒体翻译延伸因子、核仁磷酸蛋白1等47种差异蛋白明显下调;(2)基因富集分析:这些差异蛋白可能参与染色体组织、线粒体膜组织、肌肉系统过程的调节等分子功能;(3)蛋白相互作用网络分析:健康正常人群与发育性颈椎管狭窄(气虚血瘀证)人群之间的共同差异蛋白中补体C1q受体、载脂蛋白A4、C-C基序趋化因子配体18、肌球蛋白轻链3、线粒体翻译延伸因子、核仁磷酸蛋白1等38种位于功能网络节点,且与颈椎骨能量代谢、骨细胞产生、破骨细胞的形成等系统关系密切;(4)运用Western Blot法对主要差异蛋白肌球蛋白轻链3进行验证:其验证结果与蛋白组学结果相一致;(5)结论:通过绝对定量技术联合液相色谱-串联质谱技术发现健康正常人群与发育性颈椎管狭窄(气虚血瘀证)人群之间的血清差异表达蛋白,并初步推测肌球蛋白轻链3可能是发育性颈椎管狭窄(气虚血瘀证)人群的特异性血清标志物。BACKGROUND:Serum-specific biomarkers between normal healthy individuals and populations with developmental cervical canal stenosis(Qi deficiency and blood stasis syndrome)have not been fully defined.OBJECTIVE:To screen and identify the potential biomarkers of developmental cervical canal stenosis with Qi deficiency and blood stasis.METHODS:Serum samples were collected from nine patients with developmental cervical canal stenosis with Qi deficiency and blood stasis and eight healthy people.Differentially expressed proteins in serum were screened and identified using isotope relative labeling and absolute quantification combined with liquid chromatography tandem mass spectrometry.Western blot was used to verify some significant differentially expressed proteins.RESULTS AND CONCLUSION:A total of 61 differentially expressed proteins(P<0.05)were identified using tandem mass spectrometry techniques.Compared with the healthy normal population group,14 differentially expressed proteins such as complement component C1q receptor,apolipoprotein A4,and C-C motif chemokine ligand 18 were significantly upregulated,while 47 differentially expressed proteins such as myosin light chain 3,mitochondrial translation elongation factor,and nucleolar phosphoprotein 1 were significantly downregulated.The results of gene ontology enrichment analysis indicated that these differentially expressed proteins might participate in molecular functions such as regulation of chromosomal tissue,mitochondrial membrane tissue,and muscle system processes.Protein-protein interaction network analysis showed that 38 common differential proteins,including complement component C1q receptor,apolipoprotein A4,C-C motif chemokine ligand 18,myosin light chain 3,mitochondrial translation elongation factor,and nucleolar phosphoprotein 1,were located at functional network nodes between healthy normal individuals and those with developmental cervical canal stenosis(Qi deficiency and blood stasis syndrome),and were closely related to the local energy metabolism of

关 键 词：发育性颈椎管狭窄 气虚血瘀证 串联质谱标签 血清蛋白标志物 蛋白组学 

分 类 号：R496[医药卫生—康复医学] R318[医药卫生—临床医学] R681