胱抑蛋白C在腰椎间盘突出症患者脑脊液中的改变 可否作为神经源性疼痛的标志物?

Alteration of cystatin C in cerebrospinal fluid of patients with lumbar intervertebrai disc herniation:Can it be a marker for neuropathic pain?

目的:对有坐骨神经痛的腰椎间盘患者脑脊液进行蛋白质组学分析,找出改变最明显的蛋白质,揭示其与神经源性疼痛的关系。方法:选择2004-05／09在上海交通大学附属第六人民医院骨科住院的有坐骨神经痛的突出型腰椎间盘突出症患者10例作为腰椎间盘突出症组,每例取2 mL脑脊液,与对照组10例志愿者的脑脊液标本经丙酮沉淀后,依次进行等电聚焦和十二烷基磺酸钠-聚丙烯酰胺凝胶电泳; 将所得凝胶银染后扫描,用PDQuest图像分析软件(Bio-Rad)进行处理, 找到表达上差异最明显蛋白质,进行质谱鉴定。结果:20例均进入结果分析。①在双向电泳图谱上有15个蛋白质的表达在两组间存在明显差异。其中9个蛋白质在腰椎间盘突出症组表达升高,6个降低。②用液相色谱-电喷雾离子阱-质谱鉴定其中改变最明显的蛋白质为胱抑蛋白C。③酶联免疫吸附试验证实在腰椎间盘突出引起坐骨神经痛患者的脑脊液中,胱抑蛋白C的浓度明显高于对照组 [(2．93±0．49),(2．26±0．34)mg／L,P<0．01]．结论:胱抑蛋白C在有坐骨神经痛的腰椎间盘突出症患者的脑脊液中的浓度明显升高,可以作为反映神经源性疼痛的一种标记物。

AIM： Proteomic analysis is carried out in the cerebrospinal tluid （CSF） in sciatica patients induced by lumbar intervertebral disk herniation （LIDH） to find the protein that presents with the most obvious change and to investigate its relation with neuropathic pain. METHODS：Ten lumbar disc protrusion patients with sciatica,who were treated at the Department of Orthopaedics, Shanghai Sixth People＇s Hospital, Jiaotong University from May to September 2004,were selected as LIDH grnup.Two mL CSF were gained from 10 patients and 10 normal volunteers in the control group were treated with isoelectric focusing （IEF） and sodium dedeeylsulphate polyacrylamide gel electrnphoresis（SDS-PAGE） after removing salt with acetone.Then the harvested gel images were scanned＇after sliver staining, and analyzed with PDQuest 2D-gel seftware （Bio-Rad） to find the obviously alternated proteins that used for mass spectrometry evaluation. RESULTS：Totally 20 patients were involved in the result analysis.① Fifteen expressions of protein on dielectrnphoretogram had significant difference between two groups.Expression of nine proteins in the LIDH group increased, and 6 decreased. ② The most significantly differential protein was identified as eystatin C by LC-IT-MS. ③Additional enzymelinked inununoserbent assay confirmed that the level, of cystatin C in CSF of the patients with sciatica due to lumbar intervertebral prolapse was significantly higher than that of control group [（2.93±0.49）, （2.26±0.34） mg/L, P 〈 0.01]. CONCLUSION：In the CSF of patients with sciatica due to lumbar intervertebral prolapse, the volume of cystatin C increases significantly, which can be used as a marker for neuropathic pain.