腰椎间盘突出症的血清蛋白质组学研究

Comparative Analysis of Serum Proteomes of Lumbar Intervertebral Disc Herniation

【目的】找出有坐骨神经痛的腰椎间盘突出症患者血清蛋白质组最明显的改变,揭示椎间盘突出可能的病理机制。【方法】选择有坐骨神经痛的腰椎间盘突出症患者10例作为病例组,志愿者10例作为对照组,取血,提取分离纯化血清蛋白,双向电泳,找到表达上差异最明显蛋白质,进行质谱鉴定。所得差异蛋白应用酶联免疫吸附试验对另外10例腰椎间盘突出症患者及10例正常人血清表达浓度进行检测。【结果】在双向电泳图谱上有6个蛋白质的表达在两组间存在明显差异。其中载脂蛋白L1以及两个白蛋白前体表达升高,载脂蛋白M、四连接素、免疫球蛋白轻链表达降低,酶联免疫吸附试验证实6种蛋白中的4种在血清里高表达,并与正常人存在差异。【结论】实验结果提示载脂蛋白L1、载脂蛋白M、四连接素、免疫球蛋白轻链的血清学水平可能作为腰椎间盘突出症的生物标志物,并可能进一步揭示腰椎间盘突出症的发病机制。

[Objective] To use proteomic analysis of blood samples to establish if there are serum proteins associated with lumbar intervertebral disc herniation （LDH）, which in addition, would elucidate the pathogenesis of LDH. [Methods] Ten LDH patients with sciatica were selected as the experimental group. Two ml of blood was drawn from each of the 10 patients in the experimental group as well as from 10 normal volunteers which constituted the control group. Two-dimensional eleetrophoresis （2- DE） of the blood samples was conducted, distinct protein spots were identified by matrix-assisted laser desorption/ionization time- of-flight mass spectrometry （MALDI-TOF MS） and six proteins associated with intervertebral disc herniation were detected. An enzyme-linked-immunosorbent assay （ELISA） was developed to screen for these LDH proteins, and was tested on the serum of a second test group of ten patients with LDH, and a second control group of 10 normal subjects. [Results] The expression of 6 proteins on the dielectrophoretogram was found to be of significant association to the patients with LDH. The identities of these LDH up-regulated proteins were： up-regulated apolipoprotein-L1 and two types of serum albumin precursor, and down-regulated apolipoprotein M, tetranectin, and immunoglobulin light chain （IGL）. Further ELISA confirmed that there were increased serum levels of 4 out of the 6 proteins in the patients with sciatica due to LDH, which were of statistical difference compared with those of normal subjects. [ Conclusion ] These results imply that serum APO-L1, tetranectin, APO-M, and IGL maybe serve as biomarkers for LDH, which would elucidate the pathogenesis of LDH.