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血清蛋白质组的比较分析:鉴定与腰椎间盘突出症所致坐骨神经痛相关的蛋白质。

Comparative analysis of serum proteomes: Identification of proteins associated with sciatica due to lumbar intervertebral disc herniation.

作者信息

Xie Peigen, Liu Bin, Chen Ruiqiang, Yang Bu, Dong Jianwen, Rong Limin

机构信息

Department of Spine Surgery, The Third Affiliated Hospital of Sun Yat-sen University, Guangzhou, Guangdong 510630, P.R. China.

出版信息

Biomed Rep. 2014 Sep;2(5):693-698. doi: 10.3892/br.2014.295. Epub 2014 Jun 16.

Abstract

Lumbar intervertebral disc herniation (LDH) is one of the most common orthopedic conditions that can cause lower back pain and sciatica. However, the pathogenesis of LDH is poorly understood. The aim of the present study was to use proteomic analysis of blood samples to establish whether there are serum proteins associated with LDH, which may be useful in elucidating LDH pathogenesis. The ultimate aim was to develop a simple technique for the diagnosis of LDH based on the blood samples of patients with sciatica. The study used comparative analysis of serum proteomes associated with sciatica due to LDH. A total of 30 LDH patients with sciatica, receiving treatment between August and December 2007, were selected as the experimental group (or LDH group). A total of 2 ml of blood was obtained from each of the 30 patients in the LDH group and from 30 healthy volunteers, who constituted the control group. Two-dimensional electrophoresis of the blood samples was conducted, distinct protein spots were identified by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry and proteins associated with LDH were detected. An enzyme-linked immunosorbent assay (ELISA) was developed to screen for the LDH proteins and was tested on the sera of a second test and control group that included 10 patients with LDH and 10 healthy subjects, respectively. Based on signal intensity, the expression levels of 6 proteins on the dielectrophoretogram were found to be significantly associated with LDH. The identities of the LDH proteins were upregulated apolipoprotein-L1 (APO-L1) and two types of serum albumin precursors, and downregulated apolipoprotein M (APO-M), tetranectin (TN) and immunoglobulin light chain (IGL). Further ELISA experiments confirmed that there were increased serum levels of 4 out of the 6 proteins in patients with sciatica due to LDH, which was statistically different compared to the healthy subjects. In conclusion, these results suggest that serum APO-L1, TN, APO-M and IGL may serve as LDH biomarkers.

摘要

腰椎间盘突出症(LDH)是最常见的可导致下背部疼痛和坐骨神经痛的骨科疾病之一。然而,LDH的发病机制尚不清楚。本研究的目的是通过对血液样本进行蛋白质组学分析,确定是否存在与LDH相关的血清蛋白,这可能有助于阐明LDH的发病机制。最终目的是基于坐骨神经痛患者的血液样本开发一种简单的LDH诊断技术。该研究对与LDH所致坐骨神经痛相关的血清蛋白质组进行了比较分析。选取2007年8月至12月期间接受治疗的30例伴有坐骨神经痛的LDH患者作为实验组(或LDH组)。从LDH组的30例患者和30名健康志愿者(构成对照组)中分别采集2ml血液。对血液样本进行二维电泳,通过基质辅助激光解吸/电离飞行时间质谱鉴定不同的蛋白质斑点,并检测与LDH相关的蛋白质。开发了一种酶联免疫吸附测定(ELISA)来筛选LDH蛋白,并在第二个测试组和对照组的血清上进行测试,该组分别包括10例LDH患者和10名健康受试者。基于信号强度,发现6种蛋白质在介电电泳图上的表达水平与LDH显著相关。LDH蛋白的鉴定结果为上调的载脂蛋白-L1(APO-L1)和两种血清白蛋白前体,以及下调的载脂蛋白M(APO-M)、四连蛋白(TN)和免疫球蛋白轻链(IGL)。进一步的ELISA实验证实,LDH所致坐骨神经痛患者血清中6种蛋白质中有4种水平升高,与健康受试者相比有统计学差异。总之,这些结果表明血清APO-L1、TN、APO-M和IGL可能作为LDH的生物标志物。

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