腰椎间盘脱出症非手术治疗前后外周血基因表达变化特征及意义

戴国钢; 王一; 廖仕川; 夏姣; 王丰; 黄雷; 杜万里; 田国刚; 文江; 李涛

doi:10.12182/20210960303

腰椎间盘脱出症非手术治疗前后外周血基因表达变化特征及意义

Characteristics and Significance of Gene Expression Changes in Peripheral Blood of Lumbar Disc Extrusion Patients before and after Nonoperative Treatment

摘要

摘要:
目的研究腰椎间盘脱出患者外周血基因表达特征及非手术治疗对其表达的影响。
方法采用基因芯片半定量测定初步筛选腰椎间盘脱出患者和健康对照者外周血中的差异表达基因，以及患者在非手术治疗后这些差异表达基因的变化趋势，通过富集分析研究差异表达基因的功能特征，通过网络分析找出基因异常表达的关键基因，采用qRT-PCR定量检测验证这些基因在患者和健康对照样本中的表达情况以及非手术治疗对这些差异表达基因的影响。
结果在腰椎间盘脱出患者和健康对照组的外周血中发现153个差异表达基因，其中131个基因表达上调，22个基因表达下调；富集分析显示大部分差异表达基因与免疫以及炎症反应相关；网络分析显示Toll样受体4（toll-like receptor 4, TLR4）、基质金属肽酶9（matrix metallopeptidase 9, MMP9）、髓过氧化物酶（myeloperoxidase, MPO)、抗菌肽（cathelicidin antimicrobial peptide, CAMP）、resistin基因（RETN）和Toll样受体5（toll-like receptor 5, TLR5）是蛋白互作网络中的关键基因，这些关键基因均被富集到了免疫、炎症反应相关的条目。非手术治疗后，患者疼痛减轻，在这153个差异表达基因中，TLR5、白介素1受体拮抗剂（interleukin 1 receptor antagonist, IL1RN）和溶质载体家族8成员A1（solute carrier family 8 member A1, SLC8A1）在治疗后表达下调。qRT-PCR结果显示：患者外周血中TLR4、MMP9、MPO、CAMP、RETN、TLR5、IL1RN和SLC8A1表达水平高于健康对照组（P<0.05）；治疗后与治疗前比较，TLR5、IL1RN和SLC8A1表达水平降低（P<0.05）。
结论腰椎间盘脱出患者外周血基因表达特征主要是免疫和炎症反应相关基因表达失调，其中TLR4、MMP9、MPO、CAMP、RETN和TLR5这些与免疫和炎症反应相关的基因在腰椎间盘脱出患者外周血基因表达失调中起关键作用，非手术治疗疗效的获得可能与患者外周血中过度表达的TLR5、IL1RN和SLC8A1下调相关。

Abstract:
Objective To define the gene expression characteristics in the peripheral blood of patients with lumbar disc extrusion (LDE) and the effect of nonoperative treatment on the gene expression.
Methods DNA microarray was used to identify semi-quantitatively the differentially expressed genes (DEGs) in the peripheral blood of patients with LDE and that of the healthy controls and the variation trend of these DEGs after nonoperative treatment. Enrichment analysis was done to reveal the functional characteristics of these DEGs, and network analysis was done to identify key genes that contribute to gene dysregulation. The levels of these key genes were measured by qRT-PCR to examine their expression in LDE patients and the controls, and the effect of nonoperative treatment on the expression level.
Results We identified 153 DEGs in the peripheral blood of LDE patients and healthy controls, including 131 upregulated genes and 22 downregulated genes. Enrichment analysis revealed that most of the DEGs were related to immunity and the inflammatory response. Network analysis revealed that toll-like receptor 4 (TLR4), matrix metallopeptidase 9 (MMP9) and myeloperoxidase (MPO), cathelicidin antimicrobial peptide (CAMP), resistin (RETN), toll-like receptor 5 (TLR5) were the key genes in the protein-protein interaction network. These key genes were all enriched into the terms releated to immunity and the inflammatory response. The patients experienced pain relief after nonoperative treatment. Among the 153 DEGs, TLR5 , interleukin 1 receptor antagonist (IL1RN) and solute carrier family 8 member A1 (SLC8A1) were downregulated after nonoperative treatment. qRT-PCR revealed that the levels of TLR4, MMP9, MPO, CAMP, RETN, TLR5, IL1RN and SLC8A1 in the peripheral blood of the LDE patients were higher than those of the healthy control group (P<0.05). In addition, TLR5, IL1RN and SLC8A1 expression levels decreased after treatmentin in comparison with the levels before treatment (P<0.05).
Conclusion Gene expression in the peripheral blood of LDE patients was characterized by the dysregulation of immune and inflammatory response-related genes, among which, TLR4, MMP9, MPO, CAMP, RETN and TLR5, the genes relevant to immune and inflammatory response, played a key role in the dysregulation of gene expression in the peripheral blood of LDE patients. The outcome of non-operative treatment may be related to the downregulation of the overexpressed TLR5, IL1RN and SLC8A1 in the peripheral blood of patients.

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