Wang Dan, Wang Limei, Han Jie, Zhang Zaili, Fang Bo, Chen Fengshou
Department of Anesthesiology, The First Hospital of China Medical University, Shenyang, China.
Front Genet. 2021 Apr 27;12:650180. doi: 10.3389/fgene.2021.650180. eCollection 2021.
Spinal cord ischemia/reperfusion injury (SCII) is a catastrophic complication involved with cardiovascular, spine, and thoracic surgeries and can lead to paraplegia. Nevertheless, the molecular mechanism of SCII remain ill-defined.
Expression profiling (GSE138966) data were obtained from GEO database. Then, differentially expressed (DE) lncRNAs and DEmRNAs were screened out with < 0.05, and | fold change| > 1.5. Aberrant miRNAs expression in SCII was obtained from PubMed. Functional enrichment analysis of overlapping DEmRNAs between predicted mRNAs in miRDB database and DEmRNAs obtained from GSE138966 was performed using cluster Profiler R package. The lncRNA-miRNA-mRNA competitive endogenous RNA (ceRNA) network was established in light of ceRNA theory. The key lncRNAs in the ceRNA network were identified by topological analysis. Subsequently, key lncRNAs related ceRNA-pathway network and transcription factors (TFs)-mRNAs network were constructed. Simultaneously, the expression levels of hub genes were measured via qRT-PCR.
The results in this study indicated that 76 miRNAs, 1373 lncRNAs, and 4813 mRNAs were differentially expressed in SCII. A SCII-related ceRNA network was constructed with 154 ncRNAs, 139 mRNAs, and 51 miRNAs. According topological analysis, six lncRNAs (NONRATT019236.2, NONRATT009530.2, NONRATT026999.2, TCONS_00032391, NONRATT023112.2, and NONRATT021956.2) were selected to establish the ceRNA-pathway network, and then two candidate hub lncRNAs (NONRATT009530.2 and NONRATT026999.2) were identified. Subsequently, two lncRNA-miRNA-mRNA regulatory axes were identified. NONRATT026999.2 and NONRATT009530.2 might involve SCII via miR-20b-5p/Map3k8 axis based on the complex ceRNA network. SP1 and Hnf4a acting as important TFs might regulate Map3k8. Furthermore, qRT-PCR results showed that the NONRATT009530.2, NONRATT026999.2, Map3k8, Hfn4a, and SP1 were significantly upregulated in SCII of rats, while the miR-20b-5p was downregulated.
Our results offer a new insight to understand the ceRNA regulation mechanism in SCII and identify highlighted lncRNA-miRNA-mRNA axes and two key TFs as potential targets for prevention and treatment of SCII.
脊髓缺血/再灌注损伤（SCII）是一种与心血管、脊柱和胸科手术相关的灾难性并发症，可导致截瘫。然而，SCII的分子机制仍不清楚。
从基因表达综合数据库（GEO数据库）获取表达谱（GSE138966）数据。然后，筛选出差异表达（DE）lncRNAs和DEmRNAs，设定条件为P值<0.05，且|倍数变化|>1.5。从PubMed获取SCII中异常的miRNAs表达情况。使用cluster Profiler R包对miRDB数据库中预测的mRNA与从GSE138966获得的DEmRNAs之间的重叠DEmRNAs进行功能富集分析。根据竞争性内源RNA（ceRNA）理论构建lncRNA-miRNA-mRNA ceRNA网络。通过拓扑分析确定ceRNA网络中的关键lncRNAs。随后，构建与ceRNA途径网络和转录因子（TFs）-mRNAs网络相关的关键lncRNAs。同时，通过qRT-PCR检测枢纽基因的表达水平。
本研究结果表明，在SCII中76个miRNAs、1373个lncRNAs和4813个mRNAs差异表达。构建了一个与SCII相关的ceRNA网络，包含15个lncRNAs、139个mRNAs和51个miRNAs。根据拓扑分析，选择6个lncRNAs（NONRATT019236.2、NONRATT009530.2、NONRATT026999.2、TCONS_00032391、NONRATT023112.2和NONRATT021956.2）建立ceRNA途径网络，然后鉴定出两个候选枢纽lncRNAs（NONRATT009530.2和NONRATT026999.2）。随后，鉴定出两个lncRNA-miRNA-mRNA调控轴。基于复杂的ceRNA网络，NONRATT026999.2和NONRATT009530.2可能通过miR-20b-5p/Map3k8轴参与SCII。作为重要TFs的SP1和Hnf4a可能调节Map3k8。此外，qRT-PCR结果显示，在大鼠SCII中，NONRATT009530.2、NONRATT026999.2、Map3k8、Hfn4a和SP1显著上调，而miR-20b-5p下调。
我们的结果为理解SCII中的ceRNA调控机制提供了新的见解，并鉴定出突出的lncRNA-miRNA-mRNA轴和两个关键TFs作为SCII预防和治疗的潜在靶点。
