A transcriptomic study of probenecid on injured spinal cords in mice
A transcriptomic study of probenecid on injured spinal cords in mice
复制标题
丙磺舒对小鼠脊髓损伤的转录组学研究
DOI：
10.7717/peerj.8367
复制
发表时间：
2020-01-03
期刊：
影响因子：
2.7
通讯作者：
Lu, He-Zuo
中科院分区：
文献类型：
--
作者：
Zhang, Yu-Xin;Wang, Sai-Nan;Lu, He-Zuo
背景：最近的研究发现，概率对中枢神经系统损伤具有神经保护作用和修复作用。然而，它对急性脊髓损伤（SCI）中全基因组转录的影响仍然未知。在本研究中，RNA测序（RNA-SEQ）用于分析脊柱损伤后8小时基因转录的局部表达的影响。方法：无限的地平线撞击子在小鼠中执行偶发性SCI。 SCI模型是通过使用50 kdynes的力的杆（直径1.3毫米）制成的。假手术小鼠仅接受椎板切除术而没有造成损伤。将受伤的小鼠随机分配到对照（SCI_C）或ProbEnecid注入（SCI_P）组中。在后一组中，损伤后立即对probenecid药物进行腹膜内注射（0.5 mg/kg）。损伤或椎板切除术后八小时，两组的小鼠从小鼠中取出脊髓。提取总RNA并纯化以进行库制备和转录组测序。使用DESEQ软件分析了三组SHAM，SCI_C和SCI_P的差分基因表达式（DEG）。基因本体（GO）和基因和基因组的京都百科全书（KEGG）富集分析DEG是使用Goseq R包和Kobas软件进行的。使用实时定量逆转录酶聚合酶链反应验证RNA-seq结果。results：RNA-Seq表明，与SCI_C组相比，SCI_P组的DEG数量为641（286个上置和355个下调） 。根据GO分析，DEG最富含细胞外基质（ECM），胶原蛋白三聚体，蛋白质边界和序列特异性DNA结合。 KEGG分析表明，最丰富的途径包括：细胞粘附分子，白细胞跨内皮迁移，ECM受体相互作用，PI3K-AKT信号通路，造血细胞谱系，局灶性粘附，RAP 1信号通路等。进入序列读取存档（https://www.ncbi.nlm.nih.gov/sra/prjna554464）。
Background Recent studies have found that probenecid has neuroprotective and reparative effects on central nervous system injuries. However, its effect on genome-wide transcription in acute spinal cord injury (SCI) remains unknown. In the present study, RNA sequencing (RNA-Seq) is used to analyze the effect of probenecid on the local expression of gene transcription 8 h after spinal injury. Methods An Infinite Horizon impactor was used to perform contusive SCI in mice. The SCI model was made by using a rod (1.3 mm diameter) with a force of 50 Kdynes. Sham-operated mice only received a laminectomy without contusive injury. The injured mice were randomly assigned into either the control (SCI_C) or probenecid injection (SCI_P) group. In the latter group, the probenecid drug was intraperitoneally injected (0.5 mg/kg) immediately following injury. Eight hours after the injury or laminectomy, the spinal cords were removed from the mice in both groups. The total RNAs were extracted and purified for library preparation and transcriptome sequencing. Differential gene expressions (DEGs) of the three groups—sham, SCI_C and SCI_P—were analyzed using a DESeq software. Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analysis of DEGs were performed using a GOseq R package and KOBAS software. Real-time quantitative reverse-transcriptase polymerase chain reaction was used to validate RNA-Seq results. Results RNA-Seq showed that, compared to the SCI_C group, the number of DEGs was 641 in the SCI_P group (286 upregulated and 355 downregulated). According to GO analysis, DEGs were most enriched in extracellular matrix (ECM), collagen trimer, protein bounding and sequence specific DNA binding. KEGG analysis showed that the most enriched pathways included: cell adhesion molecules, Leukocyte transendothelial migration, ECM-receptor interactions, PI3K-Akt signaling pathways, hematopoietic cell lineages, focal adhesions, the Rap1 signaling pathway, etc. The sequence data have been deposited into the Sequence Read Archive (https://www.ncbi.nlm.nih.gov/sra/PRJNA554464).