A transcriptomic study of probenecid on injured spinal cords in mice
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 th...
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doaj-c624dc50327d4e86be8d8817a3ff853d2020-11-25T02:37:12ZengPeerJ Inc.PeerJ2167-83592020-01-018e836710.7717/peerj.8367A transcriptomic study of probenecid on injured spinal cords in miceYu-Xin Zhang0Sai-Nan Wang1Jing Chen2Jian-Guo Hu3He-Zuo Lü4Clinical Laboratory, The First Affiliated Hospital of Bengbu Medical College, Bengbu, ChinaClinical Laboratory, The First Affiliated Hospital of Bengbu Medical College, Bengbu, ChinaClinical Laboratory, The First Affiliated Hospital of Bengbu Medical College, Bengbu, ChinaClinical Laboratory, The First Affiliated Hospital of Bengbu Medical College, Bengbu, ChinaClinical Laboratory, The First Affiliated Hospital of Bengbu Medical College, Bengbu, ChinaBackground 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).https://peerj.com/articles/8367.pdfProbenecidSpinal cord injuryRNA sequencingMice |
collection |
DOAJ |
language |
English |
format |
Article |
sources |
DOAJ |
author |
Yu-Xin Zhang Sai-Nan Wang Jing Chen Jian-Guo Hu He-Zuo Lü |
spellingShingle |
Yu-Xin Zhang Sai-Nan Wang Jing Chen Jian-Guo Hu He-Zuo Lü A transcriptomic study of probenecid on injured spinal cords in mice PeerJ Probenecid Spinal cord injury RNA sequencing Mice |
author_facet |
Yu-Xin Zhang Sai-Nan Wang Jing Chen Jian-Guo Hu He-Zuo Lü |
author_sort |
Yu-Xin Zhang |
title |
A transcriptomic study of probenecid on injured spinal cords in mice |
title_short |
A transcriptomic study of probenecid on injured spinal cords in mice |
title_full |
A transcriptomic study of probenecid on injured spinal cords in mice |
title_fullStr |
A transcriptomic study of probenecid on injured spinal cords in mice |
title_full_unstemmed |
A transcriptomic study of probenecid on injured spinal cords in mice |
title_sort |
transcriptomic study of probenecid on injured spinal cords in mice |
publisher |
PeerJ Inc. |
series |
PeerJ |
issn |
2167-8359 |
publishDate |
2020-01-01 |
description |
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). |
topic |
Probenecid Spinal cord injury RNA sequencing Mice |
url |
https://peerj.com/articles/8367.pdf |
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