ABSTRACT
We have previously reported that morroniside promoted motor activity after spinal cord injury (SCI) in rats. However, the mechanism by which morroniside induces recovery of injured spinal cord (SC) remains unknown. In the current study, RNA sequencing (RNA-seq) was employed to evaluate changes of gene expressions at the transcriptional level of the injured spinal cords in morroniside-administrated rats. Principal component analysis, analysis of enriched Gene Ontology (GO), enrichment analyses Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway, and other bioinformatics analyses were executed to distinguish differentially expressed genes (DEGs). The results of RNA-seq confirmed the anti-inflammatory and anti-apoptotic effects of morroniside on injured SC tissues, and provided the basis for additional research of the mechanisms involving the protective effects of morroniside on SCI.
Subject(s)
Anti-Inflammatory Agents/administration & dosage , Gene Expression Profiling/methods , Gene Regulatory Networks/drug effects , Glycosides/administration & dosage , Spinal Cord Injuries/drug therapy , Animals , Anti-Inflammatory Agents/pharmacology , Disease Models, Animal , Female , Gene Expression Regulation/drug effects , Gene Ontology , Glycosides/pharmacology , Principal Component Analysis , Random Allocation , Rats , Sequence Analysis, RNA , Spinal Cord Injuries/etiology , Spinal Cord Injuries/geneticsABSTRACT
Spinal cord injury (SCI) is a disabling condition that often leads to permanent neurological deficits without an effective treatment. Reactive oxygen species (ROS) produced during oxidative stress play a vital role in the pathogenesis following SCI. The antioxidant morroniside is the main active component of the Chinese medicine Cornus officinalis. In recent years, it has been reported that morroniside has therapeutic effects on damage to multiple organs mediated by oxidative damage, but the effect of morroniside on SCI has not been reported. The purpose of this study was therefore to assess the therapeutic effect of morroniside on SCI, and to identify its underlying mechanism by direct intragastric administration immediately after SCI. Our study showed that morroniside treatment improved the functional recovery of rats following SCI. This behavioral improvement was associated with the higher survival in neurons and oligodendrocytes following SCI, which increased the capacity of injured spinal cord (SC) to form myelin and repair tissue, eventually contributing to improved neurological outcome. Furthermore, our study found that oxygen free radicals increased and antioxidant enzyme activity decreased in the injured SC. Interestingly, morroniside treatment decreased oxygen free radical levels and increased antioxidant enzyme activities. Together, our results suggested that morroniside may be an effective treatment for improving outcomes following SCI, and that its antioxidant activity may be one of the mechanisms by which morroniside exerts neuroprotective effects on SCI.
Subject(s)
Glycosides/pharmacology , Neuroprotective Agents/pharmacology , Spinal Cord Injuries/drug therapy , Animals , Antioxidants/pharmacology , Cell Survival/drug effects , Cornus/chemistry , Female , Locomotion , Neurons/pathology , Oligodendroglia/pathology , Rats , Rats, Sprague-Dawley , Reactive Oxygen Species , Recovery of Function , Spinal Cord Injuries/pathologyABSTRACT
MicroRNAs (miRNAs) are involved in a series of pathology of spinal cord injury (SCI). Although, locally expressed miRNAs have advantages in studying the pathological mechanism, they cannot be used as biomarkers. The "free circulation" miRNAs can be used as biomarkers, but they have low concentration and poor stability in body fluids. Exosomal miRNAs in body fluids have many advantages comparing with free miRNAs. Therefore, we hypothesized that the specific miRNAs in the central nervous system might be transported to the peripheral circulation and concentrated in exosomes after injury. Using next-generation sequencing, miRNA profiles in serum exosomes of sham and subactue SCI rats were analyzed. The results showed that SCI can lead to changes of serum exosomal miRNAs. These changed miRNAs and their associated signaling pathways may explain the pathological mechanism of suacute SCI. More importantly, we found some valuable serum exosomal miRNAs for diagnosis and prognosis of SCI.
Subject(s)
Exosomes/genetics , MicroRNAs/metabolism , Spinal Cord Injuries/genetics , Animals , Gene Expression Profiling , RNA, Small Untranslated/metabolism , Rats , Real-Time Polymerase Chain Reaction , Spinal Cord Injuries/blood , Spinal Cord Injuries/metabolism , Spinal Cord Injuries/pathologyABSTRACT
Previous studies have shown that caspase-1 plays an important role in the acute inflammatory response of spinal cord injury (SCI). VX765, a novel and irreversible caspase1 inhibitor, has been reported to effectively intervene in inflammation. However, the effect of VX765 on genomewide transcription in acutely injured spinal cords remains unknown. Therefore, in the present study, RNAsequencing (RNASeq) was used to analyze the effect of VX765 on the local expression of gene transcription 8 h following injury. The differentially expressed genes (DEGs) underwent enrichment analysis of functions and pathways by Gene Ontology and Kyoto Encyclopedia of Genes and Genomes analyses, respectively. Parallel analysis of western blot confirmed that VX765 can effectively inhibit the expression and activation of caspase1. RNASeq showed that VX765 treatment resulted in 1,137 upregulated and 1,762 downregulated DEGs. These downregulated DEGs and their associated signaling pathways, such as focal adhesion, cytokinecytokine receptor interaction, leukocyte transendothelial migration, extracellular matrixreceptor interaction, phosphatidylinositol 3kinaseprotein kinase B, Rap1 and hypoxia inducible factor1 signaling pathway, are mainly associated with inflammatory response, local hypoxia, macrophage differentiation, adhesion migration and apoptosis of local cells. This suggests that the application of VX765 in the acute phase can improve the local microenvironment of SCI by inhibiting caspase1. However, whether VX765 can be used as a therapeutic drug for SCI requires further exploration. The sequence data have been deposited into the Sequence Read Archive (https://www.ncbi.nlm.nih.gov/sra/PRJNA548970).
Subject(s)
Anti-Inflammatory Agents, Non-Steroidal/pharmacology , Caspase Inhibitors/pharmacology , Dipeptides/pharmacology , Spinal Cord Injuries/drug therapy , Transcriptome/drug effects , para-Aminobenzoates/pharmacology , Animals , Anti-Inflammatory Agents, Non-Steroidal/therapeutic use , Caspase 1/metabolism , Caspase Inhibitors/therapeutic use , Dipeptides/therapeutic use , Female , Gene Expression Profiling , Mice , Mice, Inbred C57BL , Spinal Cord Injuries/genetics , Spinal Cord Injuries/metabolism , para-Aminobenzoates/therapeutic useABSTRACT
MicroRNAs (miRNAs) are involved in a series of pathology of spinal cord injury (SCI). Although, locally expressed miRNAs have advantages in studying the pathological mechanism, they cannot be used as biomarkers. The "free circulation" miRNAs can be used as biomarkers, but they have low concentration and poor stability in body fluids. Exosomal miRNAs in body fluids have many advantages comparing with free miRNAs. Therefore, we hypothesized that the specific miRNAs in the central nervous system might be transported to the peripheral circulation and concentrated in exosomes after injury. Using next-generation sequencing, miRNA profiles in serum exosomes of sham and subactue SCI rats were analyzed. The results showed that SCI can lead to changes of serum exosomal miRNAs. These changed miRNAs and their associated signaling pathways may explain the pathological mechanism of suacute SCI. More importantly, we found some valuable serum exosomal miRNAs for diagnosis and prognosis of SCI.
Subject(s)
Circulating MicroRNA/genetics , Exosomes/genetics , Spinal Cord Injuries/genetics , Transcriptome , Animals , Biomarkers/blood , Circulating MicroRNA/blood , Exosomes/metabolism , Female , Rats , Rats, Sprague-Dawley , Spinal Cord Injuries/bloodSubject(s)
Carbazoles/therapeutic use , Neuroprotective Agents/therapeutic use , Spinal Cord Injuries/drug therapy , Animals , Female , Motor Activity/drug effects , NAD/metabolism , Neurons/drug effects , Oligodendroglia/drug effects , Rats , Rats, Sprague-Dawley , Spinal Cord/drug effects , Spinal Cord/metabolismSubject(s)
Exosomes/metabolism , MicroRNAs/blood , Spinal Cord Injuries/blood , Spinal Cord Injuries/genetics , Acute Disease , Animals , Down-Regulation/genetics , Exosomes/ultrastructure , Female , Gene Expression Profiling , Gene Ontology , MicroRNAs/genetics , RNA, Untranslated/genetics , RNA, Untranslated/metabolism , Rats, Sprague-Dawley , Reproducibility of Results , Spinal Cord/pathology , Up-Regulation/geneticsSubject(s)
Adaptive Immunity/immunology , Macrophages/immunology , Spinal Cord Injuries/immunology , Transcriptome/genetics , Adaptive Immunity/genetics , Adoptive Transfer/methods , Animals , Down-Regulation/genetics , Down-Regulation/immunology , Female , Inflammation/genetics , Inflammation/immunology , Neuroprotective Agents/immunology , Rats , Rats, Sprague-Dawley , Signal Transduction/genetics , Signal Transduction/immunology , Spinal Cord/immunology , Spinal Cord Injuries/genetics , Toll-Like Receptors/genetics , Transcriptome/immunologyABSTRACT
Previous studies by our group have demonstrated that the transplantation of exogenous platelet-derived growth factor (PDGF)-AA-overexpressing oligodendrocyte progenitor cells (OPCs) promotes tissue repair and recovery of neurological function in a rat model of spinal cord injury (SCI). However, it remains unclear whether treatment with PDGF-AA also affects endogenous oligodendrocytes (OLs) or even neurons, thus promoting further functional recovery after SCI. In the present study, we evaluated the therapeutic potential of PDGF-AA treatment by direct subcutaneous injection of PDGF-AA immediately after SCI. We demonstrated that PDGF-AA injection resulted in increased tissue sparing, myelination and functional recovery in rats following SCI. Further experimentation confirmed that PDGF-AA increased the survival of endogenous OPCs and OLs, and promoted the proliferation of OPCs and their differentiation into OLs. Moreover, PDGF-AA also protected motor neurons from death in the injured spinal cord. These results indicated that PDGF-AA administration may be an effective treatment for SCI.