scholarly journals Transcriptomic Screening of Microvascular Endothelial Cells Implicates Novel Molecular Regulators of Vascular Dysfunction after Spinal Cord Injury

2008 ◽  
Vol 28 (11) ◽  
pp. 1771-1785 ◽  
Author(s):  
Richard L Benton ◽  
Melissa A Maddie ◽  
Christopher A Worth ◽  
Edward T Mahoney ◽  
Theo Hagg ◽  
...  
Keyword(s):  
Spinal Cord ◽  
Spinal Cord Injury ◽  
Vascular Dysfunction ◽  
Fluorescein Isothiocyanate ◽  
Microvascular Dysfunction ◽  
Cell Counting ◽  
Protein Levels ◽  
Cord Injury ◽  
Biochemical Analyses ◽  
High Degree

Microvascular dysfunction is a critical pathology that underlies the evolution of secondary injury mechanisms after traumatic spinal cord injury (SCI). However, little is known of the molecular regulation of endothelial cell (EC) plasticity observed acutely after injury. One reason for this is the relative lack of methods to quickly and efficiently obtain highly enriched spinal microvascular ECs for high-throughput molecular and biochemical analyses. Adult C57BI/6 mice received an intravenous injection of fluorescein isothiocyanate (FITC)-conjugated Lycopersicon esculentum lectin, and FITC-lectin bound spinal microvessels were greatly enriched by fluorescence-activated cell sorter (FACS) purification. This technique allows for rapid (< 1.5 h postmortem) isolation of spinal cord microvascular ECs (smvECs). The results from cell counting, reverse-transcription polymerase chain reaction (RT-PCR), and western blot analyses show a high degree of EC enrichment at mRNA and protein levels. Furthermore, a focused EC biology microarray analysis identified multiple mRNAs dramatically increased in the EC compartment 24 h after SCI, which is a time point associated with the pathologic loss of spinal vasculature. These included thrombospondin-1, CCL5/RANTES, and urokinase plasminogen activator, suggesting they may represent targets for therapeutic intervention. Furthermore, these novel methodologic approaches will likely facilitate the discovery of molecular regulators of endothelial dysfunction in a variety of central nervous system (CNS) disorders including stroke and other neurodegenerative diseases having a vascular component.

scholarly journals D-dopachrome tautomerase activates COX2/PGE2 pathway of astrocytes to mediate inflammation following spinal cord injury

2021 ◽  
Vol 18 (1) ◽  
Author(s):  
Huiyuan Ji ◽  
Yuxin Zhang ◽  
Chen Chen ◽  
Hui Li ◽  
Bingqiang He ◽  
...  
Keyword(s):  
Spinal Cord ◽  
Spinal Cord Injury ◽  
Signal Pathways ◽  
Post Injury ◽  
Dopachrome Tautomerase ◽  
Protein Levels ◽  
Cytokines And Chemokines ◽  
Cord Injury ◽  
Mitogen Activated Protein ◽  
Spinal Cord Contusion

Abstract Background Astrocytes are the predominant glial cell type in the central nervous system (CNS) that can secrete various cytokines and chemokines mediating neuropathology in response to danger signals. D-dopachrome tautomerase (D-DT), a newly described cytokine and a close homolog of macrophage migration inhibitory factor (MIF) protein, has been revealed to share an overlapping function with MIF in some ways. However, its cellular distribution pattern and mediated astrocyte neuropathological function in the CNS remain unclear. Methods A contusion model of the rat spinal cord was established. The protein levels of D-DT and PGE2 synthesis-related proteinase were assayed by Western blot and immunohistochemistry. Primary astrocytes were stimulated by different concentrations of D-DT in the presence or absence of various inhibitors to examine relevant signal pathways. The post-injury locomotor functions were assessed using the Basso, Beattie, and Bresnahan (BBB) locomotor scale. Results D-DT was inducibly expressed within astrocytes and neurons, rather than in microglia following spinal cord contusion. D-DT was able to activate the COX2/PGE2 signal pathway of astrocytes through CD74 receptor, and the intracellular activation of mitogen-activated protein kinases (MAPKs) was involved in the regulation of D-DT action. The selective inhibitor of D-DT was efficient in attenuating D-DT-induced astrocyte production of PGE2 following spinal cord injury, which contributed to the improvement of locomotor functions. Conclusion Collectively, these data reveal a novel inflammatory activator of astrocytes following spinal cord injury, which might be beneficial for the development of anti-inflammation drug in neuropathological CNS.

Recurrent autonomic dysreflexia exacerbates vascular dysfunction after spinal cord injury

2010 ◽  
Vol 10 (12) ◽  
pp. 1108-1117 ◽  
Author(s):  
Nima Alan ◽  
Leanne M. Ramer ◽  
Jessica A. Inskip ◽  
Saeid Golbidi ◽  
Matt S. Ramer ◽  
...  
Keyword(s):  
Spinal Cord ◽  
Spinal Cord Injury ◽  
Vascular Dysfunction ◽  
Autonomic Dysreflexia ◽  
Cord Injury

Expressions of Nogo-66 Receptor at mRNA and Protein Levels After Spinal Cord Injury in Rats

2012 ◽  
Vol 12 (9) ◽  
pp. S153
Author(s):  
Huilin Yang ◽  
Jibin Wu ◽  
Tiansi Tang ◽  
Muhammad Z. Moral ◽  
Liang Chen ◽  
...  
Keyword(s):  
Spinal Cord ◽  
Spinal Cord Injury ◽  
Protein Levels ◽  
Cord Injury

scholarly journals Association Between Mobility Mode and C-Reactive Protein Levels in Men With Chronic Spinal Cord Injury

2008 ◽  
Vol 89 (4) ◽  
pp. 726-731 ◽  
Author(s):  
Leslie R. Morse ◽  
Kelly Stolzmann ◽  
Hiep P. Nguyen ◽  
Nitin B. Jain ◽  
Cara Zayac ◽  
...  
Keyword(s):  
Spinal Cord ◽  
Spinal Cord Injury ◽  
C Reactive Protein ◽  
Chronic Spinal Cord Injury ◽  
Protein Levels ◽  
Reactive Protein ◽  
Cord Injury

C-terminal domain of tetanus toxin changes the apoptosis- and autophagy-related protein levels following spinal cord injury in rat brain

2017 ◽  
Vol 108 ◽  
pp. S89
Author(s):  
Murat Celal Sözbilen ◽  
Murat Öztürk ◽  
Gizem Kaftan ◽  
Taner Dağcı ◽  
Halit Özyalçın ◽  
...  
Keyword(s):  
Spinal Cord ◽  
Spinal Cord Injury ◽  
Rat Brain ◽  
Tetanus Toxin ◽  
Related Protein ◽  
Protein Levels ◽  
Terminal Domain ◽  
Cord Injury

scholarly journals Neuroprotective effects of electro-acupuncture in spinal cord injury rats via up-regulation of DUSP14

2021 ◽  
Vol 18 (9) ◽  
pp. 1831-1837
Author(s):  
Junfeng Zhang ◽  
Yaochi Wu ◽  
Jingjie Xu ◽  
Shenghong Zhang ◽  
Shisheng Li
Keyword(s):  
Spinal Cord ◽  
Spinal Cord Injury ◽  
Neuroprotective Effects ◽  
Strand Breaks ◽  
Over Expression ◽  
Protein Levels ◽  
Cord Injury ◽  
Dna Strand ◽  
Factor Α ◽  
Intraspinal Injection

Purpose: To study the effect and mechanism of action of electro-acupuncture (EA) on nerve regeneration by analyzing the behavior, inflammation and cell death in spinal cord injury (SCI) rat model. Methods: SCI model was established according to Allen’s falling strike method. Electroacupuncture was performed on Jiaji (EX-B2)/Mingmen (GV4) acupoint with a 1 mA current intermittent wave at a frequency of 2Hz for 20 min daily. Interleukin (IL-6) and tumor necrosis factor-α (TNF-α) levels were measured using ELISA kits. Apoptosis-induced DNA strand breaks were evaluated by TUNEL assay while relative mRNA expression was assessed by quantitative real-time polymerase chain reaction (qRT-PCR). Protein levels were measured by western blot. Results: Relative mRNA and protein expressions of DUSP14 decreased in SCI rats with time but increased by EA treatment. Further, partial locomotor functional recovery was presented in SCI rats by EA treatment. Moreover, intraspinal injection of DUSP14 over-expression viral supernatants/EA treatment ameliorated inflammation and apoptosis in SCI rats. Meanwhile, the protein levels of NF-κB p65 (nucleus) and phosphorylated TGF-activated kinase 1 (p-TAK1) increased in SCI rats following EA treatment but were decreased by EA treatment and intraspinal injection of DUSP14 over-expression viral supernatants. Conclusion: EA acupoint treatment exerts neuroprotective effects in SCI rats via the reduction of inflammation and apoptosis, and induction of DUSP14.

scholarly journals LncGBP9/miR-34a axis drives macrophages toward a phenotype conducive for spinal cord injury repair via STAT1/STAT6 and SOCS3

2020 ◽  
Author(s):  
Jiahui Zhou ◽  
Zhiyue Li ◽  
Tianding Wu ◽  
Qun Zhao ◽  
Qiancheng Zhao ◽  
...  
Keyword(s):  
Spinal Cord ◽  
Spinal Cord Injury ◽  
Macrophage Polarization ◽  
Cytokine Signaling ◽  
Secondary Injury ◽  
Protein Levels ◽  
M2 Polarization ◽  
Cord Injury ◽  
Socs3 Expression ◽  
M1 Phenotype

Abstract Background: Acute spinal cord injury (SCI) could cause mainly two types of pathological sequelae, the primary mechanical injury, and the secondary injury. The macrophage in SCI are skewed toward the M1 phenotype that might cause the failure to post-SCI repair. Methods: SCI model was established in Balb/c mice, and the changes in macrophage phenotypes after SCI were monitored. Bioinformatic analyses were performed to select factors that might regulate macrophage polarization after SCI. Mouse bone marrow-derived macrophages ( BMDMs ) were isolated, identified, and induced for M1 or M2 polarization; the effects of lncRNA guanylate binding protein-9 (lnc GBP9 ) and suppressor of cytokine signaling 3 (SOCS 3) on macrophages polarization were examined in vitro and in vivo . The predicted miR-34a binding to lncGBP9 and SOCS3 was validated; the dynamic effects of lncGBP9 and miR-34a on SOCS3, signal transducer and activator of transcription 1 (STAT 1) /STAT6 signaling, and macrophage polarization were examined. Finally, we investigated whether STAT6 could bind the miR-34a promoter to activate its transcription. Results: In SCI Balb/c mice, macrophage skewing toward M1 phenotypes was observed after SCI. In M1 macrophages, lncGBP9 silencing significantly decreased p-STAT1 and SOCS3 expression and protein levels, as well as the production of Interleukin (IL)-6 and IL-12; in M2 macrophages, lncGBP9 overexpression increased SOCS3 mRNA expression and protein levels while suppressed p-STAT6 levels and the production of IL-10 and transforming growth factor-beta 1 (TGF-β1 ) , indicating that lncGBP9 overexpression promotes the M1 polarization of macrophages. In lncGBP9-silenced SCI mice, the M2 polarization was promoted on day 28 after the operation, further indicating that lncGBP9 silencing revised the predominance of M1 phenotype at the late stage of secondary injury after SCI, therefore improving the repair after SCI. IncGBP9 competed with SOCS3 for miR-34a binding to counteract miR-34a-mediated suppression on SOCS3 and then modulated STAT1/STAT6 signaling and the polarization of macrophages. STAT6 bound the promoter of miR-34a to activate its transcription. Conclusions: In macrophages, lncGBP9 sponges miR-34a to rescue SOCS3 expression, therefore modulating macrophage polarization through STAT1/STAT6 signaling. STAT6 bound the promoter of miR-34a to activate its transcription, thus forming two different regulatory loops to modulate the phenotype of macrophages after SCI.

scholarly journals Transplantation of sh-miR-199a-5p-Modified Olfactory Ensheathing Cells Promotes the Functional Recovery in Rats with Contusive Spinal Cord Injury

2020 ◽  
Vol 29 ◽  
pp. 096368972091617 ◽  
Author(s):  
Zhengchao Gao ◽  
Yingjie Zhao ◽  
Xijing He ◽  
Zikuan Leng ◽  
Xiaoqian Zhou ◽  
...  
Keyword(s):  
Spinal Cord ◽  
Spinal Cord Injury ◽  
Functional Recovery ◽  
Diffusion Tensor ◽  
Olfactory Ensheathing Cells ◽  
Luciferase Reporter ◽  
Therapeutic Effects ◽  
Protein Levels ◽  
Cord Injury ◽  
Ensheathing Cells

MicroRNAs (miRNAs) function as gene expression switches, and participate in diverse pathophysiological processes of spinal cord injury (SCI). Olfactory ensheathing cells (OECs) can alleviate pathological injury and facilitate functional recovery after SCI. However, the mechanisms by which OECs restore function are not well understood. This study aims to determine whether silencing miR-199a-5p would enhance the beneficial effects of the OECs. In this study, we measured miR-199a-5p levels in rat spinal cords with and without injury, with and without OEC transplants. Then, we transfected OECs with the sh-miR-199a-5p lentiviral vector to reduce miR-199a-5p expression and determined the effects of these OECs in SCI rats by Basso–Beattie–Bresnahan (BBB) locomotor scores, diffusion tensor imaging (DTI), and histological methods. We used western blotting to measure protein levels of Slit1, Robo2, and srGAP2. Finally, we used the dual-luciferase reporter assay to assess the relationship between miR-199-5p and Slit1, Robo2, and srGAP2 expression. We found that SCI significantly increased miR-199a-5p levels ( P < 0.05), and OEC transplants significantly reduced miR-199a-5p expression ( P < 0.05). Knockdown of miR-199a-5p in OECs had a better therapeutic effect on SCI rats, indicated by higher BBB scores and fractional anisotropy values on DTI, as well as histological findings. Reducing miR-199a-5p levels in transplanted OECs markedly increased spinal cord protein levels of Slit1, Robo2, and srGAP2. Our results demonstrated that transplantation of sh-miR-199a-5p-modified OECs promoted functional recovery in SCI rats, suggesting that miR-199a-5p knockdown was more beneficial to the therapeutic effects of OEC transplants. These findings provided new insights into miRNAs-mediated therapeutic mechanisms of OECs, which helps us to develop therapeutic strategies based on miRNAs and optimize cell therapy for SCI.

scholarly journals Current status of acute spinal cord injury pathophysiology and emerging therapies: promise on the horizon

2008 ◽  
Vol 25 (5) ◽  
pp. E2 ◽  
Author(s):  
James W. Rowland ◽  
Gregory W. J. Hawryluk ◽  
Brian Kwon ◽  
Michael G. Fehlings
Keyword(s):  
Spinal Cord ◽  
Spinal Cord Injury ◽  
Apoptotic Cell ◽  
Vascular Dysfunction ◽  
Mechanical Injury ◽  
Current Status ◽  
Emerging Therapies ◽  
Cord Injury ◽  
Central Myelin ◽  
Do So

This review summarizes the current understanding of spinal cord injury pathophysiology and discusses important emerging regenerative approaches that have been translated into clinical trials or have a strong potential to do so. The pathophysiology of spinal cord injury involves a primary mechanical injury that directly disrupts axons, blood vessels, and cell membranes. This primary mechanical injury is followed by a secondary injury phase involving vascular dysfunction, edema, ischemia, excitotoxicity, electrolyte shifts, free radical production, inflammation, and delayed apoptotic cell death. Following injury, the mammalian central nervous system fails to adequately regenerate due to intrinsic inhibitory factors expressed on central myelin and the extracellular matrix of the posttraumatic gliotic scar. Regenerative approaches to block inhibitory signals including Nogo and the Rho-Rho–associated kinase pathways have shown promise and are in early stages of clinical evaluation. Cell-based strategies including using neural stem cells to remyelinate spared axons are an attractive emerging approach.

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