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Status |
Public on May 18, 2022 |
Title |
Hyperbaric oxygen therapy after mid-cervical spinal contusion injury |
Organism |
Rattus norvegicus |
Experiment type |
Expression profiling by array
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Summary |
Hyperbaric oxygen (HBO) therapy is frequently used to treat peripheral wounds or decompression sickness. Evidence suggests that HBO therapy can provide neuroprotection and has an anti-inflammatory impact after neurologic injury including spinal cord injury (SCI). Our primary purpose was to conduct an unbiased, genome-wide screening of mRNA expression changes in the injured spinal cord after HBO therapy. A mRNA gene array was used to evaluate samples taken from the contused region of the spinal cord following a lateralized mid-cervical contusion injury in adult female rats. HBO therapy consisted of daily, 1-hour sessions (3.0 ATA, 100% O2) initiated on the day of SCI. Gene set enrichment analyses indicated that HBO upregulated genes in pathways associated with electron transport, mitochondrial function, and oxidative phosphorylation, and downregulation genes in pathways associated with inflammation (including cytokines and NF-B) and apoptotic signaling. In a separate cohort, spinal cord histology was performed to verify whether the HBO treatment impacted neuronal cell counts or inflammatory markers. Compared to untreated rats, there was increased NeuN positive cells in the spinal cord of HBO treated rats (p=0.007). Further, staining for anti-ionized calcium binding adaptor protein (Iba-1, a microglial marker) was reduced after HBO (p=0.028). We conclude that HBO therapy, initiated shortly after SCI and continued for 10-days, can alter the molecular signature of the lesioned spinal cord in a manner consistent with an anti-inflammatory and neuroprotective impact.
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Overall design |
A mRNA gene array was used to evaluate samples taken from the contused region of the spinal cord following a lateralized mid-cervical contusion injury in adult female rats. HBO therapy consisted of daily, 1-hour sessions (3.0 ATA, 100% O2) initiated on the day of SCI. All procedures were approved by the University of Florida Institutional Animal Care and Use Committee. Rats were housed in an Assessment and Accreditation of Laboratory Animal Care (AAALAC)-accredited facility with a 12:12 light/dark cycle with ad libitum access to rodent chow and water. Adult female Sprague-Dawley rats (200-250g, Harlan Laboratories, Indianapolis, IN, USA) were studied. Rats (n = 4/group) were injected intraperitoneally with beuthanasia solution. After the loss of the hind-limb withdrawal reflex, cervical spinal tissues (C3-C5) were harvested, placed into RNA Later (Life Technologies, Carlsbad, CA, USA), and stored at -80°C. RNA extraction was performed using TRIzol and isolated total RNA was purified using an RNeasy Mini kit (Qiagen, Valencia, CA), according to manufacturer’s instructions. The resulting quantity and purity of total RNA was tested through absorbance spectrophotometry at 230, 260 and 280 nm. For microarray analysis, RNA samples were sent to the Boston University Medical Center Microarray Core Facility for analysis using the Affymetrix Rat Gene Array 2.0ST. Note: the gene array work was completed on the following groups: naive (spinal intact, no prior surgery); sham (sham surgery, no spinal cord injury); cSCI (spinal contusion, no further treatment), HBOT1d (spinal cord injury followed by HBO treatment only on day 1 post-injury), HBOT10d (spinal cord injury followed by HBO treatment for 10 days). Only the cSCI and HBOT10d groups are included in the accompanying manuscript.
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Contributor(s) |
Fuller D |
Citation(s) |
35152735 |
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Submission date |
Oct 09, 2021 |
Last update date |
May 20, 2022 |
Contact name |
Boston University Microarray and Sequencing Resource |
E-mail(s) |
msrdata@bu.edu
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Organization name |
Boston University
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Department |
Microarray and Sequencing Resource
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Street address |
72 East Concord Street, E631
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City |
Boston |
State/province |
MA |
ZIP/Postal code |
02118 |
Country |
USA |
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Platforms (1) |
GPL17799 |
[RaGene-2_0-st] Affymetrix Rat Gene 2.0 ST Array [ragene20st_Rn_ENTREZG ENTREZG_17.0.0] |
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Samples (20)
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GSM5620469 |
spinal cord cells, naive 4 |
GSM5620470 |
spinal cord cells, sham 1 |
GSM5620471 |
spinal cord cells, sham 2 |
GSM5620472 |
spinal cord cells, sham 3 |
GSM5620473 |
spinal cord cells, sham 4 |
GSM5620474 |
spinal cord cells, cSCI 1 |
GSM5620475 |
spinal cord cells, cSCI 2 |
GSM5620476 |
spinal cord cells, cSCI 3 |
GSM5620477 |
spinal cord cells, cSCI 4 |
GSM5620478 |
spinal cord cells, HBOT 1d 1 |
GSM5620479 |
spinal cord cells, HBOT 1d 2 |
GSM5620480 |
spinal cord cells, HBOT 1d 3 |
GSM5620481 |
spinal cord cells, HBOT 1d 4 |
GSM5620482 |
spinal cord cells, HBOT 10d 1 |
GSM5620483 |
spinal cord cells, HBOT 10d 2 |
GSM5620484 |
spinal cord cells, HBOT 10d 3 |
GSM5620485 |
spinal cord cells, HBOT 10d 4 |
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Relations |
BioProject |
PRJNA769951 |
Supplementary file |
Size |
Download |
File type/resource |
GSE185600_RAW.tar |
173.2 Mb |
(http)(custom) |
TAR (of CEL) |
Processed data included within Sample table |
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