Abstract
ObjectiveHyperbaric oxygen (HBO) preconditioning (HBO-PC) has been testified to have protective effects on spinal cord injury (SCI). However, the mechanisms remain enigmatic. The present study aimed to explore the effects of HBO-PC on primary rat spinal neurons against oxidative injury and oxygen-glucose deprivation (OGD) and the relationship with heat shock proteins (HSPs).MethodsPrimary rat spinal neurons after 7 days of culture were used in this study. HSPs were detected in rat spinal neurons following a single exposure to HBO at different time points by Western blot. Using lactate dehydrogenase release assay and cell counting kit-8 assay, the injuries induced by hydrogen peroxide (H2O2) insult or OGD were determined and compared among neurons treated with HBO-PC with or without HSP inhibitors.ResultsThe results of Western blot showed that HSP27, HSP70 and HSP90 have a slight but not significant increase in primary neurons following HBO exposure. However, HSP32 expression significantly increased and reached highest at 12 h following HBO exposure. HBO-PC significantly increased the cell viability and decreased the medium lactate dehydrogenase content in cultures treated with H2O2 or OGD. Pretreatment with zinc protoporphyrin IX, a specific inhibitor of HSP32, significantly blocked the protective effects of HBO-PC.ConclusionsThese results suggest that HBO-PC could protect rat spinal neurons in vitro against oxidative injury and OGD mostly by up-regulating of HSP32 expression.
Highlights
Spinal cord injury (SCI) is a serious health problem, which may result from surgical operations on the spinal column or thoracoabdominal aorta, or decompression sickness (DCS) associated with sport or commercial diving
Identification of spinal neurons Immunocytochemistry staining of cultures at 7 d in vitro showed that over 90% of the cells counterstained with DAPI were immuno-positive for b-tubulin III, a marker of neurons, which confirmed their neuronal identity (Figure 1)
heat shock proteins (HSPs) expression after Hyperbaric oxygen (HBO) exposure The results of Western blot showed that HSP27, HSP70 and
Summary
Spinal cord injury (SCI) is a serious health problem, which may result from surgical operations on the spinal column or thoracoabdominal aorta, or decompression sickness (DCS) associated with sport or commercial diving. The reported rate of paraplegia or paraparesis after thoracoabdominal aortic aneurysms surgery varies from 3.8% to 16.7% [1,2] and the incidence of injured divers with incomplete recovery from neurological DCS has been reported between 20% and 30% [3] Under those high risk conditions, effective preventive measures are in desperate need to reduce the incidence of SCI. Hyperbaric oxygen (HBO) therapy is the way that a person breathes pure oxygen under a pressure greater than 1 atmosphere absolute in a specially designed chamber [4] It is widely used in the management of various diseases and conditions including DCS, carbon monoxide (CO) poisoning, gas embolism and neurologic diseases [5]. There is still short of evidence about the direct effects of HBO-PC on spinal neurons in vitro and the mechanisms
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