Objectives: Necroptosis, a form of regulated necrosis, might be a potential mechanism of delayed paraplegia; therefore, its role in transient spinal cord ischaemia was investigated by immunohistochemical analysis of necroptosis related protein receptor interacting protein kinase (RIP) 1, RIP3, and cellular inhibitor of apoptosis protein (cIAP) 1/2. Methods: This study used rabbit normothermic (n = 24) and hypothermic (n = 24) transient spinal cord ischaemia models and sham controls (n = 6). Neurological function was assessed according to a modified Tarlov score at 8 h, 1, 2, and 7 days after reperfusion (n = 6 each). Morphological changes in the spinal cord were examined using haematoxylin and eosin staining in the sham, 2, and 7 day groups. Western blot and histochemical analyses of RIP1, RIP3, and cIAP1/2, and double label fluorescent immunocytochemical studies of RIP3 and cIAP1/2 were performed at 8 h, 1, and 2 days after reperfusion (n = 6 each). Results: There were significant differences in neurological function between the normothermic and hypothermic groups (median scores 0 and 5 at 7 days, p =.023). In the normothermic group, most motor neurons were lost seven days after reperfusion (p =.046 compared with sham), but they were preserved in the hypothermic group. Western blot analysis revealed the upregulation of RIP1, RIP3, and cIAP1/2 at 8 h in the normothermic group (RIP1, p =.032; RIP3, p <.001; cIAP1/2, p =.041 compared with sham), and the overexpression of RIP3 was prolonged for two days. In the hypothermic group, the expression of these proteins was not observed. The double label fluorescent immunocytochemical study revealed the induction of RIP3 and cIAP1/2 in the same motor neurons. Conclusions: These data suggest that transient normothermic ischaemia induces necroptosis, a potential factor in delayed motor neuron death, and that hypothermia may inhibit necroptosis.
|Number of pages||10|
|Journal||European Journal of Vascular and Endovascular Surgery|
|Publication status||Published - May 2020|
- Delayed paraplegia
- Spinal cord ischaemia
ASJC Scopus subject areas
- Cardiology and Cardiovascular Medicine