Previous studies have shown that
neurofilament protein M expression is upregulated in the early stage of
spinal cord ischemia/
reperfusion injury, indicating that this
protein may play a role in the injury process. In the present study, we compared
protein expression in spinal cord tissue of rabbits after 25 minutes of
ischemia followed by 0, 12, 24, or 48 hours of reperfusion with that of
sham operated rabbits, using proteomic two-dimensional gel electrophoresis and mass spectrometry. In addition, the nerve repair-related
neurofilament protein M with the unregulated expression was detected with immunohistochemistry and western blot analysis. Two-dimensional gel electrophoresis and mass spectrometry showed that, compared with the
sham group, upregulation of
protein expression was most significant in the spinal cords of rabbits that had undergone
ischemia and 24 hours of reperfusion. Immunohistochemical analysis revealed that
neurofilament protein M was located in the membrane and cytoplasm of neuronal
soma and axons at each time point after injury. Western blot analysis showed that
neurofilament protein M expression increased with reperfusion time until it peaked at 24 hours and returned to baseline level after 48 hours. Furthermore,
neurofilament protein M is phosphorylated under oxidative stress, and expression changes were parallel for the phosphorylated and non-phosphorylated forms.
Neurofilament protein M plays an important role in
spinal cord ischemia/
reperfusion injury, and its functions are achieved through oxidative phosphorylation.