Previous studies have shown that neuroiflament 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 spec-trometry. In addition, the nerve repair-related neurofilament protein M with the unregulated expression was detected with immunohistochemistry and western blot analysis. Two-dimen-sional gel electrophoresis and mass spectrometry showed that, compared with the sham group, upregulation of protein expression was most signiifcant in the spinal cords of rabbits that had undergone ischemia and 24 hours of reperfusion. Immunohistochemical analysis revealed that neuroiflament 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 neuroiflament protein M expression increased with reperfusion time until it peaked at 24 hours and returned to baseline level after 48 hours. Furthermore, neuroiflament 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.

大鼠肾缺血/再灌注损伤不同时相Fas、FasL蛋白表达及意义

AIM:To study the relationship between the expression of Fas/FasL protein and apoptosis in rats after renal ischemia/reperfusion. METHODS:Establish the models of renal ischemia/reperfusion injury in rat and SABC immunohistochemical methods were used to detect the changes of expression of Fas/FasL protein. Pathomorphological changes in renal ischemia/reperfusion injury were observed.RESULTS:The expression of Fas/FasL proteins was negative in the sham operated group.Fas/FasL proteins were increased in renal in ischemia/reperfusion group,and gradually upregulated with the duration of ischemia or reperfusion and peaked at 72 h of reperfusion.The expression of Fas/FasL proteins was stronger in 60 min ischemia group than 30 min ischemia group and they were mainly expressed in renal tubule.We observed local necrosis and inflammatory cells infiltration around infracted area in ischemia/reperfusion group by HE dyeing methods.And the necrosis area was mainly occurred around proximal convoluted tubule. CONCLUSIONS:These findings suggested that Fas/FasL proteins were over expressed after ischemia/reperfusion injury in rats renal.And Fas/FasL system was involved in the process of renal ischemia/reperfusion injury.