Yamauchi Takashi, Sakurai Masahiro, Abe Koji, Matsumiya Goro, Sawa Yoshiki
Department of Cardiovascular Surgery, Osaka University Graduate School of Medicine, 2-2 Yamadaoka, Suita, Osaka, 565-0871, Japan.
Brain Res. 2007 Sep 12;1169:24-33. doi: 10.1016/j.brainres.2007.06.093. Epub 2007 Jul 31.
Delayed paraplegia after operation of the thoracic aorta is considered to be related to vulnerability of motor neurons to ischemia. Recently, endoplasmic reticulum (ER) stress has been reported to participate in neuronal cell death. In the present study, we investigate the expression of ER stress-related molecules and discuss the relationship between neuronal vulnerability and ER stress after transient ischemia in the spinal cord.
A rabbit spinal cord ischemia model was generated using a balloon catheter. In this model, spinal motor neurons show selectively delayed neuronal death whereas other spinal neuron, such as interneurons, survive. Immunohistochemical analysis and Western blotting for ER stress-related molecules, including phosphorylated eukaryotic initiation factor 2 alpha (p-eIF2alpha), activating transcription factor 4 (ATF4), glucose-regulated protein 78 (GRP78) and inositol-requiring ER transmembrane RNAse alpha isoform (IRE1alpha), were examined.
P-eIF2alpha, which inhibits protein synthesis and modulates ER stress, was induced only in interneurons after 6 h of reperfusion. ATF4, which is specifically activated by PERK-eIF2alpha, was induced only in interneurons between 6 h and 1 day after reperfusion. GRP78 was induced strongly both in interneurons and motor neurons at an early stage of reperfusion, but prolonged expression was observed only in interneurons. IRE1alpha, which is supposed to transduce an ER stress-related death signal, was expressed more strongly and over a more prolonged period in motor neurons.
These results indicate that the vulnerability of motor neurons in the spinal cord might be partially attributed to an ER stress response to transient ischemia.
胸主动脉手术后发生的迟发性截瘫被认为与运动神经元对缺血的易损性有关。最近,有报道称内质网(ER)应激参与神经元细胞死亡。在本研究中,我们调查了ER应激相关分子的表达,并探讨脊髓短暂缺血后神经元易损性与ER应激之间的关系。
使用球囊导管建立兔脊髓缺血模型。在该模型中,脊髓运动神经元表现出选择性延迟性神经元死亡,而其他脊髓神经元,如中间神经元,则存活。对ER应激相关分子进行免疫组织化学分析和蛋白质印迹检测,包括磷酸化真核起始因子2α(p-eIF2α)、活化转录因子4(ATF4)、葡萄糖调节蛋白78(GRP78)和肌醇需求性ER跨膜RNA酶α亚型(IRE1α)。
抑制蛋白质合成并调节ER应激的p-eIF2α仅在再灌注6小时后在中间神经元中被诱导。由PERK-eIF2α特异性激活的ATF4仅在再灌注后6小时至1天之间在中间神经元中被诱导。GRP78在再灌注早期在中间神经元和运动神经元中均被强烈诱导,但仅在中间神经元中观察到其持续表达。推测可转导ER应激相关死亡信号的IRE1α在运动神经元中表达更强且持续时间更长。
这些结果表明,脊髓运动神经元的易损性可能部分归因于对短暂缺血的ER应激反应。