Ischemic preconditioning suppresses apoptosis of rabbit spinal neurocytes by inhibiting ASK1-14-3-3 dissociation
Ischemic preconditioning suppresses apoptosis of rabbit spinal neurocytes by inhibiting ASK1-14-3-3 dissociation
复制标题
缺血预处理通过抑制 ASK1-14-3-3 解离抑制兔脊髓神经细胞凋亡
DOI：
10.1016/j.neulet.2008.06.037
复制
发表时间：
2008-08-29
影响因子：
2.5
通讯作者：
Yin, Guoyong
中科院分区：
文献类型：
--
作者：
Yang, Chengwei;Ren, Yongxin;Yin, Guoyong
短暂的亚致死性缺血后再灌流(缺血预适应，IPC)阻止随后长时间缺血的致死效应的机制尚不清楚。采用兔脊髓缺血损伤模型，设计了一项完全随机的对照研究，以研究IPC的作用。成年新西兰大白兔24只，随机分为3组(每组8只)：I组：假手术组；II组：缺血再灌注(I/R)组；采用肾下主动脉阻断30min复制脊髓缺血模型。损伤后，II组兔进行30min、2 h或8 h的再灌注。III组动物进行3个循环，每个循环5分钟，然后再灌流5分钟，然后接受30分钟的缺血。我们先前报道，ASK1与14-3-3之间的关系在调节缺血再灌注脊髓损伤中起着重要作用。为了评估缺血预适应对损伤脊髓的影响，我们检测了脊髓组织形态的变化，ASK1介导的信号通路关键成员的激活，以及ASK1和14-3-3之间的联系。用苏木精-伊红(H&E)染色和电子显微镜观察脊髓的形态变化。免疫印迹法检测ASK1、JNK和p38的磷酸化水平。免疫共沉淀实验分析ASK1与14-3-3的相关性。结果表明，缺血预适应对ASK1/14-3-3解离所致脊髓损伤有保护作用。与II组相比，III组脊髓神经元胞体肿胀和出血明显减少，神经细胞凋亡程度减轻。(C)2008爱思唯尔爱尔兰有限公司。保留所有权利。
The mechanism by which a brief episode of sublethal ischemia followed by reperfusion (ischemic preconditioning, IPC) prevents the lethal effects of subsequent periods of prolonged ischemia, are poorly understood. A completely randomized, controlled study was designed to study the effect of IPC using a rabbit model of ischemic spinal cord injury. Twenty-four white adult New England rabbits were randomly assigned to one of 3 groups (n = 8 per group); the groups were assigned as follows: Group I: sham-operation group, Group II: ischemic reperfusion (I/R) group, and Group III: ischemic preconditioning group. Spinal cord ischemia was induced by introducing an infra renal aortic cross-clamp for 30 min. Following injury, rabbits were subjected to 30 min, 2 h, or 8 h of reperfusion in Group II. In Group III, subjects underwent three cycles, 5 min each, of ischemia followed by 5 min of reperfusion, before receiving 30 min of ischemia. We previously reported that the association between ASK1 (apoptosis signal-regulating kinase 1) and 14-3-3 played an important role in regulating ischemia)reperfusion spinal cord injuries. To evaluate the effect of ischemic preconditioning in injured spinal cords, we examined alterations in spinal tissue morphology, activation of key members of the ASK1-mediated signaling pathway, and the association between ASK1 and 14-3-3. Changes in spinal cord morphology were observed with hematoxylin and eosin (H&E) staining and electron microscopy. The phosphorylation levels of ASK1, JNK, and p38 were assessed by immunoblot analysis. The association between ASK1 and 14-3-3 was analyzed by co-immunoprecipitation experiments. We observed that swelling of the neurocyte bodies and hemorrhage of the spinal cord were dramatically decreased in Group III compared to Group II. In addition, the degree of apoptosis among neurocytes was reduced in Group III compared to Group II. Finally, the phosphorylation of ASK1, JNK, p38 and the dissociation of ASK1 from 14-3-3 were dramatically decreased in Group III compared with Group II. These results indicate that ischemic preconditioning may have a protective affect against ASK1/14-3-3 dissociation-induced spinal cord injuries. (c) 2008 Elsevier Ireland Ltd. All rights reserved.