Early processing of Bid and caspase-6,-8,-10,-14 in the canine brain during cardiac arrest and resuscitation
Early processing of Bid and caspase-6,-8,-10,-14 in the canine brain during cardiac arrest and resuscitation
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DOI：
10.1016/j.expneurol.2004.05.020
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发表时间：
2004-10-01
影响因子：
5.3
通讯作者：
Krajewski, S
中科院分区：
文献类型：
--
作者：
Krajewska, M;Rosenthal, RE;Krajewski, S
采用临床相关的狗短暂性全脑缺血伴心脏骤停复苏模型，研究凋亡调节蛋白的表达和蛋白水解过程。缺血再灌注损伤后，海马组织中Bcl-2家族促凋亡蛋白Bcl-X-s和Bak表达增加，同时BcI-X-L和Bcl-2蛋白水解。此外，在大脑的这一区域发现了Bid的双相分裂，tBid-p11在心脏骤停后10分钟内早期生成，随后在30分钟的再灌注内生成tBid-p15，这与该促凋亡蛋白的激活一致。此外，心脏骤停和复苏诱导了caspase-3激活之前的caspase-6、-8、-10和-14的早期再灌注依赖性蛋白水解过程。使用抗体的免疫组织化学分析，优先识别加工的caspase-3， -6， -8和-10，提供了这些蛋白酶在大脑缺血敏感区域的神经元和胶质细胞中的时间依赖性激活的证据。总之，在心脏骤停和复苏引起的缺血性脑损伤的临床相关模型中，凋亡调节蛋白发生了极其快速的细胞选择性加工。缺血诱导后犬海马中Bid的早期裂解和32kda前caspase-14的快速耗竭表明钙蛋白酶参与了这些蛋白的加工。本研究中calpain I体外切割重组小鼠caspase-14的实验证实了这一假设，并进一步暗示不同蛋白酶家族之间的相互作用参与了缺血性神经细胞死亡的病理生理过程。(C) 2004爱思唯尔公司版权所有。
A clinically relevant model of transient global brain ischemia involving cardiac arrest followed by resuscitation in dogs was utilized to study the expression and proteolytic processing of apoptosis-regulatory proteins. In the hippocampus, an increase in pro-apoptotic Bcl-2 family proteins Bcl-X-s and Bak was detected, concomitant with proteolysis of BcI-X-L and Bcl-2, following ischemia-reperfusion injury. Also, biphasic cleavage of Bid was found in this region of the brain, with early generation of tBid-p11 within 10 min of cardiac arrest, followed by generation of tBid-p15 within 30-min reperfusion, consistent with activation of this pro-apoptotic protein. In addition, cardiac arrest and resuscitation induced early, reperfusion-dependent proteolytic processing of pro-caspase-6, -8, -10, and -14, which preceded caspase-3 activation. Immunohistochemical analysis using antibodies, which preferentially recognize processed caspase-3, -6, -8, and -10, provided evidence of time-dependent activation of these proteases in both neurons and glia in ischemia-sensitive regions of the brain.In conclusion, extremely rapid, cell-selective processing of apoptosis-regulatory proteins occurs in a clinically relevant model of ischemic brain injury caused by cardiac arrest and resuscitation. The early cleavage of Bid and rapid depletion of 32-kDa pro-caspase-14 from the canine hippocampus after induction of ischemia suggests the involvement of calpains in the processing of these proteins. Demonstration of in vitro cleavage of recombinant mouse caspase-14 by calpain I in the present study lends support to this hypothesis, further implicating crosstalk between different protease families in the pathophysiology of ischemic neural cell death. (C) 2004 Elsevier Inc. All rights reserved.