Selective release of calpain produced αII-spectrin (α-fodrin) breakdown products by acute neuronal cell death
Selective release of calpain produced αII-spectrin (α-fodrin) breakdown products by acute neuronal cell death
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DOI：
10.1515/bc.2002.082
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发表时间：
2002-05-01
影响因子：
3.7
通讯作者：
Wang, KKW
中科院分区：
文献类型：
--
作者：
Dutta, S;Chiu, YC;Wang, KKW
钙蛋白酶的激活导致[α]II血影蛋白([α]fodrin)的分解，这是一种神经元细胞骨架蛋白，已在各种体外和体内神经元损伤模型中检测到。在这项研究中，发现150 kDa的血影蛋白降解产物(SBDP150)被钙通道开放剂麦托毒素(MTX)处理的SHSY5Y细胞释放到细胞条件培养液中。使用SBDP150特异性多克隆抗体可以在免疫印迹上很容易地定量SBDP150的释放。SBDP150的升高也与细胞死亡呈时间依赖关系。选择性钙蛋白酶抑制剂MDL28170是唯一一种可显著降低MTX诱导的SBDP150释放的蛋白酶抑制剂。兴奋性毒素(NMDA和海人藻酸)刺激的小脑颗粒神经元细胞培养上清液中SBDP150也显著增加，而NMDA受体拮抗剂R()-3-(2-碳哌嗪-4-基)丙基-1-膦酸(CPP)和MDL28170可减弱这一作用。此外，大脑皮层培养的低氧/低血糖挑战也导致SBDP150被释放到媒体中。这些结果支持一种理论，即基于抗体的细胞条件培养液中SBDP150的检测可以用于量化神经细胞的损伤。此外，SBDP150有可能在临床环境中作为急性神经元损伤的替代生物标志物。
Activation of calpain results in the breakdown of [alpha] II spectrin ([alpha]fodrin), a neuronal cytoskeleton protein, which has previously been detected in various in vitro and in vivo neuronal injury models. In this study, a 150 kDa spectrin breakdown product (SBDP150) was found to be released into the cellconditioned media from SHSY5Y cells treated with the calcium channel opener maitotoxin (MTX). SBDP150 release can be readily quantified on immunoblot using an SBDP150- specific polyclonal antibody. Increase of SBDP150 also correlated with cell death in a timedependent manner. MDL28170, a selective calpain inhibitor, was the only protease inhibitor tested that significantly reduced MTXinduced SBDP150 release. The cellconditioned media of cerebellar granule neurons challenged with excitotoxins (NMDA and kainate) also exhibited a significant increase of SBDP150 that was attenuated by pretreatment with an NMDA receptor antagonist, R()-3-(2-carbopiperazine-4-yl)propyl-1- phosphonic acid (CPP), and MDL28170. In addition, hypoxic/hypoglycemic challenge of cerebrocortical cultures also resulted in SBDP150 liberation into the media. These results support the theory that an antibody based detection of SBDP150 in the cellconditioned media can be utilized to quantify injury to neural cells. Furthermore, SBDP150 may potentially be used as a surrogate biomarker for acute neuronal injury in clinical settings.