Tributyltin-induced cell death is mediated by calpain in PC12 cells
Tributyltin-induced cell death is mediated by calpain in PC12 cells
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DOI：
10.1016/j.neuro.2006.03.010
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发表时间：
2006-07-01
期刊：
影响因子：
3.4
通讯作者：
Ohta, Shigeru
中科院分区：
文献类型：
--
作者：
Nakatsu, Yusuke;Kotake, Yaichiro;Ohta, Shigeru
三丁基锡是一种干扰内分泌的化学品，已被用作热稳定剂、农业杀虫剂和油漆的成分。在这项研究中，我们调查钙蛋白酶是否参与三丁基锡毒性未分化的PC 12细胞。三丁基锡（2 μ M）诱导增加乳酸脱氢酶释放，细胞毒性的标志物，在PC 12细胞中以时间依赖性的方式。它还诱导钙蛋白酶激活的剂量依赖性的方式，和钙蛋白酶抑制剂，MDL 28170（40 μ M），降低细胞毒性，表明钙蛋白酶参与三丁基锡在PC 12细胞中的毒性。由于钙蛋白酶是一种钙依赖性蛋白酶，我们研究了EGTA（一种细胞外钙离子螯合剂）和BAPTA-AM（一种细胞内钙离子螯合剂）的作用。BAPTA-AM（50 μ M），但不是由EGTA（1 mM），三丁基锡诱导的钙蛋白酶激活减少，这表明钙蛋白酶激活与细胞内Ca 2+储存的钙释放。此外，我们还研究了caspase-3和calpain之间的关系。抑制caspase-3可降低三丁基锡诱导的钙蛋白酶活性。总之，我们已经证明，三丁基锡通过钙蛋白酶激活诱导细胞死亡，和细胞内Ca 2+的增加和caspase-3激活所需的钙蛋白酶激活三丁基锡。(c)2006年爱思唯尔公司All rights reserved.
Tributyltin, an endocrine-disrupting chemical, has been used as a heat stabilizer, agricultural pesticide and component of antifouling paints. In this study, we investigated whether calpain is involved in tributyltin toxicity in undifferentiated PC12 cells. Tributyltin (2 mu M) induced an increase of lactate dehydrogenase release, a marker of cytotoxicity, in PC 12 cells in a time-dependent manner. It also induced calpain activation in a dose-dependent manner, and a calpain inhibitor, MDL28170 (40 mu M), decreased the cellular toxicity, suggesting that calpain is involved in tributyltin toxicity in PC12 cells. Because calpain is a calcium-dependent protease, we examined the effect of EGTA, an extracellular Ca2+ chelator and BAPTA-AM, an intracellular Ca2+ chelator. Calpain activation induced by tributyltin was decreased by BAPTA-AM (50 mu M), but not by EGTA (1 mM), suggesting that calpain activation is associated with calcium release from intracellular Ca2+ stores. Further, we investigated the relationship between caspase-3 and calpain. Inhibition of caspase-3 reduced calpain activity induced by tributyltin. In conclusion, we have demonstrated that tributyltin induced cell death through calpain activation, and that intracellular Ca2+ increase and caspase-3 activation are required for calpain activation by tributyltin. (c) 2006 Elsevier Inc. All rights reserved.