Enhanced toxicity and ROS generation by doxorubicin in primary cultures of cardiomyocytes from neonatal metallothionein-I/II null mice
Enhanced toxicity and ROS generation by doxorubicin in primary cultures of cardiomyocytes from neonatal metallothionein-I/II null mice
复制标题
DOI：
10.1016/j.tiv.2010.06.009
复制
发表时间：
2010-09-01
影响因子：
3.2
通讯作者：
Peng, Shuangqing
中科院分区：
文献类型：
--
作者：
Fu, Ze;Guo, Jiabin;Peng, Shuangqing
多柔比星（Dox）是一种有效的抗癌药物，但其剂量依赖性心脏毒性限制了其临床应用。我们以前发现，金属硫蛋白-I/II（MT-I/II）空小鼠更容易受到Dox诱导的心肌病，但它是未知的，是否耗尽MT会敏感心肌细胞对Dox毒性在体外，因为MT的保护作用仍然存在争议。在本研究中，从新生的MT-I/II null（MT-/-）和相应的野生型（MT+/+）小鼠的心肌细胞的原代培养系统的建立，以明确地确定MT缺乏对Dox诱导的毒性的影响。MT-/-心肌细胞中的MT浓度比MT+/+心肌细胞中的MT浓度低约2.5倍。MT-/-心肌细胞比MT+/+心肌细胞对Dox诱导的细胞毒性更敏感，通过形态学改变、乳酸脱氢酶渗漏、细胞活力和凋亡来测量。Dox时间和浓度依赖性地增加MT+/+心肌细胞中活性氧（ROS）的形成，并且这种作用在MT-/-心肌细胞中被夸大。抗氧化剂N-乙酰半胱氨酸（NAC）和谷胱甘肽（GSH）可显著挽救MT+/+心肌细胞免于Dox诱导的细胞死亡和ROS产生，但MT（-/-）心肌细胞则无此作用。这些结果表明，基础MT对Dox诱导的心肌细胞毒性具有保护作用，特别强调了MT作为细胞抗氧化剂在清除ROS方面的重要作用。(C)2010爱思唯尔有限公司保留所有权利。
The clinical use of doxorubicin (Dox), a potent anticancer drug, is limited by its concurrent dose-dependent cardiotoxicity. We previously found that metallothionein-I/II (MT-I/II) null mice are more vulnerable to Dox-induced cardiomyopathy, but it is unknown whether depletion of MT would sensitize cardiomyocytes to Dox toxicity in vitro since the protective effect of MT still remains controversial. In the present study, a primary culture system of cardiomyocytes from neonatal MT-I/II null (MT-/-) and corresponding wild type (MT+/+) mice was established to unequivocally determine the effect of MT deficiency on Dox-induced toxicity. MT concentrations in the MT-/- cardiomyocytes were about 2.5-fold lower than those in MT+/+ cardiomyocytes. MT-/- cardiomyocytes were more sensitive to Dox-induced cytotoxicity than MT+/+ cardiomyocytes as measured by morphological alterations, lactate dehydrogenase leakage, cell viability, and apoptosis. Dox time- and concentration-dependently increased reactive oxygen species (ROS) formation in MT+/+ cardiomyocytes, and this effect was exaggerated in MT-/- cardiomyocytes. Antioxidant N-acetylcysteine (NAC) and glutathione (GSH) significantly rescued MT+/+ but not MT(-/-)cardiomyocytes from Dox-induced cell death and ROS generation. These findings suggest that basal MT provide protection against Dox-induced toxicity in cardiomyocytes, particularly highlight the important role of MT as a cellular antioxidant on scavenging ROS. (C) 2010 Elsevier Ltd. All rights reserved.