Oxidative stress is involved in Dasatinib-induced apoptosis in rat primary hepatocytes
Oxidative stress is involved in Dasatinib-induced apoptosis in rat primary hepatocytes
复制标题
氧化应激参与达沙替尼诱导的大鼠原代肝细胞凋亡
DOI：
10.1016/j.taap.2012.04.010
复制
发表时间：
2012-06-15
影响因子：
3.8
通讯作者：
He, Qiaojun
中科院分区：
文献类型：
--
作者：
Xue, Tao;Luo, Peihua;He, Qiaojun
达沙替尼是一种BCR-ABL和SRC激酶的多靶点抑制剂，具有抗肿瘤活性，可延长慢性粒细胞白血病（CML）和费城染色体阳性急性淋巴细胞白血病（ALL）患者的生存期。然而，一些患者患有肝毒性，其通过未知的机制发生。在本研究中，我们发现达沙替尼在体外和体内都可以诱导肝毒性。达沙替尼降低大鼠原代肝细胞的细胞活力，在体外诱导丙氨酸氨基转移酶（ALT）和乳酸脱氢酶（LDH）的释放，并在体内触发Sprague-Dawley大鼠肝细胞的气球样变性。检测凋亡标志物（染色质凝聚、裂解的caspase-3和裂解的PARP），以表明达沙替尼在体外肝细胞中诱导的损伤是由凋亡介导的。使用末端脱氧核苷酸转移酶介导的dUTP缺口末端标记（TUNEL）测定在体内进一步验证了该结果。我们发现达沙替尼可显著增加肝细胞内活性氧（ROS）水平，降低细胞内谷胱甘肽（GSH）含量，减弱超氧化物歧化酶（SOD）活性，产生脂质过氧化产物丙二醛（MDA），降低线粒体膜电位，以及与氧化应激和存活相关的活化核因子红细胞2相关因子2（Nrf 2）和促分裂原活化蛋白激酶（MAPK）。这些结果证实，氧化应激在达沙替尼介导的肝毒性中起关键作用。N-乙酰半胱氨酸（NAC）是一种典型的抗氧化剂，可以清除自由基，减轻氧化应激，保护肝细胞免受达沙替尼诱导的损伤。因此，缓解氧化应激是降低达沙替尼诱导的肝毒性的可行策略。皇冠版权所有（C）2012由Elsevier Inc.发布。All rights reserved.
Dasatinib, a multitargeted inhibitor of BCR-ABL and SRC kinases, exhibits antitumor activity and extends the survival of patients with chronic myeloid leukemia (CML) and Philadelphia chromosome-positive acute lymphoblastic leukemia (ALL). However, some patients suffer from hepatotoxicity, which occurs through an unknown mechanism. In the present study, we found that Dasatinib could induce hepatotoxicity both in vitro and in vivo. Dasatinib reduced the cell viability of rat primary hepatocytes, induced the release of alanine aminotransferase (ALT) and lactate dehydrogenase (LDH) in vitro, and triggered the ballooning degeneration of hepatocytes in Sprague-Dawley rats in vivo. Apoptotic markers (chromatin condensation, cleaved caspase-3 and cleaved PARP) were detected to indicate that the injury induced by Dasatinib in hepatocytes in vitro was mediated by apoptosis. This result was further validated in vivo using terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling (TUNEL) assays. Here we found that Dasatinib dramatically increased the level of reactive oxygen species (ROS) in hepatocytes, reduced the intracellular glutathione (GSH) content, attenuated the activity of superoxide dismutase (SOD), generated malondialdehyde (MDA), a product of lipid peroxidation, decreased the mitochondrial membrane potential, and activated nuclear factor erythroid 2-related factor 2 (Nrf2) and mitogen-activated protein kinases (MAPK) related to oxidative stress and survival. These results confirm that oxidative stress plays a pivotal role in Dasatinib-mediated hepatotoxicity. N-acetylcysteine (NAC), a typical antioxidant, can scavenge free radicals, attenuate oxidative stress, and protect hepatocytes against Dasatinib-induced injury. Thus, relieving oxidative stress is a viable strategy for reducing Dasatinib-induced hepatotoxicity. Crown Copyright (C) 2012 Published by Elsevier Inc. All rights reserved.