Akt/GSK3β signaling is involved in fipronil-induced apoptotic cell death of human neuroblastoma SH-SY5Y cells
Akt/GSK3β signaling is involved in fipronil-induced apoptotic cell death of human neuroblastoma SH-SY5Y cells
复制标题
DOI：
10.1016/j.toxlet.2011.01.030
复制
发表时间：
2011-04-25
影响因子：
3.5
通讯作者：
Koh, Hyun Chul
中科院分区：
文献类型：
--
作者：
Lee, Jeong Eun;Kang, Jin Sun;Koh, Hyun Chul
氟虫腈（FPN）是一种作用于昆虫γ-氨基丁酸（GABA）受体的苯基吡唑类杀虫剂。虽然FPN的作用仅限于昆虫神经或肌肉递质系统，但少数研究评估了这种神经毒物对神经细胞死亡的影响。为了确定FPN诱导的神经元细胞死亡的机制，我们研究了活性氧（ROS）是否在FPN诱导的细胞凋亡中发挥作用，使用人多巴胺能SH-SY 5 Y细胞的体外模型。FPN对这些细胞具有细胞毒性，且呈浓度依赖性。此外，FPN处理显著降低酪氨酸羟化酶（TH）的表达，而谷氨酸脱羧酶65（GAD 65）的表达没有变化。FPN诱导的多巴胺能细胞死亡与ROS产生增加有关，因为用N-乙酰半胱氨酸（NAC）预处理，抗氧化剂，减少细胞死亡。FPN处理后，细胞和培养液中多巴胺（DA）水平均显著降低，NAC预处理可阻断DA的氧化作用。我们发现，细胞死亡响应FPN是由于细胞凋亡，因为FPN增加细胞色素C释放到胞质溶胶和激活caspase-3。同时，还可引起p53蛋白在细胞核内的聚集，并使Bcl-2蛋白水平呈浓度依赖性降低。此外，FPN改变Akt/糖原合成酶激酶-3（GSK 3 β）磷酸化水平。FPN降低了Ser 473上Akt的磷酸化，并且与Akt的失活平行，在用FPN处理后，使GSK 3 β失活的Ser 9上的GSK 3 β的磷酸化降低。此外，GSK 3 β信号的抑制保护细胞免于FPN诱导的细胞死亡。这些结果表明，GSK 3 β活性的调节可以控制由FPN诱导的氧化应激诱导的细胞凋亡，与神经元细胞死亡相关。皇冠版权所有（C）2011由爱思唯尔爱尔兰有限公司出版。保留所有权利。
Fipronil (FPN) is a phenylpyrazole insecticide acted on insect gamma-aminobutyric acid (GABA) receptors. Although action of FPN is restricted on insect neuronal or muscular transmitter system, a few studies have assessed the effects of this neurotoxicant on neuronal cell death. To determine the mechanisms underlying FPN-induced neuronal cell death, we investigated whether reactive oxygen species (ROS) plays a role in FPN-induced apoptosis, using an in vitro model of human dopaminergic SH-SY5Y cells. FPN was cytotoxic to these cells and its cytotoxicity showed a concentration-dependent manner. Additionally, FPN treatment significantly decreased the tyrosine hydroxylase (TH) expression without change of glutamic acid decarboxylase 65 (GAD65) expression. FPN-induced dopaminergic cell death involved in increase of ROS generation since pretreatment with N-acetyl cysteine (NAC), an anti-oxidant, reduced cell death. After FPN treatment, dopamine (DA) levels decreased significantly in both cell and culture media, and oxidative effects of DA were blocked by NAC pretreatment. We showed that cell death in response to FPN was due to apoptosis since FPN increased cytochrome c release into the cytosol and activated caspase-3. It also led to nuclear accumulation of p53 and reduced the level of Bcl-2 protein in a concentration-dependent manner. Additionally, FPN altered the level of Akt/glycogen synthase kinase-3 (GSK3 beta) phosphorylation. FPN reduced the Akt phosphorylation on Ser473, and in parallel with the inactivation of Akt, phosphorylation of GSK3 beta on Ser9 which inactivates GSK3 beta, decreased after treatment with FPN. Furthermore, inhibition of the GSK3 beta signal protected the cell against FPN-induced cell death. These results suggest that regulation of GSK3 beta activity may control the apoptosis induced by FPN-induced oxidative stress associated with neuronal cell death. Crown Copyright (C) 2011 Published by Elsevier Ireland Ltd. All rights reserved.