TY - JOUR
T1 - Preconditioning of endoplasmic reticulum stress protects against acrylonitrile-induced cytotoxicity in primary rat astrocytes
T2 - The role of autophagy
AU - Yu, Bai
AU - Wenjun, Zhao
AU - Changsheng, Yin
AU - Yuntao, Fang
AU - Jing, Ma
AU - Ben, Li
AU - Hai, Qian
AU - Guangwei, Xing
AU - Suhua, Wang
AU - Fang, Li
AU - Aschner, Michael
AU - Rongzhu, Lu
N1 - Funding Information:
We are grateful to Dr. Bertrand Mollereau and Dr. Peter Spencer for editorial assistance. This work was supported in part by the Natural Science Foundation of China (No. 30872139 , No. 81273124 , No. 81302459 ) and the National Institute of Environmental Health Sciences ( ES10563 and ES07331 ), the China Postdoctoral Science Foundation ( 2013M540513 , 2015T80523 ), and the Science Foundation for Postdoctors in Jiangsu Province ( 1302151C ), Research Foundation for Advanced Talents in Jiangsu University ( 13JDG024 ).
Publisher Copyright:
© 2016 Elsevier B.V.
PY - 2016/7/1
Y1 - 2016/7/1
N2 - This study explored the protective effects of endoplasmic reticulum (ER) stress preconditioning induced by 2-deoxy-. d-glucose (2-DG) or oxidized dithiothreitol (DTTox) on acrylonitrile (AN)-induced cytotocity in primary rat astrocytes. Cells were pretreated with 2-DG or DTTox for different times at various concentration. Next, astrocytes were treated with 2.5 mM AN for an additional 12 h. Cell viability and cytotoxicity were assessed by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) reduction and lactate dehydrogenase (LDH) leakage, respectively. Reactive oxygen species (ROS) and mitochondrial membrane potential (δΨm) were determined. Expression of glucose-regulated protein 78 (GRP78), phosphorylated-eukaryotic translation initiation factor 2α (p-eIF2α), microtubule-associated protein light chain 3 (LC3), P62, and Beclin1 were used to assess autophagy. In addition, 3-methyadenine (3-MA), an autophagy-specific inhibitor, was used to assess the role of autophagy in ER stress preconditioning-induced protection against AN cytotoxicity. The results showed that AN alone significantly decreased astrocytic viability and enhanced cytotoxicity. Compared to the AN-alone group, preconditioning with 2-DG or DTTox significantly increased cell viability and reduced cytotoxicity to indistinguishable levels. Decreased ROS generation and increased δΨm were also inherent to ER stress preconditioning with these compounds. Furthermore, autophagy was activated by both 2-DG and DTTox. Blockage of autophagy attenuated the protection afforded by 2-DG or DTTox preconditioning in AN-treated astrocytes. These results establish that ER stress preconditioning affords cellular protection against AN, and that activation of autophagy mediates the cytoprotection. Modulation of ER stress and resultant activation of autophagy may be a novel target for to ameliorate AN toxicity.
AB - This study explored the protective effects of endoplasmic reticulum (ER) stress preconditioning induced by 2-deoxy-. d-glucose (2-DG) or oxidized dithiothreitol (DTTox) on acrylonitrile (AN)-induced cytotocity in primary rat astrocytes. Cells were pretreated with 2-DG or DTTox for different times at various concentration. Next, astrocytes were treated with 2.5 mM AN for an additional 12 h. Cell viability and cytotoxicity were assessed by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) reduction and lactate dehydrogenase (LDH) leakage, respectively. Reactive oxygen species (ROS) and mitochondrial membrane potential (δΨm) were determined. Expression of glucose-regulated protein 78 (GRP78), phosphorylated-eukaryotic translation initiation factor 2α (p-eIF2α), microtubule-associated protein light chain 3 (LC3), P62, and Beclin1 were used to assess autophagy. In addition, 3-methyadenine (3-MA), an autophagy-specific inhibitor, was used to assess the role of autophagy in ER stress preconditioning-induced protection against AN cytotoxicity. The results showed that AN alone significantly decreased astrocytic viability and enhanced cytotoxicity. Compared to the AN-alone group, preconditioning with 2-DG or DTTox significantly increased cell viability and reduced cytotoxicity to indistinguishable levels. Decreased ROS generation and increased δΨm were also inherent to ER stress preconditioning with these compounds. Furthermore, autophagy was activated by both 2-DG and DTTox. Blockage of autophagy attenuated the protection afforded by 2-DG or DTTox preconditioning in AN-treated astrocytes. These results establish that ER stress preconditioning affords cellular protection against AN, and that activation of autophagy mediates the cytoprotection. Modulation of ER stress and resultant activation of autophagy may be a novel target for to ameliorate AN toxicity.
KW - Acrylonitrile
KW - Autophagy
KW - Endoplasmic reticulum stress
KW - Preconditioning
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U2 - 10.1016/j.neuro.2016.05.020
DO - 10.1016/j.neuro.2016.05.020
M3 - Article
C2 - 27260289
AN - SCOPUS:84973562840
SN - 0161-813X
VL - 55
SP - 112
EP - 121
JO - NeuroToxicology
JF - NeuroToxicology
ER -