Preconditioning of endoplasmic reticulum stress protects against acrylonitrile-induced cytotoxicity in primary rat astrocytes

The role of autophagy

Bai Yu, Zhao Wenjun, Yin Changsheng, Fang Yuntao, Ma Jing, Li Ben, Qian Hai, Xing Guangwei, Wang Suhua, Li Fang, Michael Aschner, Lu Rongzhu

Research output: Contribution to journalArticle

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Abstract

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.

Original languageEnglish (US)
Pages (from-to)112-121
Number of pages10
JournalNeuroToxicology
Volume55
DOIs
StatePublished - Jul 1 2016

Fingerprint

Acrylonitrile
Endoplasmic Reticulum Stress
Autophagy
Cytotoxicity
Astrocytes
Rats
Reactive Oxygen Species
Cell Survival
Prokaryotic Initiation Factor-2
Chemical activation
Eukaryotic Initiation Factor-2
Cells
Eukaryotic Initiation Factors
Microtubule-Associated Proteins
Cytoprotection
Mitochondrial Membrane Potential
Dithiothreitol
L-Lactate Dehydrogenase
Toxicity
Modulation

Keywords

  • Acrylonitrile
  • Autophagy
  • Endoplasmic reticulum stress
  • Preconditioning

ASJC Scopus subject areas

  • Neuroscience(all)
  • Toxicology

Cite this

Preconditioning of endoplasmic reticulum stress protects against acrylonitrile-induced cytotoxicity in primary rat astrocytes : The role of autophagy. / Yu, Bai; Wenjun, Zhao; Changsheng, Yin; Yuntao, Fang; Jing, Ma; Ben, Li; Hai, Qian; Guangwei, Xing; Suhua, Wang; Fang, Li; Aschner, Michael; Rongzhu, Lu.

In: NeuroToxicology, Vol. 55, 01.07.2016, p. 112-121.

Research output: Contribution to journalArticle

Yu, B, Wenjun, Z, Changsheng, Y, Yuntao, F, Jing, M, Ben, L, Hai, Q, Guangwei, X, Suhua, W, Fang, L, Aschner, M & Rongzhu, L 2016, 'Preconditioning of endoplasmic reticulum stress protects against acrylonitrile-induced cytotoxicity in primary rat astrocytes: The role of autophagy', NeuroToxicology, vol. 55, pp. 112-121. https://doi.org/10.1016/j.neuro.2016.05.020
Yu, Bai ; Wenjun, Zhao ; Changsheng, Yin ; Yuntao, Fang ; Jing, Ma ; Ben, Li ; Hai, Qian ; Guangwei, Xing ; Suhua, Wang ; Fang, Li ; Aschner, Michael ; Rongzhu, Lu. / Preconditioning of endoplasmic reticulum stress protects against acrylonitrile-induced cytotoxicity in primary rat astrocytes : The role of autophagy. In: NeuroToxicology. 2016 ; Vol. 55. pp. 112-121.
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abstract = "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.",
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AU - Jing, Ma

AU - Ben, Li

AU - Hai, Qian

AU - Guangwei, Xing

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AU - Rongzhu, Lu

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