Differential protection of pre- versus post-treatment with curcumin, Trolox, and N-acetylcysteine against acrylonitrile-induced cytotoxicity in primary rat astrocytes

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

Research output: Contribution to journalArticle

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Abstract

Objective: This study was designed to examine the differential protection of pre- versus post-treatment with three different antioxidants, curcumin (CUR), Trolox, and N-acetylcysteine (NAC), on acrylonitrile (AN)-induced cytotoxicity in primary rat astrocytes. Methods: Primary astrocyte cultures were treated with CUR, Trolox and NAC for 4 h prior to, or following 24 h treatment with AN (2.5 mM). 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT), lactate dehydrogenase (LDH) release, lipid peroxidation, glutathione, reactive oxygen species (ROS) and mitochondrial membrane potential were measured to evaluate protection associated with the three antioxidants. Knockdown of Nrf2 expression by liposome transfection with siRNA was used to confirm the role of Nrf2 activation in the protection associated with the three antioxidants. Results: Compared with AN treatment alone, pre-treatment with CUR at either concentration significantly increased cell viability and mitochondrial membrane potential, and reduced glutathione levels; lipid peroxidation and ROS production were significantly decreased as well. NAC also showed significant efficacy in attenuating AN-induced toxicity at higher concentration. However, pre-treatment with Trolox failed to ameliorate the AN-induced toxicity. When post-treatment with Trolox, this antioxidant led to significant protective effects at both concentrations, while CUR and NAC were efficacious only at the higher concentrations. Knockdown of Nrf2 only abolished the protective effects of CUR pre-treatment on AN-induced cytotoxicity, while the protective effects of NAC and Trolox pre-treatment groups showed no differences between the Nrf2-knockdown and non-knockdown treatments. Conclusions: The selected antioxidants exert differential cellular protection when administered prior or subsequent to AN-induced cytotoxic events in decreasing cellular viability, antioxidative capacity and mitochondrial function, enhanced cytotoxicity and ROS production. These results suggest that antioxidants should be carefully chosen for their efficacy in preventing or diminishing oxidative damage caused by AN. The differential effect of pre- and post-treatment may be attributed to activation of the Nrf2 signaling pathway.

Original languageEnglish (US)
Pages (from-to)58-66
Number of pages9
JournalNeuroToxicology
Volume51
DOIs
StatePublished - Dec 1 2015

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Acrylonitrile
Curcumin
Acetylcysteine
Cytotoxicity
Astrocytes
Rats
Antioxidants
Reactive Oxygen Species
Therapeutics
Glutathione
Toxicity
Mitochondrial Membrane Potential
Chemical activation
Membranes
Lipid Peroxidation
Lipids
6-hydroxy-2,5,7,8-tetramethylchroman-2-carboxylic acid
L-Lactate Dehydrogenase
Liposomes
Small Interfering RNA

Keywords

  • Acrylonitrile
  • Curcumin
  • N-Acetylcysteine
  • Post-treatment
  • Pre-treatment
  • Trolox

ASJC Scopus subject areas

  • Neuroscience(all)
  • Toxicology

Cite this

Differential protection of pre- versus post-treatment with curcumin, Trolox, and N-acetylcysteine against acrylonitrile-induced cytotoxicity in primary rat astrocytes. / Yu, Bai; Changsheng, Yin; Wenjun, Zhao; Ben, Li; Hai, Qian; Jing, Ma; Guangwei, Xing; Shuhua, Wang; Fang, Li; Aschner, Michael; Rongzhu, Lu.

In: NeuroToxicology, Vol. 51, 01.12.2015, p. 58-66.

Research output: Contribution to journalArticle

Yu, Bai ; Changsheng, Yin ; Wenjun, Zhao ; Ben, Li ; Hai, Qian ; Jing, Ma ; Guangwei, Xing ; Shuhua, Wang ; Fang, Li ; Aschner, Michael ; Rongzhu, Lu. / Differential protection of pre- versus post-treatment with curcumin, Trolox, and N-acetylcysteine against acrylonitrile-induced cytotoxicity in primary rat astrocytes. In: NeuroToxicology. 2015 ; Vol. 51. pp. 58-66.
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T1 - Differential protection of pre- versus post-treatment with curcumin, Trolox, and N-acetylcysteine against acrylonitrile-induced cytotoxicity in primary rat astrocytes

AU - Yu, Bai

AU - Changsheng, Yin

AU - Wenjun, Zhao

AU - Ben, Li

AU - Hai, Qian

AU - Jing, Ma

AU - Guangwei, Xing

AU - Shuhua, Wang

AU - Fang, Li

AU - Aschner, Michael

AU - Rongzhu, Lu

PY - 2015/12/1

Y1 - 2015/12/1

N2 - Objective: This study was designed to examine the differential protection of pre- versus post-treatment with three different antioxidants, curcumin (CUR), Trolox, and N-acetylcysteine (NAC), on acrylonitrile (AN)-induced cytotoxicity in primary rat astrocytes. Methods: Primary astrocyte cultures were treated with CUR, Trolox and NAC for 4 h prior to, or following 24 h treatment with AN (2.5 mM). 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT), lactate dehydrogenase (LDH) release, lipid peroxidation, glutathione, reactive oxygen species (ROS) and mitochondrial membrane potential were measured to evaluate protection associated with the three antioxidants. Knockdown of Nrf2 expression by liposome transfection with siRNA was used to confirm the role of Nrf2 activation in the protection associated with the three antioxidants. Results: Compared with AN treatment alone, pre-treatment with CUR at either concentration significantly increased cell viability and mitochondrial membrane potential, and reduced glutathione levels; lipid peroxidation and ROS production were significantly decreased as well. NAC also showed significant efficacy in attenuating AN-induced toxicity at higher concentration. However, pre-treatment with Trolox failed to ameliorate the AN-induced toxicity. When post-treatment with Trolox, this antioxidant led to significant protective effects at both concentrations, while CUR and NAC were efficacious only at the higher concentrations. Knockdown of Nrf2 only abolished the protective effects of CUR pre-treatment on AN-induced cytotoxicity, while the protective effects of NAC and Trolox pre-treatment groups showed no differences between the Nrf2-knockdown and non-knockdown treatments. Conclusions: The selected antioxidants exert differential cellular protection when administered prior or subsequent to AN-induced cytotoxic events in decreasing cellular viability, antioxidative capacity and mitochondrial function, enhanced cytotoxicity and ROS production. These results suggest that antioxidants should be carefully chosen for their efficacy in preventing or diminishing oxidative damage caused by AN. The differential effect of pre- and post-treatment may be attributed to activation of the Nrf2 signaling pathway.

AB - Objective: This study was designed to examine the differential protection of pre- versus post-treatment with three different antioxidants, curcumin (CUR), Trolox, and N-acetylcysteine (NAC), on acrylonitrile (AN)-induced cytotoxicity in primary rat astrocytes. Methods: Primary astrocyte cultures were treated with CUR, Trolox and NAC for 4 h prior to, or following 24 h treatment with AN (2.5 mM). 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT), lactate dehydrogenase (LDH) release, lipid peroxidation, glutathione, reactive oxygen species (ROS) and mitochondrial membrane potential were measured to evaluate protection associated with the three antioxidants. Knockdown of Nrf2 expression by liposome transfection with siRNA was used to confirm the role of Nrf2 activation in the protection associated with the three antioxidants. Results: Compared with AN treatment alone, pre-treatment with CUR at either concentration significantly increased cell viability and mitochondrial membrane potential, and reduced glutathione levels; lipid peroxidation and ROS production were significantly decreased as well. NAC also showed significant efficacy in attenuating AN-induced toxicity at higher concentration. However, pre-treatment with Trolox failed to ameliorate the AN-induced toxicity. When post-treatment with Trolox, this antioxidant led to significant protective effects at both concentrations, while CUR and NAC were efficacious only at the higher concentrations. Knockdown of Nrf2 only abolished the protective effects of CUR pre-treatment on AN-induced cytotoxicity, while the protective effects of NAC and Trolox pre-treatment groups showed no differences between the Nrf2-knockdown and non-knockdown treatments. Conclusions: The selected antioxidants exert differential cellular protection when administered prior or subsequent to AN-induced cytotoxic events in decreasing cellular viability, antioxidative capacity and mitochondrial function, enhanced cytotoxicity and ROS production. These results suggest that antioxidants should be carefully chosen for their efficacy in preventing or diminishing oxidative damage caused by AN. The differential effect of pre- and post-treatment may be attributed to activation of the Nrf2 signaling pathway.

KW - Acrylonitrile

KW - Curcumin

KW - N-Acetylcysteine

KW - Post-treatment

KW - Pre-treatment

KW - Trolox

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