Quercetin inhibition of ROS-dependent and -independent apoptosis in rat glioma C6 cells

Tong Jong Chen, Jing Yueh Jeng, Cheng Wei Lin, Chin Yen Wu, Yen Chou Chen

Research output: Contribution to journalArticle

137 Citations (Scopus)

Abstract

In the present study, we investigated the protective mechanism of quercetin (QUE) and its glycosides, rutin (RUT) and quercitrin (QUI), on reactive oxygen species (ROS)-dependent (H 2O 2) and -independent (chemical anoxia) cell death in rat glioma C6 cells. Induction of HO-1 protein expression was detected in QUE- but not RUT- or QUI-treated C6 cells, and this was prevented by cycloheximide and actinomycin D. Incubation of C6 cells with QUE, but not RUT or QUI, protected C6 cells from H 2O 2- and chemical anoxia-induced cytotoxicity according to the MTT and LDH release assays. Apoptotic characteristics including chromatin condensation, DNA ladders, and hypodiploid cells appeared in H 2O 2-and chemical anoxia-treated C6 cells, and those events were significantly suppressed by adding QUE (but not RUT or QUI). Increases in caspase 3, 8, and 9 enzyme activities with decreases in pro-PARP and pro-caspase 3 protein levels and an increase in cleaved D4-GDI protein were identified in H 2O 2-and chemical anoxia-treated C6 cells, and these were blocked by the addition of QUE, but not by RUT or QUI. Intracellular peroxide levels increased with H 2O 2 and decreased with chemical anoxia, and the addition of QUE reduced the intracellular peroxide levels induced by H 2O 2. Results of an anti-DPPH radical assay showed that QUE, RUT, and QUI dose-dependently inhibited the production of DPPH radicals in vitro; however, QUE (but not RUT or QUI) prevention of DNA damage induced by OH radicals was identified with a plasmid digestion assay. Increases in phosphorylated ERK and p53 protein expressions were detected in H 2O 2- but not chemical anoxia-treated C6 cells, and the addition of QUE significantly blocked H 2O 2-induced phosphorylated ERK and p53 protein expressions. Adding the HO-1 inhibitors, SnPP, CoPP, and ZnPP, reversed the protective effect of QUE against H 2O 2- and chemical anoxia-induced cell death according to the MTT assay and morphological observations. Additionally, QUE exhibited inhibitory effects on LPS/TPA-induced transformation in accordance with a decrease in MMP-9 enzyme activity and iNOS protein expression in C6 cells. Taken together, the results of this study suggest that QUE exhibits an inhibitory effect on both ROS-dependent and -independent cell death, and induction of HO-1 protein expression is involved.

Original languageEnglish
Pages (from-to)113-126
Number of pages14
JournalToxicology
Volume223
Issue number1-2
DOIs
Publication statusPublished - Jun 1 2006

Fingerprint

Quercetin
Glioma
Rats
Reactive Oxygen Species
Rutin
Apoptosis
Assays
Cell death
Proteins
Cell Death
Peroxides
Enzyme activity
Caspase 3
Guanine Nucleotide Dissociation Inhibitors
Caspase 9
Caspase 8
DNA
Ladders
Dactinomycin
Enzymes

Keywords

  • Apoptosis
  • Chemical anoxia
  • HO-1
  • Quercetin
  • ROS

ASJC Scopus subject areas

  • Toxicology

Cite this

Quercetin inhibition of ROS-dependent and -independent apoptosis in rat glioma C6 cells. / Chen, Tong Jong; Jeng, Jing Yueh; Lin, Cheng Wei; Wu, Chin Yen; Chen, Yen Chou.

In: Toxicology, Vol. 223, No. 1-2, 01.06.2006, p. 113-126.

Research output: Contribution to journalArticle

Chen, Tong Jong ; Jeng, Jing Yueh ; Lin, Cheng Wei ; Wu, Chin Yen ; Chen, Yen Chou. / Quercetin inhibition of ROS-dependent and -independent apoptosis in rat glioma C6 cells. In: Toxicology. 2006 ; Vol. 223, No. 1-2. pp. 113-126.
@article{783b7c83f89341e1b8e64b6b0eae781a,
title = "Quercetin inhibition of ROS-dependent and -independent apoptosis in rat glioma C6 cells",
abstract = "In the present study, we investigated the protective mechanism of quercetin (QUE) and its glycosides, rutin (RUT) and quercitrin (QUI), on reactive oxygen species (ROS)-dependent (H 2O 2) and -independent (chemical anoxia) cell death in rat glioma C6 cells. Induction of HO-1 protein expression was detected in QUE- but not RUT- or QUI-treated C6 cells, and this was prevented by cycloheximide and actinomycin D. Incubation of C6 cells with QUE, but not RUT or QUI, protected C6 cells from H 2O 2- and chemical anoxia-induced cytotoxicity according to the MTT and LDH release assays. Apoptotic characteristics including chromatin condensation, DNA ladders, and hypodiploid cells appeared in H 2O 2-and chemical anoxia-treated C6 cells, and those events were significantly suppressed by adding QUE (but not RUT or QUI). Increases in caspase 3, 8, and 9 enzyme activities with decreases in pro-PARP and pro-caspase 3 protein levels and an increase in cleaved D4-GDI protein were identified in H 2O 2-and chemical anoxia-treated C6 cells, and these were blocked by the addition of QUE, but not by RUT or QUI. Intracellular peroxide levels increased with H 2O 2 and decreased with chemical anoxia, and the addition of QUE reduced the intracellular peroxide levels induced by H 2O 2. Results of an anti-DPPH radical assay showed that QUE, RUT, and QUI dose-dependently inhibited the production of DPPH radicals in vitro; however, QUE (but not RUT or QUI) prevention of DNA damage induced by OH radicals was identified with a plasmid digestion assay. Increases in phosphorylated ERK and p53 protein expressions were detected in H 2O 2- but not chemical anoxia-treated C6 cells, and the addition of QUE significantly blocked H 2O 2-induced phosphorylated ERK and p53 protein expressions. Adding the HO-1 inhibitors, SnPP, CoPP, and ZnPP, reversed the protective effect of QUE against H 2O 2- and chemical anoxia-induced cell death according to the MTT assay and morphological observations. Additionally, QUE exhibited inhibitory effects on LPS/TPA-induced transformation in accordance with a decrease in MMP-9 enzyme activity and iNOS protein expression in C6 cells. Taken together, the results of this study suggest that QUE exhibits an inhibitory effect on both ROS-dependent and -independent cell death, and induction of HO-1 protein expression is involved.",
keywords = "Apoptosis, Chemical anoxia, HO-1, Quercetin, ROS",
author = "Chen, {Tong Jong} and Jeng, {Jing Yueh} and Lin, {Cheng Wei} and Wu, {Chin Yen} and Chen, {Yen Chou}",
year = "2006",
month = "6",
day = "1",
doi = "10.1016/j.tox.2006.03.007",
language = "English",
volume = "223",
pages = "113--126",
journal = "Toxicology",
issn = "0300-483X",
publisher = "Elsevier Ireland Ltd",
number = "1-2",

}

TY - JOUR

T1 - Quercetin inhibition of ROS-dependent and -independent apoptosis in rat glioma C6 cells

AU - Chen, Tong Jong

AU - Jeng, Jing Yueh

AU - Lin, Cheng Wei

AU - Wu, Chin Yen

AU - Chen, Yen Chou

PY - 2006/6/1

Y1 - 2006/6/1

N2 - In the present study, we investigated the protective mechanism of quercetin (QUE) and its glycosides, rutin (RUT) and quercitrin (QUI), on reactive oxygen species (ROS)-dependent (H 2O 2) and -independent (chemical anoxia) cell death in rat glioma C6 cells. Induction of HO-1 protein expression was detected in QUE- but not RUT- or QUI-treated C6 cells, and this was prevented by cycloheximide and actinomycin D. Incubation of C6 cells with QUE, but not RUT or QUI, protected C6 cells from H 2O 2- and chemical anoxia-induced cytotoxicity according to the MTT and LDH release assays. Apoptotic characteristics including chromatin condensation, DNA ladders, and hypodiploid cells appeared in H 2O 2-and chemical anoxia-treated C6 cells, and those events were significantly suppressed by adding QUE (but not RUT or QUI). Increases in caspase 3, 8, and 9 enzyme activities with decreases in pro-PARP and pro-caspase 3 protein levels and an increase in cleaved D4-GDI protein were identified in H 2O 2-and chemical anoxia-treated C6 cells, and these were blocked by the addition of QUE, but not by RUT or QUI. Intracellular peroxide levels increased with H 2O 2 and decreased with chemical anoxia, and the addition of QUE reduced the intracellular peroxide levels induced by H 2O 2. Results of an anti-DPPH radical assay showed that QUE, RUT, and QUI dose-dependently inhibited the production of DPPH radicals in vitro; however, QUE (but not RUT or QUI) prevention of DNA damage induced by OH radicals was identified with a plasmid digestion assay. Increases in phosphorylated ERK and p53 protein expressions were detected in H 2O 2- but not chemical anoxia-treated C6 cells, and the addition of QUE significantly blocked H 2O 2-induced phosphorylated ERK and p53 protein expressions. Adding the HO-1 inhibitors, SnPP, CoPP, and ZnPP, reversed the protective effect of QUE against H 2O 2- and chemical anoxia-induced cell death according to the MTT assay and morphological observations. Additionally, QUE exhibited inhibitory effects on LPS/TPA-induced transformation in accordance with a decrease in MMP-9 enzyme activity and iNOS protein expression in C6 cells. Taken together, the results of this study suggest that QUE exhibits an inhibitory effect on both ROS-dependent and -independent cell death, and induction of HO-1 protein expression is involved.

AB - In the present study, we investigated the protective mechanism of quercetin (QUE) and its glycosides, rutin (RUT) and quercitrin (QUI), on reactive oxygen species (ROS)-dependent (H 2O 2) and -independent (chemical anoxia) cell death in rat glioma C6 cells. Induction of HO-1 protein expression was detected in QUE- but not RUT- or QUI-treated C6 cells, and this was prevented by cycloheximide and actinomycin D. Incubation of C6 cells with QUE, but not RUT or QUI, protected C6 cells from H 2O 2- and chemical anoxia-induced cytotoxicity according to the MTT and LDH release assays. Apoptotic characteristics including chromatin condensation, DNA ladders, and hypodiploid cells appeared in H 2O 2-and chemical anoxia-treated C6 cells, and those events were significantly suppressed by adding QUE (but not RUT or QUI). Increases in caspase 3, 8, and 9 enzyme activities with decreases in pro-PARP and pro-caspase 3 protein levels and an increase in cleaved D4-GDI protein were identified in H 2O 2-and chemical anoxia-treated C6 cells, and these were blocked by the addition of QUE, but not by RUT or QUI. Intracellular peroxide levels increased with H 2O 2 and decreased with chemical anoxia, and the addition of QUE reduced the intracellular peroxide levels induced by H 2O 2. Results of an anti-DPPH radical assay showed that QUE, RUT, and QUI dose-dependently inhibited the production of DPPH radicals in vitro; however, QUE (but not RUT or QUI) prevention of DNA damage induced by OH radicals was identified with a plasmid digestion assay. Increases in phosphorylated ERK and p53 protein expressions were detected in H 2O 2- but not chemical anoxia-treated C6 cells, and the addition of QUE significantly blocked H 2O 2-induced phosphorylated ERK and p53 protein expressions. Adding the HO-1 inhibitors, SnPP, CoPP, and ZnPP, reversed the protective effect of QUE against H 2O 2- and chemical anoxia-induced cell death according to the MTT assay and morphological observations. Additionally, QUE exhibited inhibitory effects on LPS/TPA-induced transformation in accordance with a decrease in MMP-9 enzyme activity and iNOS protein expression in C6 cells. Taken together, the results of this study suggest that QUE exhibits an inhibitory effect on both ROS-dependent and -independent cell death, and induction of HO-1 protein expression is involved.

KW - Apoptosis

KW - Chemical anoxia

KW - HO-1

KW - Quercetin

KW - ROS

UR - http://www.scopus.com/inward/record.url?scp=33646578517&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=33646578517&partnerID=8YFLogxK

U2 - 10.1016/j.tox.2006.03.007

DO - 10.1016/j.tox.2006.03.007

M3 - Article

C2 - 16647178

AN - SCOPUS:33646578517

VL - 223

SP - 113

EP - 126

JO - Toxicology

JF - Toxicology

SN - 0300-483X

IS - 1-2

ER -