Intracellular pH-dependent peroxynitrite-evoked synergistic death of glucose-deprived astrocytes

Chung Ju, Young J. Oh, Byung H. Han, Hee Sun Kim, Hyoung Chun Kim, Won-Ki Kim

Research output: Contribution to journalArticle

6 Citations (Scopus)

Abstract

Previously, we reported that glucose-deprived astrocytes were highly vulnerable to peroxynitrite (ONOO -). Here we demonstrate that the increased vulnerability caused by glucose deprivation and ONOO - depends on intracellular pH. The ONOO - releasing reagent 3-morpholinosydnonimine (SIN-1) markedly induced the release of lactate dehydrogenase (LDH, the marker of cytotoxicity) in glucose-deprived astrocytes. Morphological studies and caspase activity assay showed that astrocytes treated together with glucose deprivation and ONOO - died mostly in a necrotic mode. Alkalinization of pH from 7.4 to 7.8 increased LDH release, whereas acidification from pH 7.4 to 7.0 decreased it. However, intracellular pH (pH i), not extracellular pH (pH e), appeared to play a critical role in the synergistic death. Thus, without a change in pH e (7.4) cytosolic acidification by a weak acid salt, sodium acetate, and a Na +/H + antiporter inhibitor, amiloride, reduced LDH release. In contrast, a weak base, NH 4Cl, and a Na +/H + antiporter stimulator, monensin, increased pH i and greatly enhanced LDH release. The augmented death was found to be due, in part, to the preceding decrease in the level of reduced glutathione, the ONOO - scavenger, and collapse of the mitochondrial transmembrane potential at alkaline pH.

Original languageEnglish
Pages (from-to)1160-1169
Number of pages10
JournalFree Radical Biology and Medicine
Volume37
Issue number8
DOIs
Publication statusPublished - 2004 Oct 15
Externally publishedYes

Fingerprint

Peroxynitrous Acid
Astrocytes
Glucose
Sodium-Hydrogen Antiporter
Acidification
Sodium Acetate
Monensin
Amiloride
Cytotoxicity
Caspases
L-Lactate Dehydrogenase
Glutathione
Assays
Salts
Acids
Membrane Potentials

Keywords

  • Astrocytes
  • Free radical
  • Mitochondrial permeability transition
  • Mitochondrial transmembrane permeability (Δψ )
  • Peroxynitrite (ONOO )
  • pH

ASJC Scopus subject areas

  • Medicine(all)
  • Toxicology
  • Clinical Biochemistry

Cite this

Intracellular pH-dependent peroxynitrite-evoked synergistic death of glucose-deprived astrocytes. / Ju, Chung; Oh, Young J.; Han, Byung H.; Kim, Hee Sun; Kim, Hyoung Chun; Kim, Won-Ki.

In: Free Radical Biology and Medicine, Vol. 37, No. 8, 15.10.2004, p. 1160-1169.

Research output: Contribution to journalArticle

Ju, Chung ; Oh, Young J. ; Han, Byung H. ; Kim, Hee Sun ; Kim, Hyoung Chun ; Kim, Won-Ki. / Intracellular pH-dependent peroxynitrite-evoked synergistic death of glucose-deprived astrocytes. In: Free Radical Biology and Medicine. 2004 ; Vol. 37, No. 8. pp. 1160-1169.
@article{7941db01cda64ab0b2dee15d5ba52a39,
title = "Intracellular pH-dependent peroxynitrite-evoked synergistic death of glucose-deprived astrocytes",
abstract = "Previously, we reported that glucose-deprived astrocytes were highly vulnerable to peroxynitrite (ONOO -). Here we demonstrate that the increased vulnerability caused by glucose deprivation and ONOO - depends on intracellular pH. The ONOO - releasing reagent 3-morpholinosydnonimine (SIN-1) markedly induced the release of lactate dehydrogenase (LDH, the marker of cytotoxicity) in glucose-deprived astrocytes. Morphological studies and caspase activity assay showed that astrocytes treated together with glucose deprivation and ONOO - died mostly in a necrotic mode. Alkalinization of pH from 7.4 to 7.8 increased LDH release, whereas acidification from pH 7.4 to 7.0 decreased it. However, intracellular pH (pH i), not extracellular pH (pH e), appeared to play a critical role in the synergistic death. Thus, without a change in pH e (7.4) cytosolic acidification by a weak acid salt, sodium acetate, and a Na +/H + antiporter inhibitor, amiloride, reduced LDH release. In contrast, a weak base, NH 4Cl, and a Na +/H + antiporter stimulator, monensin, increased pH i and greatly enhanced LDH release. The augmented death was found to be due, in part, to the preceding decrease in the level of reduced glutathione, the ONOO - scavenger, and collapse of the mitochondrial transmembrane potential at alkaline pH.",
keywords = "Astrocytes, Free radical, Mitochondrial permeability transition, Mitochondrial transmembrane permeability (Δψ ), Peroxynitrite (ONOO ), pH",
author = "Chung Ju and Oh, {Young J.} and Han, {Byung H.} and Kim, {Hee Sun} and Kim, {Hyoung Chun} and Won-Ki Kim",
year = "2004",
month = "10",
day = "15",
doi = "10.1016/j.freeradbiomed.2004.07.011",
language = "English",
volume = "37",
pages = "1160--1169",
journal = "Free Radical Biology and Medicine",
issn = "0891-5849",
publisher = "Elsevier Inc.",
number = "8",

}

TY - JOUR

T1 - Intracellular pH-dependent peroxynitrite-evoked synergistic death of glucose-deprived astrocytes

AU - Ju, Chung

AU - Oh, Young J.

AU - Han, Byung H.

AU - Kim, Hee Sun

AU - Kim, Hyoung Chun

AU - Kim, Won-Ki

PY - 2004/10/15

Y1 - 2004/10/15

N2 - Previously, we reported that glucose-deprived astrocytes were highly vulnerable to peroxynitrite (ONOO -). Here we demonstrate that the increased vulnerability caused by glucose deprivation and ONOO - depends on intracellular pH. The ONOO - releasing reagent 3-morpholinosydnonimine (SIN-1) markedly induced the release of lactate dehydrogenase (LDH, the marker of cytotoxicity) in glucose-deprived astrocytes. Morphological studies and caspase activity assay showed that astrocytes treated together with glucose deprivation and ONOO - died mostly in a necrotic mode. Alkalinization of pH from 7.4 to 7.8 increased LDH release, whereas acidification from pH 7.4 to 7.0 decreased it. However, intracellular pH (pH i), not extracellular pH (pH e), appeared to play a critical role in the synergistic death. Thus, without a change in pH e (7.4) cytosolic acidification by a weak acid salt, sodium acetate, and a Na +/H + antiporter inhibitor, amiloride, reduced LDH release. In contrast, a weak base, NH 4Cl, and a Na +/H + antiporter stimulator, monensin, increased pH i and greatly enhanced LDH release. The augmented death was found to be due, in part, to the preceding decrease in the level of reduced glutathione, the ONOO - scavenger, and collapse of the mitochondrial transmembrane potential at alkaline pH.

AB - Previously, we reported that glucose-deprived astrocytes were highly vulnerable to peroxynitrite (ONOO -). Here we demonstrate that the increased vulnerability caused by glucose deprivation and ONOO - depends on intracellular pH. The ONOO - releasing reagent 3-morpholinosydnonimine (SIN-1) markedly induced the release of lactate dehydrogenase (LDH, the marker of cytotoxicity) in glucose-deprived astrocytes. Morphological studies and caspase activity assay showed that astrocytes treated together with glucose deprivation and ONOO - died mostly in a necrotic mode. Alkalinization of pH from 7.4 to 7.8 increased LDH release, whereas acidification from pH 7.4 to 7.0 decreased it. However, intracellular pH (pH i), not extracellular pH (pH e), appeared to play a critical role in the synergistic death. Thus, without a change in pH e (7.4) cytosolic acidification by a weak acid salt, sodium acetate, and a Na +/H + antiporter inhibitor, amiloride, reduced LDH release. In contrast, a weak base, NH 4Cl, and a Na +/H + antiporter stimulator, monensin, increased pH i and greatly enhanced LDH release. The augmented death was found to be due, in part, to the preceding decrease in the level of reduced glutathione, the ONOO - scavenger, and collapse of the mitochondrial transmembrane potential at alkaline pH.

KW - Astrocytes

KW - Free radical

KW - Mitochondrial permeability transition

KW - Mitochondrial transmembrane permeability (Δψ )

KW - Peroxynitrite (ONOO )

KW - pH

UR - http://www.scopus.com/inward/record.url?scp=4644278350&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=4644278350&partnerID=8YFLogxK

U2 - 10.1016/j.freeradbiomed.2004.07.011

DO - 10.1016/j.freeradbiomed.2004.07.011

M3 - Article

C2 - 15451056

AN - SCOPUS:4644278350

VL - 37

SP - 1160

EP - 1169

JO - Free Radical Biology and Medicine

JF - Free Radical Biology and Medicine

SN - 0891-5849

IS - 8

ER -