Document Detail


Swelling, acidosis, and irreversible damage of glial cells from exposure to arachidonic acid in vitro.
MedLine Citation:
PMID:  7929645     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
Swelling and damage of C6 glioma cells and of primary cultured astrocytes were analyzed in vitro during incubation with arachidonic acid (AA; 20:4). The cells were suspended in a physiological medium supplemented with AA at concentrations of 0.001-1.0 mM. Cell swelling was quantified by flow cytometry with hydrodynamic focusing. Flow cytometry was also utilized for assessment of cell viability by exclusion of the fluorescent dye propidium iodide and for measurement of the intracellular pH (pHi) by 2',7'-bis-(2-carboxyethyl)-5(and -6)carboxy-fluorescein. Administration of AA caused an immediate dose-dependent swelling of C6 glioma cells, even at a concentration of 0.01 mM. At this level cell volume increased within 20 min to 105.0% of control, at 0.1 mM to 111.0%, while at 1.0 mM to 123.7%. Following a phase of rapid cell volume increase, swelling leveled off during the subsequent observation period of 70 min. Viability of the C6 glioma cells was 90% under control conditions. It remained unchanged after raising AA concentrations to 0.1 mM. At 0.5 mM, however, cell viability fell to 72.8%, and at 1.0 mM to 32.7%. pHi of the glioma cells was 7.3 under control conditions. In parallel with the early swelling phase, AA led to a dose-dependent decrease of the intracellular pH and an elevated lactate production of the cells. During incubation with 0.1 mM AA, pHi decreased to 7.06 after 5 min, but recovered to normal subsequently. In addition, swelling-inducing properties of linoleic (18:2) or stearic (18:0) acid were analyzed for evaluation of the specificity of glial swelling induced by AA. Whereas stearic acid (0.1 mM) failed to induce a swelling response, linoleic acid (0.1 mM) was found to be effective. The volume increase of the glial cells, however, was only half of that found during exposure to AA at the same concentration. Further, glial swelling from AA or linoleic acid was completely inhibited by the aminosteroid U-74389F, an antagonist of lipid peroxidation. Finally, omission of Na+ ions in the suspension medium with replacement by choline led also to inhibition of the cell volume increase by AA. Experiments using astrocytes from primary culture confirmed the swelling-inducing properties of AA at a quantitative level, whereas vulnerability of the cells to AA was increased. The present results demonstrate an important role of AA in cytotoxic swelling and irreversible damage of glial cells at concentrations that occur in vivo in cerebral ischemia or trauma.(ABSTRACT TRUNCATED AT 250 WORDS)
Authors:
F Staub; A Winkler; J Peters; O Kempski; V Kachel; A Baethmann
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Publication Detail:
Type:  Journal Article    
Journal Detail:
Title:  Journal of cerebral blood flow and metabolism : official journal of the International Society of Cerebral Blood Flow and Metabolism     Volume:  14     ISSN:  0271-678X     ISO Abbreviation:  J. Cereb. Blood Flow Metab.     Publication Date:  1994 Nov 
Date Detail:
Created Date:  1994-11-21     Completed Date:  1994-11-21     Revised Date:  2000-12-18    
Medline Journal Info:
Nlm Unique ID:  8112566     Medline TA:  J Cereb Blood Flow Metab     Country:  UNITED STATES    
Other Details:
Languages:  eng     Pagination:  1030-9     Citation Subset:  IM    
Affiliation:
Institute for Surgical Research, Ludwig Maximilians University, Munich, Germany.
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MeSH Terms
Descriptor/Qualifier:
Acidosis / chemically induced*,  pathology*
Arachidonic Acid / pharmacokinetics,  pharmacology*
Astrocytes
Cell Survival
Fatty Acids / pharmacology
Hydrogen-Ion Concentration
Lactates / biosynthesis
Lactic Acid
Neuroglia / drug effects,  metabolism*,  pathology*
Sodium / pharmacology
Steroids / pharmacology
Tumor Cells, Cultured
Chemical
Reg. No./Substance:
0/Fatty Acids; 0/Lactates; 0/Steroids; 50-21-5/Lactic Acid; 506-32-1/Arachidonic Acid; 7440-23-5/Sodium

From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine


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