Document Detail


Involvement of Hsp70, a stress protein, in the resistance of long-term culture of PC12 cells against sodium nitroprusside (SNP)-induced cell death.
MedLine Citation:
PMID:  20559625     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
Sodium nitroprusside (SNP)-treated PC12 cell line is being used in our laboratory as a cell model of nitric oxide (NO)-mediated damage for in vitro evaluation of potential neuroprotective compounds, thus cell response to SNP must be standardized to gain reproducible data. The NO-donor SNP has been shown to induce cell death at high concentrations in undifferentiated PC12 cells. Differences were found in sensitivity to SNP between cells from short- and long-term cultured cells. After 24-h exposure to 100-500 microM SNP, a decrease of cell viability was observed in both short- (17, 21 and 23rd passages) and long-term cultures (46, 49 and 50th passages), with IC(50) values of 312.72 and 462.90 microM, respectively. In cells from early passages, SNP-induced cell death was accompanied by significant increases of LDH leakage, nitrite production, malondialdehyde (MDA) levels, catalase (CAT) activity, cleavage of poly(ADP-ribose)polymerase (PARP) and caspase-3 activation in comparison with those from late passages. Furthermore, untreated and SNP-treated cells from long-term cultures displayed an increase of the stress protein Hsp70 levels when compared with those from short-term cultures. Up-regulated levels of Hsp70 may be associated with cell survival. Therefore, cells may acquire a certain resistance to SNP-induced toxicity associated with an increase in cell passage-dependent Hsp70. The protein Hsp70 might modulate the cellular response to the toxic insult by increasing CAT and GSH-Px activities and decreasing caspase-3 activation. Finally, it is crucial for the standardization of this cell model of neurotoxicity, at least in part, the use of PC12 cells in an optimum and reliable range of passages.
Authors:
Carmen Romero; Juana Benedí; Angel Villar; Sagrario Martín-Aragón
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Publication Detail:
Type:  Journal Article; Research Support, Non-U.S. Gov't     Date:  2010-06-18
Journal Detail:
Title:  Archives of toxicology     Volume:  84     ISSN:  1432-0738     ISO Abbreviation:  Arch. Toxicol.     Publication Date:  2010 Sep 
Date Detail:
Created Date:  2010-09-02     Completed Date:  2010-12-02     Revised Date:  -    
Medline Journal Info:
Nlm Unique ID:  0417615     Medline TA:  Arch Toxicol     Country:  Germany    
Other Details:
Languages:  eng     Pagination:  699-708     Citation Subset:  IM    
Affiliation:
Pharmacology Department, School of Pharmacy, Complutense University of Madrid, 28040 Madrid, Spain.
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MeSH Terms
Descriptor/Qualifier:
Animals
Apoptosis
Caspase 3 / metabolism
Cell Culture Techniques
Cell Death
Cell Survival
HSP70 Heat-Shock Proteins / metabolism*,  pharmacology
Neuroprotective Agents / toxicity*
Nitric Oxide / metabolism,  toxicity*
Nitric Oxide Donors / toxicity
Nitroprusside / toxicity*
PC12 Cells
Poly(ADP-ribose) Polymerases / metabolism
Rats
Chemical
Reg. No./Substance:
0/HSP70 Heat-Shock Proteins; 0/Neuroprotective Agents; 0/Nitric Oxide Donors; 10102-43-9/Nitric Oxide; 15078-28-1/Nitroprusside; EC 2.4.2.30/Poly(ADP-ribose) Polymerases; EC 3.4.22.-/Caspase 3

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


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