Document Detail


Effects of benzo(e)pyrene on the retinal neurosensory cells and human microvascular endothelial cells in vitro.
MedLine Citation:
PMID:  19899995     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
PURPOSE: To study the effects of benzo(e)pyrene (B(e)P), a toxic component of cigarette smoke, on retinal neurosensory (R28) cells and human microvascular endothelial cells (HMVEC). MATERIALS AND METHODS: R28 cells and HMVEC were treated for 24 hours with 1000, 400, 200, and 100 micro M of B(e)P. Cell viability was measured by dye exclusion assay. Caspase-3/7, -8, -9, and -12 activities were measured by fluorochrome assays. DNA ladder was run on agarose gel, and lactate dehydrogenase (LDH) release rate was evaluated using a LDH cytotoxicity kit II. RESULTS: R28 cells exposed to B(e)P 1000 and 400 micro M showed a decrease in cell viability but not at lower concentrations of 200 and 100 micro M. At 400, 200, and 100 micro M B(e)P, there was an increase in caspase-3/7 and at 200 and 100 microM B(e)P caspase-12 activities. Caspase-8 activity was increased only at 200 micro M B(e)P. Caspase-9 activity was not increased at any concentration. DNA ladder revealed bands at 200 bp intervals at lower concentrations and LDH activity at higher concentrations. HMVEC cultures exposed to B(e)P 1000, 400, and 200 micro M showed a decrease in cell viability. Caspase-3/7 activity was not increased at any concentration. DNA laddering revealed no bands at 200 bp intervals at any dose. LDH release rates increased at all three concentrations. However, 100 micro M B(e)P was found safe on HMVEC. CONCLUSIONS: B(e)P has different mechanisms of action on R28 cells and HMVEC at different concentrations. In R28 cells, 200 and 100 micro M of B(e)P causes activation of caspase-3/7, -8 (200 micro M only) and -12 pathways, leading to apoptotic cell death, but, at higher concentrations, there is non-apoptotic cell death, which could be due to necrosis. In contrast, the HMVEC cell death is through non-caspase-dependent necrosis pathway. The molecular mechanisms of cell death vary with different cell types and concentrations of B(e)P.
Authors:
A Jayaprakash Patil; Ana L Gramajo; Ashish Sharma; Marilyn Chwa; Gail M Seigel; Baruch D Kuppermann; M Cristina Kenney
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Publication Detail:
Type:  Journal Article; Research Support, N.I.H., Extramural    
Journal Detail:
Title:  Current eye research     Volume:  34     ISSN:  1460-2202     ISO Abbreviation:  Curr. Eye Res.     Publication Date:  2009 Aug 
Date Detail:
Created Date:  2009-11-10     Completed Date:  2009-12-24     Revised Date:  -    
Medline Journal Info:
Nlm Unique ID:  8104312     Medline TA:  Curr Eye Res     Country:  England    
Other Details:
Languages:  eng     Pagination:  672-82     Citation Subset:  IM    
Affiliation:
Department of Ophthalmology, Gavin S. Herbert Eye Institute, University of California, Irvine, California, USA.
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MeSH Terms
Descriptor/Qualifier:
Animals
Apoptosis / drug effects
Benzopyrenes / toxicity*
Caspases / metabolism
Cell Line
Cell Survival / drug effects
Cells, Cultured
DNA Fragmentation
Endothelium, Vascular / drug effects*,  enzymology,  pathology
Humans
L-Lactate Dehydrogenase / metabolism
Rats
Retina / drug effects*,  enzymology,  pathology
Grant Support
ID/Acronym/Agency:
5R24EY016662/EY/NEI NIH HHS
Chemical
Reg. No./Substance:
0/Benzopyrenes; 192-97-2/benzo(e)pyrene; EC 1.1.1.27/L-Lactate Dehydrogenase; EC 3.4.22.-/Caspases

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


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