Document Detail

Oxidative Stress Reduced The Mechanotransduction sensitivity of Porcine Angular Aqueous Plexus Cells To Elevation Of Pressure.
MedLine Citation:
PMID:  24576879     Owner:  NLM     Status:  Publisher    
Purpose:To investigate effect of pressure/flow on the global protein expression profiles of normal and senescent porcine aqueous humour plexus (AAP) cells, which are the porcine equivalent of human Schlemm's canal. Methods:AAP cells were grown for two weeks in physiological (5% O2) or hyperoxic conditions (40% O2) to model cell senescence. Control and senescent AAP cells were subjected to control and elevated hydrostatic pressure gradient of 10 mmHg. Hydraulic conductivity (HC) and transendothelial electric resistance (TEER) was measured. The expression of senescence-associated beta-galactosidase and DNA damage marker 8-hydroxy-2'-deoxyguanosine (8-OHdG) was monitored, and the protein expression profile was analyzed by iTRAQ proteomics method. The expression levels of cytoskeleton and barrier proteins were confirmed by western blot. Results:After 14 days of hyperoxia AAP cells stained positive for 8-OHdG and β-galactosidase. Pressure elevation/flow resulted in significant increase of HC in control cells (1.64 ± 0.18 uL/mmHg/min/cm2, p<0.05), but not in senescent cells (1.34 ± 0.12 uL/mmHg/min/cm2). TEER changes were consistent with the HC results. Proteomics analysis showed that the number of differentially expressed proteins significantly reduced in senescent cells (482 vs. 277 proteins, p<0.05). Cluster of Orthologous Groups analysis further revealed that pressure elevation/flow affected less protein functional groups in senescent cells. The proteins identified by proteomics was verified by western blot analysis, including myosin light chain, claudin-5, ve-cadherin which were significantly reduced in control cells but not in senescent cells. Conclusions:AAP cells are mechano-sensitive, however, cell senescence rendered the cells less responsive to mechanical stimulus which may have pathological consequences.
Yuan Lei; W Daniel Stamer; Jihong Wu; Xinghuai Sun
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Publication Detail:
Type:  JOURNAL ARTICLE     Date:  2014-2-27
Journal Detail:
Title:  Investigative ophthalmology & visual science     Volume:  -     ISSN:  1552-5783     ISO Abbreviation:  Invest. Ophthalmol. Vis. Sci.     Publication Date:  2014 Feb 
Date Detail:
Created Date:  2014-2-28     Completed Date:  -     Revised Date:  -    
Medline Journal Info:
Nlm Unique ID:  7703701     Medline TA:  Invest Ophthalmol Vis Sci     Country:  -    
Other Details:
Languages:  ENG     Pagination:  -     Citation Subset:  -    
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