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Abstract #56894 Published in IGR 16-2
Cell senescence reduced the mechanotransduction sensitivity of porcine angular aqueous plexus cells to elevation of pressure
Lei Y; Stamer WD; Wu J; Sun XInvestigative Ophthalmology and Visual Science 2014; 55: 2324-2328
PURPOSE: To investigate the effect of pressure/flow on the barrier function and protein expression of normal and senescent porcine aqueous humor plexus (AAP) cells, which are the porcine equivalent of human Schlemm's canal endothelial cells. METHODS: AAP cells were grown for 2 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 mm Hg for 72 hours. Hydraulic conductivity (HC) and transendothelial electric resistance (TEER) were measured. The expressions of senescence-associated β-galactosidase and DNA damage marker 8-hydroxy-2'-deoxyguanosine (8-OHdG) were monitored, and the protein expression profile was analyzed 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 (from 1.37 ± 0.12 to 1.64 ± 0.18 μL/mm Hg/min/cm(2), P < 0.05), but not in senescent cells (1.15 ± 0.17 and 1.08 ± 0.10 μL μL/mm Hg/min/cm(2)). TEER changes were consistent with the HC results. Western blot analysis showed that the expression level of myosin light chain, claudin-5, and VE-cadherin significantly reduced under pressure elevation 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.
Full article
Classification:
3.6 Cellular biology (Part of: 3 Laboratory methods)
2.5.2 Schlemms canal (Part of: 2 Anatomical structures in glaucoma > 2.5 Meshwork)
3.5 Molecular biology incl. SiRNA (Part of: 3 Laboratory methods)
5.3 Other (Part of: 5 Experimental glaucoma; animal models)
