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L. Lu, K. Komeima, S. Usui, P. A. Campochiaro; Increased Expression of the Long Form of Glutathione Peroxidase 4 Strongly Protects Against Oxidative Damage in Cultured RPE Cells. Invest. Ophthalmol. Vis. Sci. 2007;48(13):2933.
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© ARVO (1962-2015); The Authors (2016-present)
To investigate the effect of overexpression of glutathione peroxidase 1 (GPx1), GPx4 (long [L] and short [S] forms), or superoxide dysmutase (SOD) 1 and 2 in cultured RPE cells.
ARPE 19 cells were transiently transfected with an expression plasmid carrying the gene GPx1, L-GPx4, S- L-GPx4, SOD1 or SOD2. Transfected cells were treated with various oxidants and protein carbonyl adducts were measured by ELISA.
The process of transfection caused oxidative damage demonstrated by increased protein carbonyl adducts in ARPE-19 cells transfected with empty plasmid or those containing expression constructs for SODs; this increase in carbonyl adducts was not seen in cells transfected with GPx plasmids. Despite the basal increase in carbonyl adducts, cells overexpressing one of the SODs showed reduced oxidant-induced increases in carbonyl adducts compared to control cells, but over-expression of one of the GPxs was more protective with the L-form of GPx4 having the greatest effect. After co-transfection with L-GPx4 and SOD plasmids, there was a basal increase in carbonyl adducts, but strong protection against oxidant-induced increases.
Increased expression of L-GPx4 in cultured RPE cells provides good protection from oxidative damage to proteins. Additional studies are needed to test the effect of increased expression of L-GPx4 alone and in combination with SODs in animal models of oxidative damage in the retina and/or RPE.
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