The Evaluation of Expression of 1–Cys Peroxiredoxin in Cataract

Abstract

Abstract: : Purpose: We evaluated expression of 1–cys Prx in cultured human lens epithelial cells after treating with TGFß or H₂O₂ and as well as in humane cataract. We compared the expression of 1–cys Prx in animal lens organ culture model which was treated with each two materials, TGFß or H₂O₂ , for induction of cataract. Methods: HLE B3 cells, human epithelial cell lines, were cultured. The serum starved cells were incubated with TGFß or H₂O₂ in dose dependent manner. To evaluate the expression of 1–cys Prx mRNA and protein, western and northern blot were performed. Sixty capsulorhexis specimens were obtained at the time of cataract surgery from patients with anterior subcapsular(ASC), nuclear sclerosis(NS), cortical spokes(CS), posterior subcapsular(PSC), and white mature cataract(WC). For ex vivo lens culture, eyes of 8–week–old Sprague–Dawley male rats were enucleated, and 10ng/ml of TGFß or 1mM H₂O₂ were added. The lens capsule was obtained at day 5 after treatment. Protein isolated from lens capsules were transferred to nitrocellulose membranes, and western blot analysis was done. Results: Both the expression of 1–cys Prx mRNA and protein were increased in B3 cell after TGFß exposure up to 5.0ng/ml, then slightly were decreased at 7.5ng/ml. Both 1–cys Prx mRNA and protein were expressed and increased at the level of 200ng/ml H₂O_2, then the increasing tendency was slowing down. The samples of CS and NS cataract showed a similar pattern of 1–cys Prx expression with clear lens. The 1–cys Prx expression was decreased statistically significantly in WC and ASC. In tne ex vivo rats lens model for examination of the effects of TGFß or H₂O₂ on the lens, the lenses which were incubated with 10ng TGFß or 1mM H₂O₂ developed cataract in similar patterns with ASC and WC, respectively. At the evaluation of lens capsule from ex vivo rat lens, the expression of 1–cys Prx protein in lens epithelial cell of TGFß or H₂O₂ treated lenses was reduced statistically significantly in comparison with control. More marked reduction was found in H₂O₂ treated lenses. Conclusions: TGFß and H₂O₂ induced the expression of 1–cys Prx mRNA and protein in vitro model of human lens epithelial cell. However, the cataract induced by the two of materials characterized the reduction of expression of 1–cys Prx protein, and also human cataract model showed a similar pattern of 1–cys–Prx expression. From the present study,we could assume that there was a relation between the expression of 1–cys Prx and cataract prevention, and 1–cys Prx would be considered to a regulatory factor for prevention of cataract.

Keywords: cataract • gene/expression • oxidation/oxidative or free radical damage