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Journal of Zhejiang University SCIENCE A

ISSN 1673-565X(Print), 1862-1775(Online), Monthly

Expression and proteolytic activity of calpain in lens epithelial cells of oxidative cataract

Abstract: Objective: To study the role of calpain in the mechanism of oxidative cataract through detecting the level of intracellular free Ca2+, the expression and proteolytic activity of calpain in the lens epithelial cells (LECs) of H2O2-induced cataract. Methods: Rat lenses were cultured in vitro and cataract was induced by H2O2. The level of intracellular free Ca2+was measured by fluorescence determination with fura-2/AM. The expression of m-calpain protein in LECs was detected with immunohistochemical method. The proteolytic activity in LECs was measured using a fluorogenic synthetic substrate. Results: There were significant differences of the level of intracellular free Ca2+ (P=0.001, 0.000, 0.000), the expression of m-calpain (P=0.001, 0.000, 0.000) and the proteolytic activity of calpain (P=0.001, 0.000, 0.000) between H2O2-induced and control group at 6, 12 and 24 h, respectively. Conclusions: H2O2 can increase intracellular free Ca2+, then enhance the expression and proteolytic activity of calpain which may play a role in the mechanism of oxidative cataract of rat.

Key words: Cataract, Lens epithelial cell, Hydrogen peroxide, Calpain, Calcium


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DOI:

10.1631/jzus.2004.0743

CLC number:

R776

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Received:

2003-01-16

Revision Accepted:

2003-04-21

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