Korean J Ophthalmol.  2010 Apr;24(2):101-107. 10.3341/kjo.2010.24.2.101.

Protective Effect of Catechin on Apoptosis of the Lens Epithelium in Rats with N-methyl-N-nitrosourea-induced Cataracts

• Lee SM
• Ko IG
• Kim SE
• Kim DH
• Kang BN

• KMID: 946002
• DOI: http://doi.org/10.3341/kjo.2010.24.2.101

Abstract

PURPOSE
To investigate the effect of catechin on apoptotic cell death in the lens epithelium of rats with cataract. METHODS: Cataract was induced by intraperitoneal injection of 100 mg/kg N-methyl-N-nitrosourea (MNU) to ten day-old Sprague-Dawley rats. The neonatal rats were randomly divided into five groups (n=15 in each group): a control group, and four cataract-induction groups, treated with either 0, 50, 100, 200 mg/kg catechin. We performed slit-lamp biomicroscopic analysis, terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling (TUNEL) assay, Western-blot for Bcl-2 and Bax, and immunohistochemistry for caspase-3. RESULTS: Apoptotic cell death in lens epithelial cells that increased following cataract formation in rats was suppressed by cathechin. CONCLUSIONS: Catechin inhibited cataract-induced apoptotic cell death in the lens epithelium and may prove useful for the prevention of cataract progression.

Keyword

Apoptosis; Cataract; Catechin; Crystalline lens; Epithelial cells; Rats

MeSH Terms

Analysis of Variance
Animals
Animals, Newborn
Apoptosis/*drug effects
Blotting, Western
Caspase 3/metabolism
Cataract/chemically induced/*drug therapy
Catechin/*pharmacology
Immunoenzyme Techniques
In Situ Nick-End Labeling
Lens, Crystalline/*drug effects
Random Allocation
Rats
Rats, Sprague-Dawley

Figure

• 

  Fig. 1 Slit-lamp appearance of lenses in rat eyes. (A) control group, (B) cataract-induction group, (C) cataract-induction and 200 mg/kg catechin group.

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  Fig. 2 Effect of catechin on DNA fragmentation in the lens epithelium induced by cataract. Upper: photomicrographs of terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling (TUNEL)-positive cells in the lens epithelium. (1) control group, (2) cataract-induction group, (3) cataract-induction and 200 mg/kg catechin group. The sections were stained for TUNEL. The scale bar represents 200 mm. Lower: number of TUNEL-positive cells in each group. (A) control group, (B) cataract-induction group, (C) cataract-induction and 50 mg/kg catechin group, (D) cataract-induction and 100 mg/kg catechin group, and (E) cataract-induction and 200 mg/kg catechin group.*p<0.05 - compared to the control group; #p<0.05 - compared to the cataract-induced group.

• 

  Fig. 3 Effect of catechin on caspase-3-expression in the lens epithelium induced by cataracts. Upper: photomicrographs of caspase-3-positive cells in the lens epithelium. (1) control group, (2) cataract-induction group, (3) cataract-induction and 200 mg/kg catechin group. The sections were stained for caspase-3 immunoreactivity (brown). The scale bar represents 50 mm. Lower: number of caspase-3-positive cells in each group. (A) control group, (B) cataract-induction group, (C) cataract-induction and 50 mg/kg catechin group, (D) cataract-induction and 100 mg/kg catechin group, and (E) cataract-induction and 200 mg/kg catechin group. *p<0.05 - compared to the control group; #p<0.05 - compared to the cataract-induced group.

• 

  Fig. 4 Effect of catechin on the expressions of Bax and Bcl-2 proteins. Actin was used as an internal control (46 kDa). (A) control group, (B) cataract-induction group, (C) cataract-induction and 50 mg/kg catechin group, (D) cataract-induction and 100 mg/kg catechin group, and (E) cataract-induction and 200 mg/kg catechin group. Upper: the result of band detection using the enhanced chemiluminescence (ECL) detection kit. Lower: (1) The relative expression of Bax protein. (2) The relative expression of Bcl-2 protein. (3) The ratio of Bax to Bcl-2.*p<0.05 - compared to the control group; #p<0.05 - compared to the cataract-induced group.

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