ABSTRACT
<p><b>OBJECTIVE</b>To investigate the role of caspase-3 and its inhibitor Ac-DEVD-CHO in rat lens epithelial cell apoptosis induced by hydrogen peroxide (H(2)O(2)) in vitro.</p><p><b>METHODS</b>Rat lenses were incubated in modified Eagle's medium containing 2 mmol/L H(2)O(2) to induce apoptosis in vitro. Apoptosis in lens epithelial cells was assessed by transmission electron microscopy and annexin V-propidium iodide (PI) double staining flow cytometry after 12, 24 and 48 h of incubation. The activity of caspase-3 was analyzed by western blotting.</p><p><b>RESULTS</b>Observations under transmission electron microscopy revealed that 2 mmol/L H(2)O(2) could effectively induce lens epithelial cell apoptosis in vitro. Caspase-3 activity increased during cell apoptosis and the peak measurement occurred at 24 h after treatment with H(2)O(2). Cell apoptosis was blocked by caspase-3 inhibitor Ac-DEVD-CHO.</p><p><b>CONCLUSIONS</b>The activation of caspase-3 plays an important role in executing apoptosis in H(2)O(2)-treated lens epithelial cells and in the formation of cataract. The caspase-3 inhibitor Ac-DEVD-CHO may effectively prevent lens epithelial cell apoptosis caused by oxidative injury.</p>
Subject(s)
Animals , Female , Rats , Apoptosis , Caspase 3 , Caspase Inhibitors , Caspases , Metabolism , Physiology , Cysteine Proteinase Inhibitors , Pharmacology , Enzyme Activation , Epithelial Cells , Cell Biology , Hydrogen Peroxide , Pharmacology , Lens, Crystalline , Cell Biology , Oligopeptides , Pharmacology , Organ Culture Techniques , Rats, Sprague-DawleyABSTRACT
<p><b>OBJECTIVE</b>To detect the expression of basic fibroblast growth factor (bFGF) in human ocular tissues, and to assess the effect of bFGF on the proliferation of human cataract lens epithelial cells (LECs) and its correlation with age.</p><p><b>METHODS</b>Enucleated eyes were subjected to immunostaining for bFGF protein. Human cataract LECs were cultured in vitro, and treated with bFGF for 48 hr. Proliferation was estimated by the positive area ratio of proliferating cell nuclear antigen (PCNA) in immunohistochemistry.</p><p><b>RESULTS</b>bFGF protein was found in various human ocular tissues. bFGF stimulated human cataract LEC proliferation, and there was an age-related decrease in responsiveness of human cataract LECs to bFGF (P < 0.05).</p><p><b>CONCLUSION</b>bFGF might play an important role in the proliferation of residual human cataract LECs after cataract surgery.</p>