The role of aldose reductase in sugar cataract formation: aldose reductase plays a key role in lens epithelial cell death (apoptosis).
Since aldose reductase is localized primarily in lens epithelial cells, osmotic insults induced by the accumulation of sugar alcohols occur first in these cells. To determine whether the accumulation of sugar alcohols can induce lens epithelial cell death, galactose-induced apoptosis has been investigated in dog lens epithelial cells. Dog lens epithelial cells were cultured in Dulbecco's modified Eagle's mimimum essential medium (DMEM) supplemented with 20% fetal calf serum (FCS). After reaching confluence at fifth passage, the medium was replaced with the same DMEM medium containing 50 mM D-galactose and the cells were cultured for an additional 2 weeks. Almost all of the cells cultured in galactose medium were stained positively for apoptosis with the terminal deoxynucleotidyl transferance-mediated biotin-dUTP nick end labeling (TUNEL) technique. Agarose gel electrophoresis of these cells displayed obvious DNA fragmentation, known as a ladder formation. All of these apoptotic changes were absent in similar cells cultured in galactose medium containing 1 microM of the aldose reductase inhibitor AL 1576. Addition of AL 1576 also reduced the cellular galactitol levels from 123+/-10 microgram/10(6) cells (n=5) to 3.9+/-1.9 microgram/10(6) cells (n=5). These observations confirm that galactose induced apoptosis occurs in dog lens epithelial cells. Furthermore, the prevention of apoptosis by an aldose reductase inhibitor suggests that this apoptosis is linked to the accumulation of sugar alcohols.
Murata
M
M
Laboratory of Ocular Therapeutics, National Eye Institute, National Institutes of Health, Rm 10/10B09, 10 Center Drive, MSC 1850, 20892-1850, Bethesda, MD, USA.
Ohta
N
N
Sakurai
S
S
Alam
S
S
Tsai
J
J
Kador
P F
PF
Sato
S
S
D000449:Aldehyde Reductase / Q000037:antagonists & inhibitors / Q000235:genetics / Q000378:metabolism*; D000818:Animals; D017209:Apoptosis / Q000187:drug effects; D001483:Base Sequence; D002386:Cataract / Q000209:etiology* / Q000378:metabolism* / Q000473:pathology; D002478:Cells, Cultured; D017931:DNA Primers / Q000235:genetics; D004285:Dogs; D004791:Enzyme Inhibitors / Q000494:pharmacology; D004847:Epithelial Cells / Q000166:cytology / Q000187:drug effects / Q000378:metabolism; D005449:Fluorenes / Q000494:pharmacology; D004376:Galactitol / Q000378:metabolism; D005690:Galactose / Q000494:pharmacology; D006827:Hydantoins / Q000494:pharmacology; D007908:Lens, Crystalline / Q000166:cytology* / Q000187:drug effects / Q000378:metabolism*; D013402:Sugar Alcohols / Q000378:metabolism*
D017931:DNA Primers; D004791:Enzyme Inhibitors; D005449:Fluorenes; D006827:Hydantoins; D013402:Sugar Alcohols; C052241:imirestat; D004376:Galactitol; D000449:Aldehyde Reductase; D005690:Galactose