Biochemical changes in selenite cataract model measured by high-resolution MAS H NMR spectroscopy.
OBJECTIVE
To correlate certain levels of lens opacification with high-resolution magic-angle spinning proton nuclear magnetic resonance (HR-MAS (1)H NMR) spectroscopy analysis of the biochemical changes in rat lenses in a selenite cataract model.


METHODS
Selenite cataract was induced by injecting 13-day-old Sprague-Dawley rat pups with a single subcutaneous dose of sodium selenite (3.28 mg/kg in 0.9% sodium chloride solution). Lens opacification was observed using a photographic slit-lamp microscope at selected time-points 3, 6 and 9 days after selenite injection and was then graded (levels 0, 1 and 2). The animals were killed after the slit-lamp microscopy, lenses were removed and HR-MAS (1)H NMR spectra from intact lenses were obtained. Relative changes in metabolite concentrations were determined after comparison with matched lenses from untreated animals.


RESULTS
Photographic slit-lamp microscopy revealed different stages of cataract in all animals treated with selenite. In the high quality HR-MAS (1)H NMR spectra of the lenses, more than 30 different metabolites were identified in each lens. With the exception of taurine, the concentrations of all amino acids showed a significant increase (p < 0.05) in the second level of cataract. By contrast, glutathione (GSH), succinate and phosphocholine concentrations were significantly reduced.


CONCLUSIONS
For the first time, this study demonstrates the potential to correlate the level of lens opacification with the biochemical changes obtained with HR-MAS (1)H NMR spectroscopy analysis in a selenite cataract model.
Fris
Miroslav
M
Faculty of Medicine, Norwegian University of Science and Technology, Trondheim, Norway. miroslav.fris@ntnu.no
Tessem
May-Britt
MB
Saether
Oddbjørn
O
Midelfart
Anna
A
D000596:Amino Acids / Q000378:metabolism*; D000818:Animals; D000831:Animals, Newborn; D002386:Cataract / Q000139:chemically induced / Q000378:metabolism* / Q000473:pathology; D003459:Crystallins / Q000378:metabolism; D004195:Disease Models, Animal; D005978:Glutathione / Q000378:metabolism*; D007279:Injections, Subcutaneous; D007908:Lens, Crystalline / Q000187:drug effects / Q000378:metabolism*; D019906:Nuclear Magnetic Resonance, Biomolecular; D010767:Phosphorylcholine / Q000378:metabolism; D051381:Rats; D017207:Rats, Sprague-Dawley; D018038:Sodium Selenite / Q000633:toxicity; D019802:Succinic Acid / Q000378:metabolism
D000596:Amino Acids; D003459:Crystallins; D010767:Phosphorylcholine; D019802:Succinic Acid; D005978:Glutathione; D018038:Sodium Selenite