Upregulation of glyoxalase I fails to normalize methylglyoxal levels: a possible mechanism for biochemical changes in diabetic mouse lenses.
Glyoxalase I is the first enzyme in a two-enzyme glyoxalase system that metabolizes physiological methylglyoxal (MGO). MGO reacts with proteins to form irreversible adducts that may lead to crosslinking and aggregation of lens proteins in diabetes. This study examined the effect of hyperglycemia on glyoxalase I activity and its mRNA content in mouse lens epithelial cells (mLE cells) and in diabetic mouse lenses and investigated the relationship between GSH and MGO in organ cultured lenses. mLE cells cultured with 25 mM D-glucose (high glucose) showed an upregulation of glyoxalase I activity and a higher content of glyoxalase I mRNA when compared with either cells cultured with 5 mM glucose (control) or with 20 mM L-glucose + 5 mM D-glucose. MGO concentration was significantly elevated in cells cultured with high D-glucose, but not in L-glucose. GSH levels were lower in cells incubated with high glucose compared to control cells. Glyoxalase I activity and mRNA levels were elevated in diabetic lenses compared to non-diabetic control mouse lenses. MGO levels in diabetic lenses were higher than in control lenses. Incubation of lenses with buthionine sulfoximine (BSO) resulted in a dramatic decline in GSH but the MGO levels were similar to lenses incubated without BSO. Our data suggest that in mouse lenses MGO accumulation may occur independent of GSH concentration and in diabetes there is an upregulation of glyoxalase I, but this upregulation is inadequate to normalize MGO levels, which could lead to MGO retention and chemical modification of proteins.
Department of Ophthalmology, Case Western Reserve University School of Medicine, Cleveland, OH 44106, USA.
Magdalena M
MM
Staniszewska
Ram H
RH
Nagaraj
D000818:Animals; D003921:Diabetes Mellitus, Experimental / Q000201:enzymology*; D004847:Epithelial Cells / Q000166:cytology / Q000378:metabolism; D005260:Female; D005947:Glucose / Q000378:metabolism / Q000494:pharmacology; D005978:Glutathione / Q000378:metabolism; D007791:Lactoylglutathione Lyase / Q000378:metabolism*; D007908:Lens, Crystalline / Q000201:enzymology*; D051379:Mice; D008807:Mice, Inbred BALB C; D011765:Pyruvaldehyde / Q000378:metabolism*; D012333:RNA, Messenger / Q000378:metabolism; D013997:Time Factors; D015854:Up-Regulation
D012333:RNA, Messenger; D011765:Pyruvaldehyde; D007791:Lactoylglutathione Lyase; D005978:Glutathione; D005947:Glucose