Glyoxalase I deficiency is associated with an unusual level of advanced glycation end products in a hemodialysis patient.
BACKGROUND
Advanced glycation of proteins and their attendant advanced glycation end products (AGEs) contribute to the complications associated with diabetes mellitus or uremia. Regulatory mechanisms of AGE formation in vivo remain an issue of particular interest. We investigated a role of the glyoxalase detoxification system of precursor reactive carbonyl compounds (RCOs) in the in vivo AGE formation.


METHODS
Plasma levels of AGEs [pentosidine and Nepsilon-carboxymethyllysine (CML)], their RCO precursors, d-lactate (the final product resulting from the glyoxalase detoxification pathway), as well as of various compounds known to generate AGE precursors and surrogate markers for oxidative stress (antioxidant enzymes and glutathione), were measured in both hemodialysis (HD) patients and normal subjects. The activity and protein expression of glyoxalase I, an enzyme essential for the detoxification of alpha-oxoaldehydes, in red blood cells (RBC) were also examined.


RESULTS
In one 69-year-old lady who had been on hemodialysis (HD) for three years and had suffered from recurrent cardiovascular complications despite the absence of significant risk factors, plasma levels of pentosidine (77.3 +/- 2.4 pmol/mg protein) and CML (330.8 +/- 8.2 pmol/mg protein) were markedly elevated as compared to other HD patients (N = 20: 26.6 +/- 11.8 pmol/mg protein for pentosidine and 224.4 +/- 51.7 pmol/mg protein for CML). The plasma level of RCO precursors for pentosidine and CML was also higher in this patient than in other HD patients. Further investigation disclosed a very low activity in RBC of glyoxalase I (1.5 +/- 0.4 mU/106 RBC), as compared to other HD patients (3.9 +/- 0.6 mU/106 RBC) or normal subjects (4.0 +/- 0.6 mU/106 RBC). The glyoxalase I protein level, assessed in RBC by immunoblot analysis with a specific antibody, was markedly lower than that observed in HD patients and normal subjects. The causes of this deficiency remain unknown. Nucleotide sequencing of the products of reverse transcription-polymerase chain reaction from the patient's mononuclear cells revealed no genetic mutation within the coding region of the glyoxalase I gene. Plasma d-lactate level was also in the lower range (0.18 +/- 0.03 mg/dL) of the values measured in the other HD patients (0.27 +/- 0.09 mg/dL) and normal subjects (0.35 +/- 0.12 mg/dL). The plasma levels of various compounds known to generate AGE precursors (glucose, lipids and ascorbic acid) were either normal or low. The surrogate markers for oxidative stress such as antioxidant enzymes (glutathione peroxidases and superoxide dismutase) and glutathione were all within the range observed in the other HD patients.


CONCLUSIONS
The unusually high levels of AGEs in this patient implicate a deficient glyoxalase detoxification of RCO precursors. The present clinical observation implicates, to our knowledge for the first time, the glyoxalase detoxification system and, in particular, glyoxalase in the actual level of AGEs in a uremic patient.
Miyata
T
T
Molecular and Cellular Nephrology, Institute of Medical Sciences and Department of Medicine, Tokai University School of Medicine, and Bohsei-Hiratsuka Clinic, Kanagawa, Japan. t-miyata@is.icc.u-tokai.ac.jp
van Ypersele de Strihou
C
C
Imasawa
T
T
Yoshino
A
A
Ueda
Y
Y
Ogura
H
H
Kominami
K
K
Onogi
H
H
Inagi
R
R
Nangaku
M
M
Kurokawa
K
K
D000368:Aged; D001483:Base Sequence / Q000235:genetics; D005260:Female; D017127:Glycation End Products, Advanced / Q000097:blood*; D006801:Humans; D008658:Inactivation, Metabolic; D007791:Lactoylglutathione Lyase / Q000172:deficiency* / Q000235:genetics / Q000378:metabolism; D018384:Oxidative Stress; D011498:Protein Precursors / Q000097:blood; D012016:Reference Values; D006435:Renal Dialysis; D014511:Uremia / Q000097:blood / Q000628:therapy
D017127:Glycation End Products, Advanced; D011498:Protein Precursors; D007791:Lactoylglutathione Lyase