Increased glutathionyl hemoglobin in diabetes mellitus and hyperlipidemia demonstrated by liquid chromatography/electrospray ionization-mass spectrometry.
BACKGROUND
Erythrocytes contain a large amount of glutathione (GSH), which protects cells from oxidative injury. The purpose of this study was to examine whether hemoglobin (Hb) is modified with glutathione by oxidation of the thiol groups in diabetes mellitus and hyperlipidemia, and to determine the oxygen affinity of glutathionyl Hb.


METHODS
Hb samples obtained from patients with type 2 diabetes, patients with hyperlipidemia, and healthy subjects were analyzed by liquid chromatography/electrospray ionization-mass spectrometry (LC/ESI-MS). Glutathionyl Hb was synthesized in vitro by incubating Hb with GSH. The oxygen affinity of glutathionyl Hb was determined by measuring its oxygen dissociation curve.


RESULTS
We first demonstrated that the concentration of glutathionyl Hbbeta chains is markedly increased in the diabetic patients and hyperlipidemic patients compared with healthy subjects. The in vitro synthesis of glutathionyl Hb by incubation of Hb with GSH was enhanced by adding H(2)O(2), a reactive oxygen species, into the incubation solution. The glutathionyl Hb prepared in vitro by incubating Hb with GSH showed a marked increase in oxygen affinity and a marked decrease in the Hill coefficient compared with Hb incubated without GSH.


CONCLUSIONS
Glutathionyl Hb may be useful as a clinical marker of oxidative stress. The increased concentrations of glutathionyl Hb with high oxygen affinity and low cooperativity in diabetes and hyperlipidemia may lead to reduced tissue oxygen delivery.
Niwa
T
T
Nagoya University Daiko Medical Center, 1-1-20 Daiko-minami, Higashi-ku, Nagoya 461-0047, Japan. tniwa@med.nagoyau.ac.jp
Naito
C
C
Mawjood
A H
AH
Imai
K
K
D002853:Chromatography, Liquid; D003920:Diabetes Mellitus / Q000097:blood*; D005260:Female; D005978:Glutathione / Q000737:chemistry / Q000378:metabolism*; D006454:Hemoglobins / Q000737:chemistry / Q000378:metabolism*; D006801:Humans; D006949:Hyperlipidemias / Q000097:blood*; D008297:Male; D013058:Mass Spectrometry; D010100:Oxygen / Q000737:chemistry
D006454:Hemoglobins; C404464:glutathionyl hemoglobin; D005978:Glutathione; D010100:Oxygen