Reactive carbonyls and polyunsaturated fatty acids produce a hydroxyl radical-like species: a potential pathway for oxidative damage of retinal proteins in diabetes.
The pattern of oxidized amino acids in aortic proteins of nonhuman primates suggests that a species resembling hydroxyl radical damages proteins when blood glucose levels are high. However, recent studies argue strongly against a generalized increase in diabetic oxidative stress, which might instead be confined to the vascular wall. Here, we describe a pathway for glucose-stimulated protein oxidation and provide evidence of its complicity in diabetic microvascular disease. Low density lipoprotein incubated with pathophysiological concentrations of glucose became selectively enriched in ortho-tyrosine and meta-tyrosine, implicating a hydroxyl radical-like species in protein damage. Model system studies demonstrated that the reaction pathway requires both a reactive carbonyl group and a polyunsaturated fatty acid, involves lipid peroxidation, and is blocked by the carbonyl scavenger aminoguanidine. To explore the physiological relevance of the pathway, we used mass spectrometry and high pressure liquid chromatography to quantify oxidation products in control and hyperglycemic rats. Hyperglycemia raised levels of ortho-tyrosine, meta-tyrosine, and oxygenated lipids in the retina, a tissue rich in polyunsaturated fatty acids. Rats that received aminoguanidine did not show this increase in protein and lipid oxidation. In contrast, rats with diet-induced hyperlipidemia in the absence of hyperglycemia failed to exhibit increased protein and lipid oxidation products in the retina. Our observations suggest that generation of a hydroxyl radical-like species by a carbonyl/polyunsaturated fatty acid pathway might promote localized oxidative stress in tissues vulnerable to diabetic damage. This raises the possibility that antioxidant therapies that specifically inhibit the pathway might delay the vascular complications of diabetes.
Pennathur
Subramaniam
S
Department of Medicine, University of Washington, Seattle, 98195, USA.
Ido
Yasuo
Y
Heller
Jozsef I
JI
Byun
Jaeman
J
Danda
Ratna
R
Pergola
Pablo
P
Williamson
Joseph R
JR
Heinecke
Jay W
JW
D000596:Amino Acids / Q000737:chemistry; D000818:Animals; D000975:Antioxidants / Q000737:chemistry; D002244:Carbon / Q000737:chemistry*; D002540:Cerebral Cortex / Q000378:metabolism; D002851:Chromatography, High Pressure Liquid; D003920:Diabetes Mellitus / Q000473:pathology*; D003921:Diabetes Mellitus, Experimental / Q000378:metabolism*; D005231:Fatty Acids, Unsaturated / Q000737:chemistry*; D005947:Glucose / Q000378:metabolism*; D006146:Guanidines / Q000737:chemistry; D017665:Hydroxyl Radical; D006943:Hyperglycemia / Q000378:metabolism; D050356:Lipid Metabolism; D015227:Lipid Peroxidation; D008077:Lipoproteins, LDL / Q000378:metabolism; D008297:Male; D013058:Mass Spectrometry; D018384:Oxidative Stress; D010100:Oxygen / Q000737:chemistry / Q000378:metabolism; D010649:Phenylalanine / Q000737:chemistry; D051381:Rats; D017207:Rats, Sprague-Dawley; D012160:Retina / Q000378:metabolism*; D012259:Ribonuclease, Pancreatic / Q000737:chemistry / Q000378:metabolism; D013696:Temperature; D014443:Tyrosine / Q000737:chemistry
D000596:Amino Acids; D000975:Antioxidants; D005231:Fatty Acids, Unsaturated; D006146:Guanidines; D008077:Lipoproteins, LDL; D017665:Hydroxyl Radical; D014443:Tyrosine; D010649:Phenylalanine; D002244:Carbon; D012259:Ribonuclease, Pancreatic; D005947:Glucose; D010100:Oxygen; C004479:pimagedine