Nitrosylated Proteins in Monocytes as a new Marker of Oxidative-Nitrosative Stress in Diabetic Subjects with Macroangiopathy

Abstract

Background: Peroxynitrite plays an important role in the pathogenesis of diabetic complications. Nitrosylated protein expression in peripheral blood monocytes reflects intracellular peroxynitrite injury, and thus could be a marker of higher diagnostic and prognostic value than plasma nitrotyrosine level. The purpose of this pilot study was to assess if peripheral blood monocytes of diabetic subjects accumulate nitrosylated proteins, and if nitrosylated protein expression correlates with blood glucose control, variables of lipid profile, C-reactive protein concentration (a marker of inflammation), and differs in patients with and without diabetic macrovascular and microvascular complications.

Methods: Nitrosylated protein expression in peripheral blood monocytes (Western blot analysis) was assessed in 31 subjects with diabetes mellitus (29 Type 2, 2 Type 1; 20 males, 11 females; mean age 66 years). The presence of microangiopathy was defined by retinopathy, albumin excretion, and/or neuropathy, and macroangiopathy by carotid plaques, a history of myocardial infarction, and/or stroke.

Results: Diabetic subjects accumulated significant amounts of nitrosylated proteins in peripheral blood monocytes. Nitrosylated protein expression positively correlated with body weight, blood glucose, HbA₁C, and plasma C-reactive protein concentrations in the whole cohort as well as in subjects with diabetic macroangiopathy.

Conclusions: Monocyte nitrosylated protein expression is a new biomarker of metabolic control and inflammation in diabetic subjects with macroangiopathy. A more detailed assessment of diabetic microvascular complications in a larger group of patients is needed to determine if this variable can be employed as a biomarker of the presence, severity, and progression of diabetic neuropathy, retinopathy, and nephropathy.

Key words

nitrosylated proteins - diabetes mellitus - plasma glucose - HbA1C - lipids

References

• 1 Pacher P, Beckman JS, Liaudet L. Nitric oxide and peroxynitrite in health and disease.  Physiol Rev. 2007;  87 315-424
• 2 Szabo C, Mabley JG, Moeller SM, Shimanovich R, Pacher P, Virag L, Soriano FG, Duzer JH Van, Williams W, Salzman AL, Groves JT. Part I: pathogenetic role of peroxynitrite in the development of diabetes and diabetic vascular complications: studies with FP15, a novel potent peroxynitrite decomposition catalyst.  Mol Med. 2002;  8 571-580
• 3 Virag L, Szabo E, Gergely P, Szabo C. Peroxynitrite-induced cytotoxicity: mechanism and opportunities for intervention.  Toxicol Lett. 2003;  140–141 113-124
• 4 Pacher P, Obrosova IG, Mabley JG, Szabo C. Role of nitrosative stress and peroxynitrite in the pathogenesis of diabetic complications. Emerging new therapeutical strategies.  Curr Med Chem. 2005;  12 267-275
• 5 Obrosova IG, Pacher P, Szabo C, Zsengeller Z, Hirooka H, Stevens MJ, Yorek MA. Aldose reductase inhibition counteracts oxidative-nitrosative stress and poly(ADP-ribose) polymerase activation in tissue sites for diabetes complications.  Diabetes. 2005;  54 234-242
• 6 Kowluru RA, Kanwar M, Kennedy A. Metabolic memory phenomenon and accumulation of peroxynitrite in retinal capillaries.  Exp Diabetes Res. 2007;  2007 21976
• 7 Drel VR, Pacher P, Stevens MJ, Obrosova IG. Aldose reductase inhibition counteracts nitrosative stress and poly(ADP-ribose) polymerase activation in diabetic rat kidney and high-glucose-exposed human mesangial cells.  Free Radic Biol Med. 2006;  40 1454-1465
• 8 Obrosova IG, Mabley JG, Zsengeller Z, Charniauskaya T, Abatan OI, Groves JT, Szabo C. Role for nitrosative stress in diabetic neuropathy: evidence from studies with a peroxynitrite decomposition catalyst.  FASEB J. 2005;  19 401-403
• 9 Drel VR, Mashtalir N, Ilnytska O, Shin J, Li F, Lyzogubov VV, Obrosova IG. The leptin-deficient (ob/ob) mouse: a new animal model of peripheral neuropathy of type 2 diabetes and obesity.  Diabetes. 2006;  55 3335-3343
• 10 Vareniuk I, Pavlov IA, Drel VR, Lyzogubov VV, Ilnytska O, Bell SR, Tibrewala J, Groves JT, Obrosova IG. Nitrosative stress and peripheral diabetic neuropathy in leptin-deficient (ob/ob) mice.  Exp Neurol. 2007;  205 425-436
• 11 Drel VR, Pacher P, Vareniuk I, Pavlov I, Ilnytska O, Lyzogubov VV, Tibrewala J, Groves JT, Obrosova IG. A peroxynitrite decomposition catalyst counteracts sensory neuropathy in streptozotocin-diabetic mice.  Eur J Pharmacol. 2007;  569 48-58
• 12 Obrosova IG, Ilnytska O, Lyzogubov VV, Pavlov IA, Mashtalir N, Nadler JL, Drel VR. High-fat diet induced neuropathy of pre-diabetes and obesity: effects of „healthy” diet and aldose reductase inhibition.  Diabetes. 2007;  56 2598-2608
• 13 Drel VR, Pacher P, Vareniuk I, Pavlov IA, Ilnytska O, Lyzogubov VV, Bell SR, Groves JT, Obrosova IG. Evaluation of the peroxynitrite decomposition catalyst Fe(III)tetra-mesitylporphyrin octasulfonate (FeTMPS) on peripheral neuropathy in a mouse model of Type 1 diabetes.  Int J Mol Med. 2007;  20 783-792
• 14 Obrosova IG, Drel VR, Oltman CL, Mashtalir N, Tibrewala J, Groves JT, Yorek MA. Role of nitrosative stress in early neuropathy and vascular dysfunction in streptozotocin-diabetic rats.  Am J Physiol Endocrinol Metab. 2007;  , published online
• 15 Oltman CL, Richou LL, Davidson EP, Coppey LJ, Lund DD, Yorek MA. Progression of coronary and mesenteric vascular dysfunction in Zucker obese and Zucker diabetic fatty rats.  Am J Physiol Heart Circ Physiol. 2006;  291 H1780-H1787
• 16 Ceriello A, Mercuri F, Quagliaro L, Assaloni R, Motz E, Tonutti L, Taboga C. Detection of nitrotyrosine in the diabetic plasma: evidence of oxidative stress.  Diabetologia. 2001;  44 834-838
• 17 Ceriello A, Quagliaro L, Catone B, Pascon R, Piazzola M, Bais B, Marra G, Tonutti L, Taboga C, Motz E. Role of hyperglycemia in nitrotyrosine postprandial generation.  Diabetes Care. 2002;  25 1439-1443
• 18 Ceriello A, Assaloni R, Da Ros R, Maier A, Quagliaro L, Piconi L, Esposito K, Giugliano D. Effect of irbesartan on nitrotyrosine generation in non-hypertensive diabetic patients.  Diabetologia. 2004;  47 1535-1540
• 19 Szabo C, Zanchi A, Komjati K, Pacher P, Krolewski AS, Quist WC, LoGerfo FW, Horton ES, Veves A. Poly(ADP-Ribose) polymerase is activated in subjects at risk of developing type 2 diabetes and is associated with impaired vascular reactivity.  Circulation. 2002;  106 2680-2686
• 20 Frustaci A, Kajstura J, Chimenti C, Jakoniuk I, Leri A, Maseri A, Nadal-Ginard B, Anversa P. Myocardial cell death in human diabetes.  Circ Res. 2000;  87 1123-1132
• 21 Thuraisingham RC, Nott CA, Dodd SM, Yaqoob MM. Increased nitrotyrosine staining in kidneys from patients with diabetic nephropathy.  Kidney Int. 2000;  57 1968-1972
• 22 Nangle MR, Cotter MA, Cameron NE. Effects of the peroxynitrite decomposition catalyst, FeTMPyP, on function of corpus cavernosum from diabetic mice.  Eur J Pharmacol. 2004;  502 143-148
• 23 Zheng L, Du Y, Miller C, Gubitosi-Klug RA, Kern TS, Ball S, Berkowitz BA. Critical role of inducible nitric oxide synthase in degeneration of retinal capillaries in mice with streptozotocin-induced diabetes.  Diabetologia. 2007;  50 1987-1996
• 24 Towbin H, Staehelin T, Gordon J. Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: Procedure and some applications.  Biotechnology. 1992;  24 145-149
• 25 Ceriello A, Piconi L, Esposito K, Giugliano D. Telmisartan shows an equivalent effect of vitamin C in further improving endothelial dysfunction after glycemia normalization in type 1 diabetes.  Diabetes Care. 2007;  30 1694-1698
• 26 Assaloni R, Da Ros R, Quagliaro L, Piconi L, Maier A, Zuodar G, Motz E, Ceriello A. Effects of S21403 (mitiglinide) on postprandial generation of oxidative stress and inflammation in type 2 diabetic patients.  Diabetologia. 2005;  48 1919-1924
• 27 Ceriello A, Lush CW, Darsow T, Piconi L, Corgnali M, Nanayakkara N, Frias JP, Maggs D. Pramlintide reduced markers of oxidative stress in the postprandial period in patients with type 2 diabetes.  Diabetes Metab Res Rev. 2007;  , published online
• 28 Ceriello A, Assaloni R, Da Ros R, Maier A, Piconi L, Quagliaro L, Esposito K, Giugliano D. Effect of atorvastatin and irbesartan, alone and in combination, on postprandial endothelial dysfunction, oxidative stress, and inflammation in type 2 diabetic patients.  Circulation. 2005;  111 2518-2524
• 29 Johnson JB, Summer W, Cutler RG, Martin B, Hyun DH, Dixit VD, Pearson M, Nassar M, Tellejohan R, Maudsley S, Carlson O, John S, Laub DR, Mattson MP. Alternate day calorie restriction improves clinical findings and reduces markers of oxidative stress and inflammation in overweight adults with moderate asthma.  Free Radic Biol Med. 2007;  42 665-674
• 30 Kowluru RA, Odenbach S. Effect of long-term administration of alpha-lipoic acid on retinal capillary cell death and the development of retinopathy in diabetic rats.  Diabetes. 2004;  53 3233-3238
• 31 Ihnat MA, Thorpe JE, Kamat CD, Szabo C, Green DE, Warnke LA, Lacza Z, Cselenyak A, Ross K, Shakir S, Piconi L, Kaltreider RC, Ceriello A. Reactive oxygen species mediate a cellular ‘memory’ of high glucose stress signalling.  Diabetologia. 2007;  50 1523-1531
• 32 Marfella R, D'Amico M, Esposito K, Baldi A, Filippo C Di, Siniscalchi M, Sasso FC, Portoghese M, Cirillo F, Cacciapuoti F, Carbonara O, Crescenzi B, Baldi F, Ceriello A, Nicoletti GF, D'Andrea F, Verza M, Coppola L, Rossi F, Giugliano D. The ubiquitin-proteasome system and inflammatory activity in diabetic atherosclerotic plaques: effects of rosiglitazone treatment.  Diabetes. 2006;  55 622-632
• 33 Hoeldtke RD, Bryner KD, MacNeill DR, Hobbs GR, Baylis C. Peroxynitrite versus nitric oxide in early diabetes.  Am J Hypertens. 2003;  16 ((9 Pt 1)) 761-766
• 34 Vinik AI, Ullal J, Parson HK, Barlow PM, Casellini CM. Pioglitazone treatment improves nitrosative stress in type 2 diabetes.  Diabetes Care. 2006;  29 869-876
• 35 Nourooz-Zadeh J, Ziegler D, Sohr C, Betteridge JD, Knight J, Hothersall J. The use of pholasin as a probe for the determination of plasma total antioxidant capacity.  Clin Biochem. 2006;  39 55-61
• 36 Ziegler D, Sohr CG, Nourooz-Zadeh J. Oxidative stress and antioxi-dant defense in relation to the severity of diabetic polyneuropathy and cardiovascular autonomic neuropathy.  Diabetes Care. 2004;  27 2178-2183
• 37 Hotta N, Toyota T, Matsuoka K, Shigeta Y, Kikkawa R, Kaneko T, Takahashi A, Sugimura K, Koike Y, Ishii J, Sakamoto N. SNK-860 Diabetic Neuropathy Study Group . Clinical efficacy of fidarestat, a novel aldose reductase inhibitor, for diabetic peripheral neuropathy: a 52-week multicenter placebo-controlled double-blind parallel group study.  Diabetes Care. 2001;  24 1776-1782
• 38 Sima AA, Calvani M, Mehra M, Amato A. Acetyl-L-Carnitine Study Group . Acetyl-L-carnitine improves pain, nerve regeneration, and vibratory perception in patients with chronic diabetic neuropathy: an analysis of two randomized placebo-controlled trials.  Diabetes Care. 2005;  28 89-94
• 39 Bril V, Buchanan RA. Long-term effects of ranirestat (AS-3201) on peripheral nerve function in patients with diabetic sensorimotor polyneuropathy.  Diabetes Care. 2006;  29 68-72
• 40 Vinik AI, Tuchman M, Safirstein B, Corder C, Kirby L, Wilks K, Quessy S, Blum D, Grainger J, White J, Silver M. Lamotrigine for treatment of pain associated with diabetic neuropathy: results of two randomized, double-blind, placebo-controlled studies.  Pain. 2007;  128 169-179
• 41 Casellini CM, Barlow PM, Rice AL, Casey M, Simmons K, Pittenger G, Bastyr 3rd EJ, Wolka AM, Vinik AI. A 6-month, randomized, double-masked, placebo-controlled study evaluating the effects of the protein kinase C-beta inhibitor ruboxistaurin on skin microvascular blood flow and other measures of diabetic peripheral neuropathy.  Diabetes Care. 2007;  30 896-902

The authors declare that there are no conflicts of interest.

Correspondence