Impairment of Endothelial Cell Function Induced by Hemoglobin A1c and the Potential Mechanisms

J. Bo; Y. Guan; Y. Guo; S. Xie; C. Zhang; H. Zhang; Z. Chen; J. Lu; Q. H. Meng

doi:10.1055/s-0035-1554622

Experimental and Clinical Endocrinology & Diabetes, Inhaltsverzeichnis

Exp Clin Endocrinol Diabetes 2015; 123(09): 529-535
DOI: 10.1055/s-0035-1554622

Article

Impairment of Endothelial Cell Function Induced by Hemoglobin A_1c and the Potential Mechanisms

Autor*innen

J. Bo

¹Key Laboratory of Laboratory Medicine, Ministry of Education of China, Zhejiang Provincial Key Laboratory of Medical Genetics, School of Laboratory Medicine and Life Sciences, Wenzhou Medical University, Wenzhou, China
Y. Guan

¹Key Laboratory of Laboratory Medicine, Ministry of Education of China, Zhejiang Provincial Key Laboratory of Medical Genetics, School of Laboratory Medicine and Life Sciences, Wenzhou Medical University, Wenzhou, China
Y. Guo

¹Key Laboratory of Laboratory Medicine, Ministry of Education of China, Zhejiang Provincial Key Laboratory of Medical Genetics, School of Laboratory Medicine and Life Sciences, Wenzhou Medical University, Wenzhou, China
S. Xie

¹Key Laboratory of Laboratory Medicine, Ministry of Education of China, Zhejiang Provincial Key Laboratory of Medical Genetics, School of Laboratory Medicine and Life Sciences, Wenzhou Medical University, Wenzhou, China
C. Zhang

¹Key Laboratory of Laboratory Medicine, Ministry of Education of China, Zhejiang Provincial Key Laboratory of Medical Genetics, School of Laboratory Medicine and Life Sciences, Wenzhou Medical University, Wenzhou, China
H. Zhang

¹Key Laboratory of Laboratory Medicine, Ministry of Education of China, Zhejiang Provincial Key Laboratory of Medical Genetics, School of Laboratory Medicine and Life Sciences, Wenzhou Medical University, Wenzhou, China
Z. Chen

²Department of Laboratory Medicine, the First Affiliated Hospital of Wenzhou Medical College, Wenzhou, China
J. Lu

¹Key Laboratory of Laboratory Medicine, Ministry of Education of China, Zhejiang Provincial Key Laboratory of Medical Genetics, School of Laboratory Medicine and Life Sciences, Wenzhou Medical University, Wenzhou, China
Q. H. Meng

³Department of Laboratory Medicine, The University of Texas MD Anderson Cancer Center, Houston, USA

Abstract

Objective: Hemoglobin A_1c (HbA_1c) concentrations reflect glycemic control and diabetic complications. However, there is little evidence supporting the pathological role of HbA_1c in the development and progression of diabetic complications. We investigated the impact of HbA_1c on endothelial cell function and the potential mechanisms.

Methods: The effects of HbA_1c on the viability and migration of human umbilical vein endothelial cells (HUVECs) were measured by the Cell Counting Kit-8 and a wound healing scratch assay, respectively. Production of nitric oxide (NO) and reactive oxygen species was measured by the nitrate reductase colorimetric method and flow cytometry, respectively. The expression of endothelial nitric oxide synthase (eNOS) mRNA was quantitated by reverse-transcriptase PCR. The expression of eNOS, p-AMPK, and NOX4 proteins was detected by Western blot.

Results: High concentrations of HbA_1c reduced the viability and migration of HUVECs in a dose- and time-dependent manner. High concentrations of HbA_1c inhibited production of NO but increased production of ROS. Incubation with increasing concentrations of HbA_1c downregulated the expression of eNOS mRNA, decreased expression of eNOS and p-AMPK, and upregulated expression of NOX4.

Conclusion: These findings provide direct evidence that HbA_1c is involved in the development and progression of the cardiovascular complications of diabetes.

Key words

diabetes mellitus - hemoglobin A_1c - HUVECs - nitric oxide - reaction oxygen species - signal pathway

Volltext

Referenzen

References
1 Krishnamurti U, Steffes MW. Glycohemoglobin: a primary predictor of the development or reversal of complications of diabetes mellitus. Clinical chemistry 2001; 47: 1157-1165
2 Wild S, Roglic G, Green A et al. Global prevalence of diabetes: estimates for the year 2000 and projections for 2030. Diabetes care 2004; 27: 1047-1053
3 Standards of medical care in diabetes–2012. Diabetes care 2012; 35 (Suppl. 01) S11-S63
4 Rahbar S. The discovery of glycated hemoglobin: a major event in the study of nonenzymatic chemistry in biological systems. Annals of the New York Academy of Sciences 2005; 1043: 9-19
5 Sacks DB. Measurement of hemoglobin A(1c): a new twist on the path to harmony. Diabetes care 2012; 35: 2674-2680
6 Ravid M, Brosh D, Ravid-Safran D et al. Main risk factors for nephropathy in type 2 diabetes mellitus are plasma cholesterol levels, mean blood pressure, and hyperglycemia. Archives of internal medicine 1998; 158: 998-1004
7 Avogaro A, Albiero M, Menegazzo L et al. Endothelial dysfunction in diabetes: the role of reparatory mechanisms. Diabetes care 2011; 34 (Suppl. 02) S285-S290
8 Rajendran P, Rengarajan T, Thangavel J et al. The Vascular Endothelium and Human Diseases. International journal of biological sciences 2013; 9: 1057-1069
9 Vallejo S, Angulo J, Peiro C et al. Highly glycated oxyhaemoglobin impairs nitric oxide relaxations in human mesenteric microvessels. Diabetologia 2000; 43: 83-90
10 Rodriguez-Manas L, Arribas S, Giron C et al. Interference of glycosylated human hemoglobin with endothelium-dependent responses. Circulation 1993; 88: 2111-2116
11 Angulo J, Sanchez-Ferrer CF, Peiro C et al. Impairment of endothelium-dependent relaxation by increasing percentages of glycosylated human hemoglobin. Possible mechanisms involved. Hypertension 1996; 28: 583-592
12 Lu J, Ji J, Meng H et al. The protective effect and underlying mechanism of metformin on neointima formation in fructose-induced insulin resistant rats. Cardiovascular diabetology 2013; 12: 58
13 Mazière C, Floret S, Santus R et al. Impairment of the EGF signaling pathway by the oxidative stress generated with UVA. Free Radical Biology and Medicine 2003; 34: 629-636
14 Kirpichnikov D, Sowers JR. Diabetes mellitus and diabetes-associated vascular disease. Trends in endocrinology and metabolism: TEM 2001; 12: 225-230
15 Avogaro A, Fadini GP, Gallo A et al. de Kreutzenberg S. Endothelial dysfunction in type 2 diabetes mellitus. Nutrition, metabolism, and cardiovascular diseases: NMCD 2006; 16 (Suppl. 01) S39-S45
16 Botker HE, Moller N. ON NO–the continuing story of nitric oxide, diabetes, and cardiovascular disease. Diabetes 2013; 62: 2645-2647
17 Alp NJ, Channon KM. Regulation of endothelial nitric oxide synthase by tetrahydrobiopterin in vascular disease. Arteriosclerosis, thrombosis, and vascular biology 2004; 24: 413-420
18 Hoang HH, Padgham SV, Meininger CJ. L-arginine, tetrahydrobiopterin, nitric oxide and diabetes. Current opinion in clinical nutrition and metabolic care 2013; 16: 76-82
19 Conti V, Russomanno G, Corbi G et al. Adrenoreceptors and nitric oxide in the cardiovascular system. Frontiers in physiology 2013; 4: 321
20 Eid AA, Lee DY, Roman LJ et al. Sestrin 2 and AMPK connect hyperglycemia to Nox4-dependent endothelial nitric oxide synthase uncoupling and matrix protein expression. Molecular and cellular biology 2013; 33: 3439-3460
21 Zhou Y, Yan H, Guo M et al. Reactive oxygen species in vascular formation and development. Oxidative medicine and cellular longevity 2013; 2013: 374963
22 Guzik TJ, Mussa S, Gastaldi D et al. Mechanisms of increased vascular superoxide production in human diabetes mellitus: role of NAD(P)H oxidase and endothelial nitric oxide synthase. Circulation 2002; 105: 1656-1662
23 Bitar MS, Al-Mulla F. ROS constitute a convergence nexus in the development of IGF1 resistance and impaired wound healing in a rat model of type 2 diabetes. Disease models & mechanisms 2012; 5: 375-388
24 Sowers JR. Insulin resistance and hypertension. American journal of physiology. Heart and circulatory physiology 2004; 286: H1597-H1602
25 Newsholme P, Homem De Bittencourt P, O’Hagan C et al. Exercise and possible molecular mechanisms of protection from vascular disease and diabetes: the central role of ROS and nitric oxide. Clinical science (London, England: 1979) 2010; 118: 341-349
26 Inoguchi T, Sonta T, Tsubouchi H et al. Protein kinase C-dependent increase in reactive oxygen species (ROS) production in vascular tissues of diabetes: role of vascular NAD(P)H oxidase. Journal of the American Society of Nephrology: JASN 2003; 14: S227-S232
27 Inoguchi T, Li P, Umeda F et al. High glucose level and free fatty acid stimulate reactive oxygen species production through protein kinase C–dependent activation of NAD(P)H oxidase in cultured vascular cells. Diabetes 2000; 49: 1939-1945
28 Rajagopalan S, Kurz S, Munzel T et al. Angiotensin II-mediated hypertension in the rat increases vascular superoxide production via membrane NADH/NADPH oxidase activation. Contribution to alterations of vasomotor tone. The Journal of clinical investigation 1996; 97: 1916-1923
29 Ushio-Fukai M, Zafari AM, Fukui T et al. p22phox is a critical component of the superoxide-generating NADH/NADPH oxidase system and regulates angiotensin II-induced hypertrophy in vascular smooth muscle cells. The Journal of biological chemistry 1996; 271: 23317-23321
30 Griendling KK, Minieri CA, Ollerenshaw JD et al. Angiotensin II stimulates NADH and NADPH oxidase activity in cultured vascular smooth muscle cells. Circulation research 1994; 74: 1141-1148
31 Mohazzab KM, Kaminski PM, Wolin MS. NADH oxidoreductase is a major source of superoxide anion in bovine coronary artery endothelium. The American journal of physiology 1994; 266: H2568-H2572
32 Petry A, Djordjevic T, Weitnauer M et al. NOX2 and NOX4 mediate proliferative response in endothelial cells. Antioxidants & redox signaling 2006; 8: 1473-1484
33 Wang S, Song P, Zou MH. AMP-activated protein kinase, stress responses and cardiovascular diseases. Clinical science (London, England: 1979) 2012; 122: 555-573
34 Levine YC, Li GK, Michel T. Agonist-modulated regulation of AMP-activated protein kinase (AMPK) in endothelial cells. Evidence for an AMPK -> Rac1 -> Akt -> endothelial nitric-oxide synthase pathway. The Journal of biological chemistry 2007; 282: 20351-20364
35 Wang S, Zhang M, Liang B et al. AMPKalpha2 deletion causes aberrant expression and activation of NAD(P)H oxidase and consequent endothelial dysfunction in vivo: role of 26S proteasomes. Circulation research 2010; 106: 1117-1128
36 Morrow VA, Foufelle F, Connell JM et al. Direct activation of AMP-activated protein kinase stimulates nitric-oxide synthesis in human aortic endothelial cells. The Journal of biological chemistry 2003; 278: 31629-31639
37 Hurley RL, Barre LK, Wood SD et al. Regulation of AMP-activated protein kinase by multisite phosphorylation in response to agents that elevate cellular cAMP. The Journal of biological chemistry 2006; 281: 36662-36672
38 Colombo SL, Moncada S. AMPKalpha1 regulates the antioxidant status of vascular endothelial cells. The Biochemical journal 2009; 421: 163-169