Glucose and Insulin Modulate the Capacity of Endothelial Cells (HUVEC) to Express P-selectin and Bind a Monocytic Cell Line (U937)

Summary

Diabetes mellitus is associated with increased prevalence of endothelial cell dysfunction and vascular diseases. Mechanisms leading to alterations in endothelial cell function are poorly understood. We report here that hyperglycaemia results in the expression of endothelial adhesion molecules involved in leukocyte adhesion and extravasation. Incubation of human umbilical cord endothelial cells (HUVEC) with 25 mM glucose induced the expression of P-selectin. This effect was reversed by the addition of 1 nM insulin. Moreover, increased ICAM-1 expression was observed upon HUVEC incubation with 25 mM glucose. Increased adhesion of U937 cells (a monocytic cell line) to endothelial cells cultured with 25 mM glucose was observed. High glucose-induced monocytes cell adhesion was inhibited by an anti-P-selectin monoclonal antibody (LYP20). These results show that high glucose concentration activates endothelial cells leading to monocytes adhesion providing further evidence that hyperglycaemia might be implicated in vessel wall lesions contributing to diabetic vascular disease.

Present address: Dr. M. D. Puente Navazo, Centre Pluridisciplinaire d’Oncologie, ISREC, Epalinges, Switzerland

Keywords

Endothelial cells - P-selectin - hyperglycaemia

• References

• 1 Meyer RY, Herman AG. Vascular endothelial dysfunction. Prog Cardiovasc Dis 1997; 4: 325-42.
• 2 Cosentino F, Luscher TF. Endothelial cell dysfunction in diabetes mellitus. J Cardiovasc Pharmacol 1998; S54-61.
• 3 Sobrevia L, Mann GE. Dysfunction of the endothelial nitric oxide signalling pathway in diabetes and hyperglycaemia. Exp Physiol 1997; 82: 423-52.
• 4 Williams SB, Goldfine AB, Timimi FK, Ting HH, Roddy MA, Simonson DC, Creager MA. Acute hyperglycemia attenuates endothelium-dependent vasodilation in humans in vivo. Circulation 1998; 97: 1695-701.
• 5 Pieper GM. Review of alterations in endothelial nitric oxide production in diabetes: protective role of arginine on endothelial dysfunction. Hypertension 1998; 31: 1047-60.
• 6 Vlassara H. Recents progress in advanced glycation end products and diabetic complications. Diabetes 1997; 32: S19-25.
• 7 Hammes HP, Alt A, Niwa T, Clausen JT, Bretzel RG, Brownlee M, Schleicher ED. Differential accumulation of advanced glycation end products in the course of diabetic retinopathy. Diabetologia 1997; 42: 728-36.
• 8 Yan SD, Stern D, Schmidt AM. What’s the RAGE? The receptor for advanced glycation end products (RAGE) and the dark side of glucose. Eur J Clin Invest. 1997; 2: 179-81.
• 9 Schmidt AM, Hori O, Chen JX, Li JF, Crandall J, Zhang J, Cao R, Yan SD, Brett J, Stern D. Advanced glycosilation endproducts interacting with their endothelial receptor induce the expression of vascular cell adhesion molecule 1 (VCAM-1) in cultured endothelial cells and in mice. J Clin Invest 1995; 96: 1395-403.
• 10 Segawa Y, Shirao Y, Yamagishi S, Higashide T, Kobayashi M, Katsuno K, Iyobe A, Harada H, Sato F, Miyata H, Asai H, Nishimura A, Takahira M, Souno T, Segawa Y, Maeda K, Shima K, Mizuno A, Yamamoto H, Kawasaki K. Upregulation of retinal vascular endothelial growth factor mRNAs in spontaneously diabetic rats without ophthalmoscopic retinopathy: A possible participation of advanced glycation end products in the development of the early phase of diabetic retinopathy. Ophthalmic Research 1998; 30: 333-9.
• 11 Ichikawa K, Yoshinari M, Iwase M, Wakisaka M, Doi Y, Iino K, Yamamoto M, Fujishima M. Advanced glycosylation end products induced tissue factor expression in human monocyte-like U937 cells and increased tissue factor expression in monocytes from diabetic patients. Atherosclerosis. 1998; 136: 281-7.
• 12 Bucala R, Makita Z, Koschinsky T, Cerami A, Vlassara M. Lipid advanced glycosilation: pathway for lipid oxidation in vivo. Proc Natl Acad Sci USA 1993; 90: 6434-8.
• 13 Geburher V, Murphy JF, Bordet JC, Reck M, McGregor JL. Oxidized low-density lipoprotein induces the expression of P-selectin (GMP140/ PADGEM/CD62) on human endothelial cells. Biochem J 1995; 306: 293-8.
• 14 McLeod DS, Lefer DJ, Merges C, Lutty GA. Enhanced expression of intra-cellular adhesion molecule-1 and P-selectin in the diabetic human retina and choroid. Am J Pathol 1995; 147: 642-53.
• 15 Taki H, Kashiwagi A, Tanaka Y, Horrike K. Expression of intercellular adhesion molecules (ICAM-1) via an osmotic effect in human umbilical vein endothelial cells exposed to high glucose medium. Life Sciences. 1996; 58: 1713-21.
• 16 Kim JA, Berliner JA, Natarajan RD, Nadler JL. Evidence that glucose increases monocyte binding to human aortic endothelial cells. Diabetes 1994; 43: 1103-7.
• 17 Kunt T, Forst T, Fruh B, Flohr T, Schneider S, Harzer O, Pfutzner A, Engelbach N, Lobig M. Binding of monocytes from normolipidemic hyper-glycaemic patients with type I diabetes to endothelial cells is increased in vitro. Exp] Clin Endocrin & diabetes 1999; 107: 252-6.
• 18 Lampeter ER, Kishimotto TK, Rothlein R, Mainholfi EA, Bertrans J, Kolb H, Martin S. Elevated levels of circulating adhesion molecules in IDDM patients and in subjects at risk for IDDM. Thromb Haemost 1992; 76: 328-32.
• 19 Fasching P, Veitl M, Rohac M, Streli C, Schneider B, Waldhausl W, Wagner OF. Elevated concentrations of circulating adhesion molecules and their association with microvascular complications in insulin-dependent diabetes mellitus. J Clin Endocrinol Metab 1996; 81: 4313-7.
• 20 Jude EB, Abbot CA, Young MJ, Anderson SG, Douglas JT, Boulton AJ. The potential role of cell adhesion molecules in the pathogenesis of diabetic neuropathy. Diabetologia; 1998; 41: 330-6.
• 21 Parmentier S, McGregor L, Catimel B, Leung LL, McGregor JL. Inhibition of platelet functions by a monoclonal antibody (LYP20) directed against a granule membrane glycoprotein. Blood 1991; 77: 1734-9.
• 22 Puente Navazo MD, Daviet D, Savill J, Ren Y, Leung LLK, McGregor JL. Identification of a domain (155-183) on CD36 implicated in the phagocytosis of apoptotic neutrophils. J Biol Chem 1996; 271: 15381-5.
• 23 Morgan DML. Isolation and culture of human umbilical vein endothelial cells. In: GE Jones eds. Methods in Molecular Medicine: Human cell culture protocols. New Jersey. Pa: Humana Stress Inc.; 1995: 101-9.
• 24 Sobrevia L, Cesare P, Yudilevich DL, Mann GE. Activation of l-arginine transport (system y+) and nitric oxide synthase by elevated glucose and insulin in human endothelial cells. J Physiol 1996; 490: 775-81.
• 25 Wang AM, Doyle MV, Mark DF. Quantification of mRNA by the polymerase chain reaction. Proc Natl Acad Sci USA 1989; 86: 9717-21.
• 26 Kobayashi T, Vischer UM, Rosnoblet C, Lebrand C, Lindsay M, Parton RG, Kruithof EKO, Gruenberg J. The tetraspanin CD63/lamp3 cycles between endocytic and secretory compartments in human endothelial cells. Mol Biol Cell. 2000; 11: 1829-43.
• 27 Jilma B, Fasching P, Ruthner C, Rumplmayr A, Ruzicka S, Kapiotis S, Wagner OF, Eichler HG. Elevated circulating P-selectin in insulin-dependent diabetes mellitus. Thromb Haemost 1996; 76: 328-32.
• 28 Whelan J. Selectin synthesis and inflammation. TIBS 1996; 21: 65-9.
• 29 Vehkavaara S, Makimattila S, Schlenza A, Vakkilainen J, Westerbacka J, Yki-Jarvinen H. Insulin therapy improves endothelial function in type 2 diabetes. Art Thromb Vasc Biol 2000; 30: 500-45.
• 30 Nagel T, Resnick N, Atkinson WJ, Dewey CF, Gimbrone MA. Shear stress selectively upregulates intercellular adhesion molecule -1 expression in cultured human vascular endothelial cells. J Clin Inv 1994; 94: 885-91.
• 31 Ceriello A. Acute hyperglycaemia and oxidative stress generation. Diabetic Medicine. 1997; 14: 45-9.
• 32 Inoguchi T, Xia P, Kunisaki M, Higashi S, Feener EP, King GL. Insulin’s effect on protein kinase C and diacylglycerol induced by diabetes and glucose in vascular tissues. Am J Physiol (Endocrinol Metab). 1994; 30: 369-79.
• 33 Murohara T, Scalia R, Lefer M. Lysophosphatidylcholine promotes P-selectin expression in platelets and endothelial cells. Possible involvement of protein kinase C activation and its inhibition by nitric oxide donors. Circ Res 1996; 78: 780-9.
• 34 Takahashi M, Ikeda U, Masuyama J-I, Funayama H, Kano S, Shimada K. Nitric oxide attenuates adhesion molecule expression in human endothelial cells. Cytokine 1996; 8: 817-21.
• 35 Kashiwagi A, Asahina T, Nishio Y, Ikebuchi M, Tanaka Y, Kikkawa R, Shigeta Y. Glycation, oxidative stress, and scavenger activity: glucose metabolism and radical scavenger dysfunction in endothelial cells. Diabetes 1996; 45: 84-6.
• 36 Du X, Stocklauser-Farber Rosen P. Generation of reactive oxygen intermediates, activation of NF-KappaB and induction of apoptosis in human endothelial cells by glucose. Role of nitric oxide synthase? Free radical Biology & Medicine 1999; 27: 752-63.
• 37 Patel KD, Zimmerman GA, Prescott SM, McEver RP, McIntyre TM. Oxygen radicals induce human endothelial cells to express GMP-140 and bind neutrophils. J Cell Biol 1991; 112: 749-59.