caa a22 4500 475771745 CHVBK 20180406123619.0 cr unu---uuuuu 170329e20001101xx s 000 0 eng 10.1007/s001250051567 doi (NATIONALLICENCE)springer-10.1007/s001250051567 High glucose-induced intercellular adhesion molecule-1 (ICAM-1) expression through an osmotic effect in rat mesangial cells is PKC-NF-ϰB-dependent [Elektronische Daten] [C. W. Park, J. H. Kim, J. W. Lee, Y. S. Kim, H. J. Ahn, Y. S. Shin, S. Y. Kim, E. J. Choi, Y. S. Chang, B. K. Bang] Abstracts: Aims/hypothesis. Infiltration of mononuclear cells and glomerular enlargement accompanied by glomerular cell proliferation are very early characteristics of the pathophysiology of diabetes. To clarify the mechanism of early diabetic nephropathy, we measured [3H]-thymidine incorporation and cell numbers to show the influence of a high ambient glucose concentration and the osmotic effect on rat mesangial cell proliferation. We also measured the effect of high glucose on the expression of intercellular adhesion molecule-1 and vascular adhesion molecule-1 by flow cytometry and semiquantitative RT-PCR in mesangial cells and the adhesion of leukocytes to mesangial cells. Methods/results. Cells exposed to high d-glucose (30 mmol/l) caused an increase in [3H]-thymidine incorporation and cell numbers at 24 and 48 h and normalized at 72 h (p < 0.05), whereas these changes were not found in high mannitol (30 mmol/l), IL-1β, or TNFα-stimulated mesangial cells. Cells exposed to high-glucose (15, 30, or 60 mmol/l) or osmotic agents (l-glucose, raffinose and mannitol) showed that intercellular adhesion molecule-1 expression began to increase after 24 h, reached its maximum at 24 and 48 h and gradually decreased afterwards. The stimulatory effects of high glucose and high mannitol on mRNA expression were observed as early as 6 h and reached its maximum at 12 h. Up-regulation of ICAM-1 protein and mRNA was also found in IL-1-β and TNF-α-stimulated mesangial cells. Neither vascular adhesion molecule-1 protein nor mRNA expression was, however, affected by high glucose and high mannitol. Notably, the protein kinase C inhibitors calphostin C and staurosporine reduced high glucose- or high mannitol-induced intercellular adhesion molecule-1 mRNA expression and high glucose-induced proliferation. Furthermore, the NF-ϰB inhibitor N-tosyl-l-phenylalanine chloromethyl ketone reduced high glucose- or high mannitol-induced intercellular adhesion molecule-1 mRNA expression and high glucose-induced proliferation. Results showed that high glucose (15, 30 mmol/l) or high concentrations of osmotic agents remarkably increased the number of adherent leukocytes to mesangial cells (p < 0.01) compared with control cells (5 mmol/l d-glucose). Functional blocking of intercellular adhesion molecule-1 on mesangial cells with rat intercellular adhesion molecule-1 monoclonal antibody, calphostin C, staurosporine, or N-tosyl-l-phenylalanine chloromethyl ketone significantly inhibited high glucose- or high mannitol-induced increase in leukocyte adhesion (p < < 0.05). Conclusion/interpretation. These results suggest that high glucose can upregulate intercellular adhesion molecule-1 protein and mRNA expression but not vascular adhesion molecule-1 expression in mesangial cells and promote leukocyte adhesion through up-regulation of intercellular adhesion molecule-1 through osmotic effect, possibly depending on the protein kinase C nuclear factor-kappa B (PKC-NF-ϰB) pathway. High glucose itself can also promote mesangial cell proliferation through the PKC-NF-ϰB pathways. We conclude that hyperglycaemia in itself seems to be an important factor in the development of early diabetic nephropathy. [Diabetologia (2000) 43: 1544-1553] Springer-Verlag Berlin Heidelberg, 2000 Keywords Diabetes mellitus nationallicence glucose nationallicence ICAM-1 nationallicence mesangial cells nationallicence PKC-NF-ϰB nationallicence Park C. W. Division of Nephrology, Department of Internal Medicine, XX aut Kim J. H. Department of Obstetrics and Gynecology, The Catholic University of Korea, Seoul, Korea, KR aut Lee J. W. Department of Biochemistry, The Catholic University of Korea, Seoul, Korea, KR aut Kim Y. S. Division of Nephrology, Department of Internal Medicine, XX aut Ahn H. J. Division of Nephrology, Department of Internal Medicine, XX aut Shin Y. S. Division of Nephrology, Department of Internal Medicine, XX aut Kim S. Y. Division of Nephrology, Department of Internal Medicine, XX aut Choi E. J. Division of Nephrology, Department of Internal Medicine, XX aut Chang Y. S. Division of Nephrology, Department of Internal Medicine, XX aut Bang B. K. Division of Nephrology, Department of Internal Medicine, XX aut https://doi.org/10.1007/s001250051567 text/html Onlinezugriff via DOI 1 research-article jats NATIONALLICENCE 856 40 https://doi.org/10.1007/s001250051567 text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- Park C. W. Division of Nephrology, Department of Internal Medicine, XX aut NATIONALLICENCE 700 1- Kim J. H. Department of Obstetrics and Gynecology, The Catholic University of Korea, Seoul, Korea, KR aut NATIONALLICENCE 700 1- Lee J. W. Department of Biochemistry, The Catholic University of Korea, Seoul, Korea, KR aut NATIONALLICENCE 700 1- Kim Y. S. Division of Nephrology, Department of Internal Medicine, XX aut NATIONALLICENCE 700 1- Ahn H. J. Division of Nephrology, Department of Internal Medicine, XX aut NATIONALLICENCE 700 1- Shin Y. S. Division of Nephrology, Department of Internal Medicine, XX aut NATIONALLICENCE 700 1- Kim S. Y. Division of Nephrology, Department of Internal Medicine, XX aut NATIONALLICENCE 700 1- Choi E. J. Division of Nephrology, Department of Internal Medicine, XX aut NATIONALLICENCE 700 1- Chang Y. S. Division of Nephrology, Department of Internal Medicine, XX aut NATIONALLICENCE 700 1- Bang B. K. Division of Nephrology, Department of Internal Medicine, XX aut Metadata rights reserved Springer special CC-BY-NC licence nationallicence BK010053 XK010053 XK010000 NATIONALLICENCE NATIONALLICENCE NL-springer