The Combination of High Glucose and Advanced Glycation End-products (AGEs) Inhibits the Mineralization of Osteoblastic MC3T3-E1 Cells through Glucose-induced Increase in the Receptor for AGEs

Abstract

Type 1 diabetes mellitus is known to be associated with reduced bone mass and increased bone fractures. This is thought to be due to a decrease in osteoblastic bone formation rather than an increase in osteoclastic bone resorption, but the precise mechanism is unknown. In this study, we examined whether or not high glucose or advanced glycation end-products (AGEs), which play key roles in the pathogenesis and complications of diabetes, affect the differentiation of osteoblastic MC3T3-E1 cells. First, MC3T3-E1 cells were incubated in media containing either 22 mM glucose, 22 mM mannitol, 300 μg/ml AGE2, or 300 μg/ml AGE3. Each of these agents alone did not affect the mineralization of the cells by von Kossa staining and Alizarin red staining. However, high glucose but not mannitol or AGEs markedly increased mRNA expression of AGE receptor (RAGE) by real-time PCR. Next, we examined the combined effects of high glucose and AGEs on the differentiation of MC3T3-E1 cells. The combination of 22 mM glucose and 300 μg/ml AGE2 significantly inhibited the mineralization of MC3T3-E1 cells, and 22 mM glucose in combination with either 300 μg/ml AGE2 or AGE3 apparently decreased osteocalcin mRNA expression. These results suggest that high glucose or AGEs alone might have no effect on osteoblastic differentiation, but their combination could additionally or synergistically inhibit osteoblastic mineralization through glucose-induced increase in RAGE expression.

Key words

diabetes mellitus - osteoblast - bone formation - differentiation - RAGE

References

• 1 Forsen L, Meyer HE, Midthjell K, Edna TH. Diabetes mellitus and the incidence of hip fracture: results from the Nord-Trondelag Health Survey.  Diabetologia. 1999;  42 920-925
• 2 Nicodemus KK, Folsom AR. Type 1 and type 2 diabetes and incident hip fractures in postmenopausal women.  Diabetes Care. 2001;  24 1192-1197
• 3 Raskin P, Stevenson MR, Barilla DE, Pak CY. The hypercalciuria of diabetes mellitus: its amelioration with insulin.  Clin Endocrinol (Oxf). 1978;  9 329-335
• 4 Ishida H, Cunningham NS, Henry HL, Norman AW. The number of 1,25-dihydroxyvitamin D3 receptors is decreased in both intestine and kidney of genetically diabetic db/db mice.  Endocrinology. 1988;  122 2436-2443
• 5 Sugimoto T, Ritter C, Morrissey J, Hayes C, Slatopolsky E. Effects of high concentrations of glucose on PTH secretion in parathyroid cells.  Kidney Int. 1990;  37 1522-1527
• 6 Jehle PM, Jehle DR, Mohan S, Bohm BO. Serum levels of insulin-like growth factor system components and relationship to bone metabolism in Type 1 and Type 2 diabetes mellitus patients.  J Endocrinol. 1998;  159 297-306
• 7 Bouillon R, Bex M, Van Herck E, Laureys J, Dooms L, Lesaffre E, Ravussin E. Influence of age, sex, and insulin on osteoblast function: osteoblast dysfunction in diabetes mellitus.  J Clin Endocrinol Metab. 1995;  80 1194-1202
• 8 Botolin S, Faugere MC, Malluche H, Orth M, Meyer R, MacCabe LR. Increased bone adiposity and peroxisomal proliferator-activated receptor-gamma2 expression in type I diabetic mice.  Endocrinology. 2005;  146 3622-3631
• 9 Brenner RE, Riemenschneider B, Blum W, Morike M, Teller WM, Pirsig W, Heinze E. Defective stimulation of proliferation and collagen biosynthesis of human bone cells by serum from diabetic patients.  Acta Endocrinol (Copenh). 1992;  127 509-514
• 10 Terada M, Inaba M, Yano Y, Hasuma T, Nishizawa Y, Morii H, Otani S. Growth-inhibitory effect of a high glucose concentration on osteoblast-like cells.  Bone. 1998;  22 17-23
• 11 Botolin S, MacCabe LR. Chronic hyperglycemia modulates osteoblast gene expression through osmotic and non-osmotic pathways.  J Cell Biochem. 2006;  99 411-424
• 12 Gopalakrishnan V, Vignesh RC, Arunakaran J, Aruldhas MM, Srinivasan N. Effects of glucose and its modulation by insulin and estradiol on BMSC differentiation into osteoblastic lineages.  Biochem Cell Biol. 2006;  84 93-101
• 13 Yamagishi S, Amano S, Inagaki Y, Okamoto T, Koga K, Sasaki N, Yamamoto H, Takeuchi M, Makita Z. Advanced glycation end products-induced apoptosis and overexpression of vascular endothelial growth factor in bovine retinal pericytes.  Biochem Biophys Res Commun. 2002;  290 973-978
• 14 Yamagishi S, Inagaki Y, Okamoto T, Amano S, Koga K, Takeuchi M, Makita Z. Advanced glycation end product-induced apoptosis and overexpression of vascular endothelial growth factor and monocyte chemoattractant protein-1 in human-cultured mesangial cells.  J Biol Chem. 2002;  277 20309-20315
• 15 Sekido H, Suzuki T, Jomori T, Takeuchi M, Yabe-Nishimura C, Yagihashi S. Reduced cell replication and induction of apoptosis by advanced glycation end products in rat Schwann cells.  Biochem Biophys Res Commun. 2004;  320 241-248
• 16 Fukami K, Ueda S, Yamagishi S, Kato S, Inagaki Y, Takeuchi M, Motomiya Y, Bucala R, Iida S, Tamaki K, Imaizumi T, Cooper ME, Okuda S. AGEs activate mesangial TGF-beta-Smad signaling via an angiotensin II type I receptor interaction.  Kidney Int. 2004;  66 2137-2147
• 17 Takagi M, Kasayama S, Yamamoto T, Motomura T, Hashimoto K, Yamamoto H, Sato B, Okada S, Kishimoto T. Advanced glycation endproducts stimulate interleukin-6 production by human bone-derived cells.  J Bone Miner Res. 1997;  12 439-446
• 18 Yamamoto T, Ozono K, Miyauchi A, Kasayama S, Kojima Y, Shima M, Okada S. Role of advanced glycation end products in adynamic bone disease in patients with diabetic nephropathy.  Am J Kidney Dis. 2001;  38 S161-S164
• 19 Katayama Y, Akatsu T, Yamamoto M, Kugai N, Nagata N. Role of nonenzymatic glycosylation of type I collagen in diabetic osteopenia.  J Bone Miner Res. 1996;  11 931-937
• 20 Sudo H, Kodama HA, Amagai Y, Yamamoto S, Kasai S. In vitro differentiation and calcification in a new clonal osteogenic cell line derived from newborn mouse calvaria.  J Cell Biol. 1983;  96 191-198
• 21 Kume S, Kato S, Yamagishi S, Inagaki Y, Ueda S, Arima N, Okawa T, Kojiro M, Nagata K. Advanced glycation end-products attenuate human mesenchymal stem cells and prevent cognate differentiation into adipose tissue, cartilage, and bone.  J Bone Miner Res. 2005;  20 1647-1658
• 22 Alikhani M, Alikhani Z, Boyd C, Maclellan CM, Raptis M, Liu R, Pischon N, Trackman PC, Gerstenfeld L, Graves DT. Advanced glycation end products stimulate osteoblast apoptosis via the MAP kinase and cytosolic apoptotic pathways.  Bone. 2007;  40 345-353
• 23 Cortizo AM, Lettieri MG, Barrio DA, Mercer N, Etcheverry SB, MacCarthy AD. Advanced glycation end-products (AGEs) induce concerted changes in the osteoblastic expression of their receptor RAGE and in the activation of extracellular signal-regulated kinases (ERK).  Mol Cell Biochem. 2003;  250 1-10
• 24 Hatakeyama D, Kozawa O, Niwa M, Matsuno H, Ito H, Kato K, Tatematsu N, Shibata T, Uematsu T. Upregulation by retinoic acid of transforming growth factor-beta-stimulated heat shock protein 27 induction in osteoblasts: involvement of mitogen-activated protein kinases.  Biochim Biophys Acta. 2002;  1589 15-30
• 25 Tokuda H, Hatakeyama D, Akamatsu S, Tanabe K, Yoshida M, Shibata T, Kozawa O. Involvement of MAP kinases in TGF-beta-stimulated vascular endothelial growth factor synthesis in osteoblasts.  Arch Biochem Biophys. 2003;  415 117-125
• 26 Kanno Y, Ishisaki A, Yoshida M, Tokuda H, Numata O, Kozawa O. SAPK/JNK plays a role in transforming growth factor-beta-induced VEGF synthesis in osteoblasts.  Horm Metab Res. 2005;  37 140-145
• 27 Noda T, Tokuda H, Yoshida M, Yasuda E, Hanai Y, Takai S, Kozawa O. Possible involvement of phosphatidylinositol 3-kinase/Akt pathway in insulin-like growth factor-I-induced alkaline phosphatase activity in osteoblasts.  Horm Metab Res. 2005;  37 270-274
• 28 Takai S, Tokuda H, Hanai Y, Kozawa O. Phosphatidylinositol 3-kinase/akt plays a part in tumor necrosis factor-alpha-induced interleukin-6 synthesis in osteoblasts.  Horm Metab Res. 2006;  38 563-569

Correspondence