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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)

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Abstract

An increase in the interaction between advanced glycation end-products (AGEs) and their receptor RAGE is believed to contribute to the pathogenesis of chronic complications of Diabetes mellitus, which can include bone alterations such as osteopenia. We have recently found that extracellular AGEs can directly regulate the growth and development of rat osteosarcoma UMR106 cells, and of mouse calvaria-derived MC3T3E1 osteoblasts throughout their successive developmental stages (proliferation, differentiation and mineralisation), possibly by the recognition of AGEs moieties by specific osteoblastic receptors which are present in both cell lines. In the present study we examined the possible expression of RAGE by UMR106 and MC3T3E1 osteoblastic cells, by immunoblot analysis. We also investigated whether short-, medium- or long-term exposure of osteoblasts to extracellular AGEs, could modify their affinity constant and maximal binding for AGEs (by 125I-AGE-BSA binding experiments), their expression of RAGE (by immunoblot analysis) and the activation status of the osteoblastic ERK 1/2 signal transduction mechanism (by immunoblot analysis for ERK and P-ERK). Our results show that both osteoblastic cell lines express readily detectable levels of RAGE. Short-term exposure of phenotypically mature osteoblastic UMR106 cells to AGEs decrease the cellular density of AGE-binding sites while increasing the affinity of these sites for AGEs. This culture condition also dose-dependently increased the expression of RAGE and the activation of ERK. In proliferating MC3T3E1 pre-osteoblasts, 24–72 h exposure to AGEs did not modify expression of RAGE, ERK activation or the cellular density of AGE-binding sites. However, it did change the affinity of these binding sites for AGEs, with both higher- and lower-affinity sites now being apparent. Medium-term (1 week) incubation of differentiated MC3T3E1 osteoblasts with AGEs, induced a simultaneous increase in RAGE expression and in the relative amount of P-ERK. Mineralising MC3T3E1 cultures grown for 3 weeks in the presence of extracellular AGEs showed a decrease both in RAGE and P-ERK expression. These results indicate that, in phenotypically mature osteoblastic cells, changes in ERK activation closely follow the AGEs-induced regulation of RAGE expression. Thus, the AGEs-induced biological effects that we have observed previously in osteoblasts, could be mediated by RAGE in the later stages of development, and mediated by other AGE receptors in the earlier pre-osteoblastic stage.

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References

  1. Vlassara H, Bucala R, Striker L: Pathogenic effects of advanced glycosylation: Biochemical, biologic and clinical implications for diabetes and ageing. Lab Invest 70: 138-151, 1994

    Google Scholar 

  2. Baynes JW, Thorpe SR: Role of oxidative stress in diabetic complications: A new perspective on an old paradigm. Diabetes 48: 1-9, 1999

    Google Scholar 

  3. Thornalley PJ: Cell activation by glycated proteins. Age receptors, receptor recognition factors and functional classification of AGEs. Cell Mol Biol 44: 1013-1033, 1998

    Google Scholar 

  4. Araki N, Higashi T, Mori T, Shibayama R, Kawama T, Takahashi K, Shichiri M, Horiuchi S: Macrophage scavenger receptor mediates the endocytic uptake and degradation of advanced glycation and products of the Maillard reaction. Eur J Biochem 230: 408-415, 1995

    Google Scholar 

  5. Suzuki H, Kurihara Y, Takeya M, Kamada N, Kataoka M, Jishage K, Ueda O, Sakaguchi H, Higashi T, Suzuki T et al.: A role for macrophage scavenger receptors in atherosclerosis and susceptibility to infection. Nature (London) 386: 292, 296, 1997

    Google Scholar 

  6. Miyata T, Kawai R, Taketomi S, Sprague SM: Possible involvement of advanced glycation end-products in bone resorption. Nephrol Dial Transpl 11: 54-57, 1996

    Google Scholar 

  7. 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 11: 931-937, 1996

    Google Scholar 

  8. Takagi M, Kasayama S, Yamamoto T, Motomura T, Hashimoto K, Yamamoto H, Sato B, Okada S, Kishimoto T: Advanced glycation end-products stimulate interleukin-6 production by human bone-derived cells. J Bone Miner Res 12: 439-446, 1997

    Google Scholar 

  9. McCarthy AD, Etcheverry SB, Bruzzone L, Cortizo AM: Effects of advanced glycation end-products on the proliferation and differentiation of osteoblast-like cells. Mol Cell Biochem 170: 43-51, 1997

    Google Scholar 

  10. McCarthy AD, Etcheverry SB, Cortizo AM: Effect of advanced glycation end-products on the secretion of insulin-like growth factor-I and its binding proteins: Role in osteoblast development. Acta Diabetol 38: 113-122, 2001

    Google Scholar 

  11. Celic S, Katayama Y, Chilco PJ, Martin TJ, Findlay DM: Type I collagen influence on gene expression in UMR106-06 osteoblast-like cells is inhibited by genistein. J Endocrinol 158: 377-388, 1998

    Google Scholar 

  12. Paul RG, Bailey AJ: The effect of advanced glycation end-product formation upon cell-matrix interactions. Int J Biochem Cell Biol 31: 653-660, 1999

    Google Scholar 

  13. McCarthy AD, Etcheverry SB, Cortizo AM: Advanced glycation end-product-specific receptors in rat and mouse osteoblast-like cells: Regulation with stages of differentiation. Acta Diabetol 36: 45-52, 1999

    Google Scholar 

  14. McCarthy AD, Etcheverry SB, Bruzzone L, Lettieri MG, Barrio DA, Cortizo AD: Non-enzymatic glycosylation of a type I collagen matrix: Effects on osteoblastic development and oxidative stress. BMC Cell Biol 2: 16, 2001

    Google Scholar 

  15. Kawano E, Takahashi S-I, Sakano Y, Fujimoto D: Non enzymatic glycosylation alters properties of collagen as a substratum for cells. Matrix 10: 300-305, 1990

    Google Scholar 

  16. Partridge NC, Alcorn D, Michelangeli VP, Ryan G, Martin TJ: Morphological and biochemical characterization of four clonal osteogenic sarcoma cell lines of rat origin. Cancer Res: 43: 4308-4312, 1983

    Google Scholar 

  17. Takeuchi Y, Nakayama K, Matsumoto T: Differentiation and cell surface expression of transforming growth factor-β receptors are regulated by interaction with matrix collagen in murine osteoblastic cells. J Biol Chem 271: 3938-3944, 1996

    Google Scholar 

  18. Quarles LD, Yahay DA, Lever LW, Caton R, Wenstrup RJ: Distinct proliferative and differentiated stages of murine MC3T3E1 cells in culture: An in vitro model of osteoblast development. J Bone Miner Res 7: 683-692, 1992

    Google Scholar 

  19. Laemmli UK: Cleavage of structural protein during the assembly of the head of bacteriophage T4. Nature 227: 680-685, 1970

    Google Scholar 

  20. Lowry OH, Rosebrough NJ, Farr AJ, Randall RJ: Protein measurement with Folin phenol reagent. J Biol Chem 193: 265-275, 1951

    Google Scholar 

  21. Lue L-F, Walker DG, Brachova L, Beach TG, Rogers J, Schmidt AM, Stern DM, Yan SD: Involvement of microglial receptor for advanced glycation end-products (RAGE) in Alzheimer's disease: Identification of a cellular activation mechanism. Exp Neurol 171: 29-45, 2001

    Google Scholar 

  22. Schmidt AM, Stern DM: RAGE: A new target for the prevention and treatment of the vascular and inflammatory complications of diabetes. TEM 11: 368-375, 2000

    Google Scholar 

  23. Lander HM, Tauras JM, Ogiste JS, Hori O, Moss RA, Schmidt AM: Activation of the receptor for advanced glycation end-products triggers a p21ras-dependent-mitogen-activated protein kinase pathway regulated by oxidant stress. J Biol Chem 272: 17810-17814, 1997

    Google Scholar 

  24. Huttunen HJ, Fages C, Rauvala H: Receptor for advanced glycation end-products (RAGE)-mediated neurite outgrowth and activation of NF-κB require the cytoplasmic domain of the receptor but different downstream signalling pathways. J Biol Chem 274: 19919-19924, 1999

    Google Scholar 

  25. Li J, Schmidt AM: Characterization and functional analysis of the promoter of RAGE, the receptor for advanced glycation end-products. J Biol Chem 272: 16498-16506, 1997

    Google Scholar 

  26. Bierhaus A, Schiekofer S, Schwaninger M, Andrassy M, Humpert PM, Chen J, Hong M, Luther T, Henle T, Klöting I, Morcos M, Hofmann M, Tritschler H, Weigle B, Kasper M, Smith M, Perry G, Schmidt AM, Stern DM, Häring H-U, Schleicher E, Nawroth PP: Diabetes-associated sustained activation of the transcription factor nuclear factor-κB. Diabetes 50: 2792-2808, 2001

    Google Scholar 

  27. Beg AA, Baltimore D: An essential role of NF-κB in preventing TNFα-induced cell death. Science 274: 782-784, 1996

    Google Scholar 

  28. Tomasek JJ, Meyers SW, Basinger JB, Green DJ, Shew RL: Diabetic and age-related enhancement of collagen-linked fluorescence in cortical bones of rats. Life Sci 55: 855-861, 1994

    Google Scholar 

  29. Krakauer JC, McKenna MJ, Buderer NF, Rao DS, Whitehouse FW, Parfitt AM: Bone loss and bone turnover in diabetes mellitus. Diabetes 44: 775-782, 1995

    Google Scholar 

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Cortizo, A.M., Lettieri, M.G., Barrio, D.A. et al. 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 250, 1–10 (2003). https://doi.org/10.1023/A:1024934008982

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