Calcified Tissue International

, Volume 52, Issue 1, pp 62–66 | Cite as

Protein kinase C mediates flow-induced prostaglandin E2 production in osteoblasts

  • Kathleen M. Reich
  • John A. Frangos
Laboratory Investigations

Summary

Interstitial fluid flow generated by skeletal loading may be responsible for load-induced bone remodeling. Production of prostaglandin E2 (PGE2), a potent mediator of bone remodeling, is augmented in osteoblasts exposed to fluid flow. Exposure to fluid flow resulted in a slight initial increase in PGE2 production (1–2 hour), followed by a dramatic increase (2–8 hours). The initial phase of only slightly increased PGE2 production was dependent on substrate availability. H7, a protein kinase C inhibitor, strongly inhibited flow-induced prostaglandin E2 production at all time points examined without effecting production in stationary cultures. Blocking protein synthesis with cycloheximide resulted in a 56% reduction in long-term flow-induced PGE2 production. Thus, the later phase appeared to be the result of an increased number of enzymes as well as increased activity of existing enzymes or increased substrate availability. In conclusion, fluid flow increases PGE2 production in osteoplasts via a protein kinase C-dependent pathway involvingde novo protein synthesis.

Key words

Signal transduction Bone remodeling Shear stress 

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References

  1. 1.
    Smith EL, Gilligan C (1989) Mechanical forces and bone. Bone Miner Res 6:139–173Google Scholar
  2. 2.
    Frost HM (1990) Skeletal structural adaptation to mechanical usage (SATMU) 1. Redefining Wolff's law: the bone remodelling problem. Anat Rec 226:403–413Google Scholar
  3. 3.
    Okuda A, Taylor LM, Heersche JNM (1989) Prostaglandin E2 initially inhibits and then stimulates bone resorption in isolated rabbit osteoclast cultures. Bone Miner 7:255–266Google Scholar
  4. 4.
    Jee WSS, Ueno K, Kimmel DB, Woodbury DM, Price P, Woodbury LA (1987) The role of bone cells in increasing metaphyseal hard tissue in rapidly growing rats treated with prostaglandin E2. Bone 8:171–178Google Scholar
  5. 5.
    Reich KM, Frangos JA (1991) Effect of flow on prostaglandin E2 production and inositol trisphosphate levels in osteoblasts. Am J Physiol 261:C428-C432Google Scholar
  6. 6.
    Harell A, Dekel S, Binderman I (1977) Biochemical effect of mechanical stress on cultured bone cells. Calcif Tissue Int 22: 202–209Google Scholar
  7. 7.
    Somjen D, Binderman I, Berger E, Harell A (1980) Bone remodelling induced by physical stress is prostaglandin E2 mediated. Biochim Biophys Acta 627:91–100Google Scholar
  8. 8.
    Yeh C, Rodan GA (1984) Tensile forces enhance prostaglandin E2 synthesis in osteoblastic cells grown on collagen ribbons. Calcif Tissue Int 36:S67-S71Google Scholar
  9. 9.
    Pead MJ, Lanyon LE (1989) Indomethacin modulation of load-related stimulation of new bone formation in vivo. Calcif Tissue Int 45:34–40Google Scholar
  10. 10.
    Ogita K, Koide H, Kikkawa U, Kishimoto A, Nishizuka Y (1990) The heterogeneity of protein kinase C in signal transduction cascade. In: Nishizuka Y (eds) The biology and medicine of signal transduction. Raven Press, New York, p 218Google Scholar
  11. 11.
    Hsieh H, Li N, Frangos JA (1992) Shear-induced plateletderived growth factor gene expression in human endothelial cells is mediated by protein kinase C. J Cell Physiol 150:552–558Google Scholar
  12. 12.
    Ecarot-Charrier B, Glorieux H, Van der Rest M, Pereira G (1983) osteoblasts isolated from mouse calvaria initiate matrix mineralization in culture. J Cell Biol 96:639–643Google Scholar
  13. 13.
    Reich KM, Gay CV, Frangos JA (1990) Fluid shear stress as a mediator of cyclic adenosine monophosphate production. J Cell Physiol 143:100–104Google Scholar
  14. 14.
    Frangos JA, Eskin SG, McIntire LV (1988) Shear stress induced mammalian cell stimulation. Biotech Bioeng 32:1053–1060Google Scholar
  15. 15.
    Hidaka H, Inagaki M, Kawamoto S, Sasaki Y (1984) Isoquinolinesulfonamides, novel and potent inhibitors of cyclic nucleotide-dependent protein kinase and protein kinase C. Biochemistry 23:5036–5041Google Scholar
  16. 16.
    Neter J, Wasserman W, Kutner MH (1985) Applied linear statistical models. Richard D. Irwin, Inc., Homewood, ILGoogle Scholar
  17. 17.
    Carter TD, Hallam TJ, Pearson JD (1989) Protein kinase C activation alters the sensitivity of agonist-stimulated endothelial cell prostacyclin production to intracellular Ca2+ Biochem J 262:431–437Google Scholar
  18. 18.
    Sander J, Myatt L (1990) Regulation of prostaglandin E2 synthesis in human amnion by protein kinase C. Prostaglandins 39:355–363Google Scholar
  19. 19.
    Aizu E, Yamamoto S, Nakadate T, Kato R (1990) Differential effects of various skin tumor-promoting agents on prostaglandin E2 release from primary cultures of mouse epidermal cells. Eur J Pharmacol 182:19–28Google Scholar
  20. 20.
    Hagel-Bradway S, Dziak R (1989) Regulation of bone cell metabolism. J Oral Pathol Med 18:344–351Google Scholar
  21. 21.
    Ikeda K, Sugimoto T, Fukase M, Fujita T (1991) Protein kinase C is involved in PTH-induced homologous desensitization by directly affecting PTH receptor in the osteoblastic osteosarcoma cells. Endocrinology 128:2901–2906Google Scholar
  22. 22.
    Kawase T, Suzuki A (1990) Initial responses of a clonal osteoblast-like cell line, MOB 3–4, to phosphatidic acid in vitro. Bone Miner 10:61–70Google Scholar
  23. 23.
    Hagel-Bradway S, Tatakis DN, Dziak R (1991) Prostaglandin-induced changes in calcium uptake and cAMP production in osteoblast-like cells: role of protein kinase C. Calcif Tissue Int 48:272–277Google Scholar
  24. 24.
    Klein-Nulend J, Pilbeam CC, Harrison JR, Fall PM, Raisz LG (1991) Mechanism of regulation of prostaglandin production by parathyroid hormone, interleukin-1, and cortisol in cultured mouse parietal bones. Endocrinology 128:2503–2510Google Scholar
  25. 25.
    Sumitani K, Kawata T, Yoshimoto T, Yamamoto S, Kumegawa M (1989) Fatty acid cyclooxygenase activity stimulated by transforming growth factor-β in mouse osteoblastic cells (MC3T3-E1). Arch Biochem Biophys 270:588–595Google Scholar
  26. 26.
    Yokota K, Kusaka M, Ohshima T, Yamamoto S, Kurihari N, Yoshino T, Kumegawa M (1986) Stimulation of prostaglandin E2 synthesis in cloned osteoblastic cells of mouse (MC3T3-E1) by epidermal growth factor. J Biol Chem 261:15410–15415Google Scholar
  27. 27.
    Rubin CT, Lanyon LE (1984) Regulation of bone formation by applied dynamic loads. J Bone Joint Surg 66A:397–402Google Scholar

Copyright information

© Springer-Verlag New York Inc. 1993

Authors and Affiliations

  • Kathleen M. Reich
    • 1
  • John A. Frangos
    • 1
  1. 1.Department of Chemical Engineering, 150 Fenske LaboratoryThe Pennsylvania State UniversityUniversity ParkUSA

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