Skip to main content

Induction of heme oxygenase-1 and heat shock protein 70 in rat hepatocytes: The role of calcium signaling


Stress response genes including heat shock proteins are induced under a variety of conditions to confer cellular protection. This study investigated the role of calcium signaling in the induction of two stress response genes, heme oxygenase-1/hsp32 and hsp70, in isolated rat hepatocytes. Both genes were induced by cellular glutathione depletion. This induction could be inhibited by BAPTA-AM. Culturing in a calcium-free medium prevented the induction of hsp70 gene expression after glutathione depletion without affecting heme oxygenase-1 gene expression. Thapsigargin increased the gene expression of heme oxygenase-1 but not that of hsp70. Thapsigargin-induced heme oxygenase-1 induction was completely inhibited by BAPTA-AM. Incubation with the Ca2+-ionophore A23187 augmented heme oxygenase-1 (two-fold) and hsp70 (5.2-fold) mRNA levels. Our data suggests a significant role of Ca2+-dependent pathways in the induction of the two stress genes. An increase in the cytoplasmic Ca2+ activity seems to play a key role in the cascade of signaling leading to the induction of the two genes. However, the source of Ca2+ that fluxes into the cytoplasm seems to be different. Our data provides evidence for a compartmentalization of calcium fluxes, i.e. the Ca2+ flux from intracellular stores (e.g. the endoplasmic reticulum) plays a major role in the induction of heme oxygenase-1. By contrast, Ca2+ flux from the extracellular medium seems to be a mechanism initiating the cellular signaling cascade leading to hsp70 gene induction.



activator protein-1


(1,2-bis(o-amino-phenoxy)ethane-N,N,N′,N′-tetra (acetoxymethyl)esther






endoplasmic reticulum




heme oxygenase-1


heat shock protein


Krebs-Henseleit buffer


phosphate buffered saline


2,6 dimethyl-2,5 heptadien-4-on (phorone)/buthionine sulfoximine




  1. Beck, S.C., Paidas, C.N., Mooney, M., Deitch, E.A. and De Maio, A. Presence of the stress-inducible form of hsp-70 (hsp-72) in normal rat colon. Shock 3 (1995) 398–402.

    PubMed  Article  CAS  Google Scholar 

  2. Bauer, I., Wanner, G.A., Rensing, H., Alte, C., Miescher, E.A., Wolf, B., Pannen, B.H., Clemens, M.G. and Bauer, M. Expression pattern of heme oxygenase isoenzymes 1 and 2 in normal and stress-exposed rat liver. Hepatology 27 (1998) 829–838.

    PubMed  Article  CAS  Google Scholar 

  3. Kiang, J.G. and Tsokos, G.C. Heat shock protein 70 kDa: molecular biology, biochemistry, and physiology. Pharmacol. Ther. 80 (1998) 183–201.

    PubMed  Article  CAS  Google Scholar 

  4. Rensing, H., Bauer, I., Peters, I., Wein, T., Silomon, M., Jaeschke, H. and Bauer, M. Role of reactive oxygen species for hepatocellular injury and heme oxygenase-1 expression following hemorrhage and resuscitation. Shock 12 (1999) 300–308.

    PubMed  Article  CAS  Google Scholar 

  5. Doi, Y., Hamazaki, K., Yabuki, M., Tanaka, N. and Utsumi, K. Effect of HSP70 induced by warm ischemia to the liver on liver function after partial hepatectomy. Hepatogastroenterology 48 (2001) 533–540.

    PubMed  CAS  Google Scholar 

  6. Terry, C.M., Clikeman, J.A., Hoidal, J.R. and Callahan, K.S. TNF-α and IL-1a induce heme oxygenase-1 via protein kinase C, Ca2+, and phospholipase A2 in endothelial cells. Am. J. Physiol. 276 (1999) H1493–H1501.

    PubMed  CAS  Google Scholar 

  7. Kiang, J.G., Kiang, S.C., Juang, Y-T. and Tsokos, G.C. Nw-nitro-L-arginine inhibits inducible HSP-70 via Ca2+, PKC, and PKA in human intestinal epithelial cells. Am. J. Physiol. 282 (2002) G415–G423.

    CAS  Google Scholar 

  8. Hardingham, G.E. and Bading, H. Calcium as a versatile second messenger in the control of gene expression. Microsc. Res. Tech. 46 (1999) 348–355.

    PubMed  Article  CAS  Google Scholar 

  9. Choi, A.M., Tucker, R.W., Carlson, S.G., Weigand, G. and Holbrook, N.J. Calcium mediates expression of stress-response genes in prostaglandin A2-induced growth arrest. FASEB J. 8 (1994) 1048–1054.

    PubMed  CAS  Google Scholar 

  10. Linden, T., Doutheil, J. and Paschen, W. Role of calcium in the activation of erp72 and heme oxygenase-1 expression on depletion of endoplasmic reticulum calcium stores in rat neuronal cell culture. Neurosci. Lett. 247 (1998) 103–106.

    PubMed  Article  CAS  Google Scholar 

  11. Koleva, M. and Stoytchev, T. On the enzyme-inducing action of calcium antagonists. Arch. Toxicol. 67 (1993) 294–296.

    PubMed  Article  CAS  Google Scholar 

  12. Elo, M.A., Sironen, R.K., Kaarniranta, K., Auriola, S., Helminen, H.J. and Lammi, M.J. Differential regulation of stress proteins by high hydrostatic pressure, heat shock, and unbalanced calcium homeostasis in chondrocytic cells. J. Cell. Biochem. 79 (2000) 610–619.

    PubMed  Article  CAS  Google Scholar 

  13. Sreedhar, A.S. and Srivinas, U.K. Activation of stress response by ionomycin in rat hepatoma cells. J. Cell. Biochem. 86 (2002) 154–161.

    PubMed  Article  CAS  Google Scholar 

  14. Rensing, H., Bauer, I., Datene, V., Patau, C., Pannen, B.H. and Bauer, M. Differential expression pattern of heme oxygenase-1/heat shock protein 32 and nitric oxide synthase-II and their impact on liver injury in a rat model of hemorrhage and resuscitation. Crit. Care Med. 27 (1999) 2766–2775.

    PubMed  Article  CAS  Google Scholar 

  15. Rensing, H., Jaeschke, H., Bauer, I., Pätau, C., Datene, V., Pannen, B.H. and Bauer, M. Differential activation pattern of redox-sensitive transcription factors and stress-inducible dilator systemsheme oxygenase-1 and inducible nitric oxide synthase in hemorrhagic and endotoxic shock. Crit. Care Med. 29 (2001) 1962–1971.

    PubMed  Article  CAS  Google Scholar 

  16. Rose, S., Pizanis, A. and Silomon, M. Altered hepato-cellular Ca2+ regulation during hemorrhagic shock and resuscitation. Hepatology 25 (1997) 379–384.

    PubMed  Article  CAS  Google Scholar 

  17. Strom, S.C. and Michalopoulos, G. Collagen as a substrate for cell growth and differentiation. Methods Enzymol. 82 (1982) 544–555.

    PubMed  CAS  Article  Google Scholar 

  18. Snyder, J.W., Serroni, A., Savory, J. and Farber, J.L. The absence of extracellular calcium potentiates the killing of cultured hepatocytes by aluminum maltolate. Arch. Biochem. Biophys. 316 (1995) 434–442.

    PubMed  Article  CAS  Google Scholar 

  19. Bauer, I., Rensing, H., Florax, A., Ulrich, C., Pistorius, G., Redl, H. and Bauer, M. Expression pattern and regulation of heme oxygenase-1/heat shock protein 32 in human liver cells. Shock 20 (2003) 116–222.

    PubMed  Article  CAS  Google Scholar 

  20. Mellstrom, B. and Naranjo, J.R. Mechanisms of Ca2+-dependent transcription. Curr. Opin. Neurobiol. 11 (2001) 312–319.

    PubMed  Article  CAS  Google Scholar 

  21. Thastrup, O., Cullen, P.J., Drobak, B.K., Hanley, M.R. and Dawson, A.P. Thapsigargin, a tumor promotor, discharges intracellular Ca2+ stores by specific inhibition of the endoplasmic reticulum Ca2+ ATPase. Proc. Natl. Acad. Sci. USA 87 (1990) 2466–2470.

    PubMed  Article  CAS  Google Scholar 

  22. Burgess, G.M., Claret, M. and Jenkinson, D.H. Effects of catecholamines, ATP and ionophoree A23187 on potassium and calcium movements in isolated hepatocytes. Nature 279 (1979) 544–546.

    PubMed  Article  CAS  Google Scholar 

  23. Yamamoto, N., Smith, M.W., Maki, A., Berezesky, I.K. and Trump, B.F. Role of cytosolic Ca2+ and protein kinases in the induction of the hsp70 gene. Kidney Int. 45 (1994) 1093–1104.

    PubMed  CAS  Google Scholar 

Download references

Author information

Authors and Affiliations


Corresponding author

Correspondence to Inge Bauer.

Rights and permissions

Reprints and Permissions

About this article

Cite this article

Silomon, M., Bauer, I., Bauer, M. et al. Induction of heme oxygenase-1 and heat shock protein 70 in rat hepatocytes: The role of calcium signaling. Cell Mol Biol Lett 12, 25–38 (2007).

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI:

Key words

  • Stress response genes
  • Hepatocytes
  • Calcium
  • Gene expression