Advertisement

Heme oxygenase-1 prevents glucocorticoid and hypoxia-induced apoptosis and necrosis of osteocyte-like cells

  • Hiroki Yamamoto
  • Masazumi SaitoEmail author
  • Tsuyoshi Goto
  • Keiichiro Ueshima
  • Masashi Ishida
  • Shigeki Hayashi
  • Kazuya Ikoma
  • Osam Mazda
  • Toshikazu Kubo
Original Paper
  • 24 Downloads

Abstract

Glucocorticoids and hypoxia is considered to promote osteocyte apoptosis and necrosis, which are observed in glucocorticoid-associated osteonecrosis and osteoporosis. Heme oxygenase-1 (HO-1) induced by hemin is reported to have cytoprotective effects in ischemic diseases. The objective of this study was to evaluate the effect of HO-1 on osteocyte death caused by glucocorticoids and hypoxia. We confirmed that hemin induced HO-1 expression in MLO-Y4 mouse osteocytes. MLO-Y4 was cultured with dexamethasone (Dex) under hypoxia (DH group). Furthermore, these cells were cultured with hemin (DH-h group) or hemin and zinc protoporphyrin IX (an HO-1 inhibitor) (DH-h-PP group). The rates of apoptosis and necrosis of these groups were analyzed by flow cytometry and compared with cells cultured under normal condition. Both apoptosis and necrosis increased in the DH group. Hemin administration significantly reduced cell death caused by glucocorticoids and hypoxia in the DH-h group, and its effect was attenuated by the HO-1 inhibitor in DH-h-PP group. Capase-3 activity significantly decreased in the DH-h group. This implied that the cell death inhibition effect due to hemin is mediated by HO-1 and caspase-3. HO-1 induction may be useful in the treatment of glucocorticoid-associated osteonecrosis and osteoporosis.

Keywords

Heme oxygenase-1 Hemin Osteonecrosis of the femoral head Osteoporosis Hypoxia 

Notes

Funding

JSPS KAKENHI; Grant Number: 15K20011.

Compliance with Ethical Standards

Conflict of interest

The authors declare that they have no conflicts of interest.

References

  1. 1.
    Mankin HJ (1992) Nontraumatic necrosis of bone (osteonecrosis). N Engl J Med 326:1473–1479CrossRefGoogle Scholar
  2. 2.
    Jones JP Jr (1994) Concepts of etiology and early pathogenesis of osteonecrosis. Instr Course Lect 43:499–512Google Scholar
  3. 3.
    Jones JP Jr (1993) Fat embolism, intravascular coagulation, and osteonecrosis. Clin Orthop Relat Res 292:294–308Google Scholar
  4. 4.
    Fisher DE (1978) The role of fat embolism in the etiology of corticosteroid-induced avascular necrosis: clinical and experimental results. Clin Orthop Relat Res 130:68–80Google Scholar
  5. 5.
    Motomura G, Yamamoto T, Miyanishi K, Jingushi S, Iwamoto Y (2004) Combined effects of an anticoagulant and a lipid-lowering agent on the prevention of steroid-induced osteonecrosis in rabbits. Arthritis Rheum 50:3387–3391CrossRefGoogle Scholar
  6. 6.
    Kabata T, Matsumoto T, Yagishita S, Wakayama T, Iseki S, Tomita K (2008) Vascular endothelial growth factor in rabbits during development of corticosteroid-induced osteonecrosis: a controlled experiment. J Rheumatol 35:2383–2390CrossRefGoogle Scholar
  7. 7.
    Wang GJ, Sweet DE, Reger SI, Thompson RC (1977) Fat-cell changes as a mechanism of avascular necrosis of the femoral head in cortisone-treated rabbits. J Bone Joint Surg Am 59:729–735CrossRefGoogle Scholar
  8. 8.
    Wang TY, Avlonitis EG, Relkin R (1988) Systemic necrotizing vasculitis causing bone necrosis. Am J Med 84:1085–1086CrossRefGoogle Scholar
  9. 9.
    Saito S, Inoue A, Ono K (1987) Intramedullary haemorrhage as a possible cause of avascular necrosis of the femoral head. The histology of 16 femoral heads at the silent stage. J Bone Joint Surg Br 69:346–351CrossRefGoogle Scholar
  10. 10.
    Kerachian MA, Cournoyer D, Harvey EJ, Chow TY, Neagoe PE, Sirois MG, Seguin C (2009) Effect of high-dose dexamethasone on endothelial haemostatic gene expression and neutrophil adhesion. J Steroid Biochem Mol Biol 116:127–133CrossRefGoogle Scholar
  11. 11.
    Ichiseki T, Matsumoto T, Nishino M, Kaneuji A, Katsuda S (2004) Oxidative stress and vascular permeability in steroid-induced osteonecrosis model. J Orthop Sci 9:509–515CrossRefGoogle Scholar
  12. 12.
    Luo Y, Liu X, Zheng Q, Wan X, Ouyang S, Yin Y, Sui X, Liu J, Yang X (2012) Hydrogen sulfide prevents hypoxia-induced apoptosis via inhibition of an H2O2-activated calcium signaling pathway in mouse hippocampal neurons. Biochem Biophys Res Commun 425:473–477CrossRefGoogle Scholar
  13. 13.
    Kerachian MA, Harvey EJ, Cournoyer D, Chow TYK, Séguin C (2006) Avascular necrosis of the femoral head: Vascular hypotheses. Endothelium 13:237–244CrossRefGoogle Scholar
  14. 14.
    Zhao ZQ, Bai R, Liu WL, Feng W, Zhao AQ, Wang Y, Wang WX, Sun L, Wu LS, Cui SX (2016) Roles of oxidative DNA damage of bone marrow hematopoietic cells in steroid-induced avascular necrosis of femoral head. Genet Mol Res 24(1):15Google Scholar
  15. 15.
    Zhu CT, Li T, Hu YH, Zou M, Guo Q, QU XW (2017) Exosomes secreted by mice adipose-derived stem cells after low-level laser irradiation treatment reduce apoptosis of osteocyte induced by hypoxia. Eur Rev Med Pharmacol Sci 21:5562–5570Google Scholar
  16. 16.
    Han D, Gao J, Gu X, Hengstler JG, Zhang L, Shahid M, Ali T, Han B (2017) P21Waf1/Cip1 depletion promotes dexamethasoneinduced apoptosis in osteoblastic MC3T3E1 cells by inhibiting the Nrf2/HO1 pathway. Arch Toxicol 92:679–692CrossRefGoogle Scholar
  17. 17.
    Wu J, Yao L, Wang B, Liu Z, Ma K (2016) Tao-Hong-Si-Wu Decoction ameliorates steroid-induced avascular necrosis of the femoral head by regulating the HIF-1α pathway and cell apoptosis. Biosci Trends 10:410–417CrossRefGoogle Scholar
  18. 18.
    Ueda S, Ichiseki T, Yoshitomi Y, Yonekura H, Ueda Y, Kaneuji A, Matsumoto T (2015) Osteocytic cell necrosis is caused by a combination of glucocorticoid-induced Dickkopf-1 and hypoxia. Med Mol Morphol 48:69–75CrossRefGoogle Scholar
  19. 19.
    Balla G, Jacob HS, Balla J, Rosenberg M, Nath K, Apple F, Eaton JW, Vercellotti GM (1992) Ferritin: a cytoprotective antioxidant strategem of endothelium. J Biol Chem 267:18148–18153Google Scholar
  20. 20.
    Idelman G, Smith DL, Zucker SD (2015) Bilirubin inhibits the up-regulation of inducible nitric oxide synthase by scavenging reactive oxygen species generated by the toll-like receptor 4-dependent activation of NADPH oxidase. Redox Biol 5:398–408CrossRefGoogle Scholar
  21. 21.
    Ryter SW, Choi AM (2016) Targeting heme oxygenase-1 and carbon monoxide for therapeutic modulation of inflammation. Transl Res 167:7–34CrossRefGoogle Scholar
  22. 22.
    Hangaishi M, Ishizaka N, Aizawa T, Kurihara Y, Taguchi J, Nagai R, Kimura S, Ohno M (2000) Induction of heme oxygenase-1 can act protectively against cardiac ischemia/reperfusion in vivo. Biochem Biophys Res Commun 279:582–588CrossRefGoogle Scholar
  23. 23.
    Zhu J, Zhang L, Wu X, Xiong Z, Qiu Y, Hua T, Tang G (2017) Reduction of longitudinal, vertebral blood perfusion and its likely causes: a quantitative dynamic contrast-enhanced MR imaging study of a rat osteoporosis model. Radiology 282:369–380CrossRefGoogle Scholar
  24. 24.
    Kato Y, Windle JJ, Koop BA, Mundy GR, Bonewald LF (1997) Establishment of an osteocyte-like cell line, MLO-Y4. J Bone Miner Res 12:2014–2023CrossRefGoogle Scholar
  25. 25.
    Jähn K, Lara-Castillo N, Brotto L, Mo CL, Johnson ML, Brotto M, Bonewald LF (2012) Skeletal muscle secreted factors prevent glucocorticoid-induced osteocyte apoptosis through activation of b-catenin. Eur Cell Mater 24:197–209CrossRefGoogle Scholar
  26. 26.
    Xia X, Kar R, Gluhak-Heinrich J, Yao W, Lane NE, Bonewald LF, Biswas SK, Lo WK, Jiang JX (2010) Glucocorticoid-induced autophagy in osteocytes. J Bone Miner Res 25:2479–2488CrossRefGoogle Scholar
  27. 27.
    Tsuji M, Ikeda H, Ishizu A, Miyatake Y, Hayase H, Yoshiki T (2006) Altered expression of apoptosis-related genes in osteocytes exposed to high-dose steroid hormones and hypoxic stress. Pathobiology 73:304–309CrossRefGoogle Scholar
  28. 28.
    Ingraham CA, Park GC, Makarenkova HP, Crossin KL (2011) Matrix metalloproteinase (MMP)-9 induced by Wnt signaling increases the proliferation and migration of embryonic neural stem cells at low O2 levels. J Biol Chem 286:17649–17657CrossRefGoogle Scholar
  29. 29.
    Xiao Q, Piao R, Wang H, Li C, Song L (2018) Orientin-mediated Nrf2/HO-1 signal alleviates H2O2-induced oxidative damage via induction of JNK and PI3K/AKT activation. Int J Biol Macromol 118:747–755CrossRefGoogle Scholar
  30. 30.
    Ndisang JF, Jadhav A, Mishra M (2014) The heme oxygenase system suppresses perirenal visceral adiposity, abates renal inflammation and ameliorates diabetic nephropathy in Zucker diabetic fatty rats. PLoS One 9:e87936CrossRefGoogle Scholar
  31. 31.
    Rossi M, Thierry A, Delbauve S, Preyat N, Soares MP, Roumeguère T, Leo O, Flamand V, Le Moine A, Hougardy JM (2017) Specific expression of heme oxygenase-1 by myeloid cells modulates renal ischemia-reperfusion injury. Sci Rep 7:197CrossRefGoogle Scholar
  32. 32.
    Kabata T, Kubo T, Matsumoto T, Nishino M, Tomita K, Katsuda S, Horii T, Uto N, Kitajima I (2000) Apoptotic cell death in steroid induced osteonecrosis: an experimental study in rabbits. J Rheumatol 27:2166–2171Google Scholar
  33. 33.
    O’Brien CA, Jia D, Plotkin LI, Bellido T, Powers CC, Stewart SA, Manolagas SC, Weinstein RS (2004) Glucocorticoids act directly on osteoblasts and osteocytes to induce their apoptosis and reduce bone formation and strength. Endocrinology 145:1835–1841CrossRefGoogle Scholar
  34. 34.
    Kogianni G, Mann V, Ebetino F, Nuttall M, Nijweide P, Simpson H, Noble B (2004) Fas/CD95 is associated with glucocorticoid-induced osteocyte apoptosis. Life Sci 75:2879–2895CrossRefGoogle Scholar
  35. 35.
    Frenkel B, White W, Tuckermann J (2016) Glucocorticoid-induced osteoporosis. Adv Exp Med Biol 872:179–215CrossRefGoogle Scholar
  36. 36.
    Akaike M, Matsumoto T (2007) Glucocorticoid-induced reduction in NO bioavailability and vascular endothelial dysfunction. Clin calcium 17:864–870Google Scholar
  37. 37.
    Okada Y, Tanikawa T, Iida T, Tanaka Y (2007) Vascular injury by glucocorticoid; involvement of apoptosis of endothelial cells. Clin calcium 17:872–877Google Scholar
  38. 38.
    Lin H, Gao X, Chen G, Sun J, Chu J, Jing K, Li P, Zeng R, Wei B (2015) Indole-3-carbinol as inhibitors of glucocorticoid-induced apoptosis in osteoblastic cells through blocking ROS-mediated Nrf2 pathway. Biochem Biophys Res Commun 46:422–427CrossRefGoogle Scholar
  39. 39.
    Sato AY, Tu X, McAndrews KA, Plotkin LI, Bellido T (2015) Prevention of glucocorticoid induced-apoptosis of osteoblasts and osteocytes by protecting against endoplasmic reticulum (ER) stress in vitro and in vivo in female mice. Bone 73:60–68CrossRefGoogle Scholar

Copyright information

© The Japanese Society for Clinical Molecular Morphology 2019

Authors and Affiliations

  • Hiroki Yamamoto
    • 1
  • Masazumi Saito
    • 1
    Email author
  • Tsuyoshi Goto
    • 1
  • Keiichiro Ueshima
    • 1
  • Masashi Ishida
    • 1
  • Shigeki Hayashi
    • 1
  • Kazuya Ikoma
    • 1
  • Osam Mazda
    • 2
  • Toshikazu Kubo
    • 1
  1. 1.Department of Orthopaedics, Graduate School of Medical ScienceKyoto Prefectural University of MedicineKyotoJapan
  2. 2.Department of Immunology, Graduate School of Medical ScienceKyoto Prefectural University of MedicineKyotoJapan

Personalised recommendations