Different Susceptibilities of Osteoclasts and Osteoblasts to Glucocorticoid‑induced Oxidative Stress and Mitochondrial Alterations

10.4103/CJP.CJP_7_19

Yu-Hsu Chen；Shao-Yu Peng；Ming-Te Cheng；Yu-Pao Hsu；Zong-Xi Huang；Winston Teng-Kuei Cheng；Shinn-Chih Wu

Apoptosis ； glucocorticoids ； osteoblasts ； osteoclasts ； oxidative stress

The Chinese Journal of Physiology

62卷2期（2019 / 04 / 30）

70 - 79

英文

Glucocorticoid-induced bone loss is the most common form of secondary osteoporosis. This toxic effect has not been efficiently managed, possibly due to the incomplete understanding of the extraordinarily diverse cellular responses induced by glucocorticoid treatment. Previous literatures revealed that high dose of exogenous glucocorticoid triggers apoptosis in osteocytes and osteoblasts. This cell death is associated with glucocorticoid-induced oxidative stress. In this study, we aimed to investigate the mechanisms of glucocorticoid-induced apoptosis in osteoblasts and examine the responses of osteoclasts to the synthetic glucocorticoid, dexamethasone. We demonstrated the biphasic effects of exogenous glucocorticoid on osteoblastic mitochondrial functions and elevated intracellular oxidative stress in a dose-.and time-dependent manner. On comparison, similar treatment did not induce mitochondrial dysfunctions and oxidative stress in osteoclasts. The production of reactive oxygen/nitrogen species was decreased in osteoclasts. The differences are not due to varying efficiency of cellular antioxidant system. The opposite effects on nitrogen oxide synthase might provide an explanation, as the expression levels of nos2 gene are suppressed in the osteoclast but elevated in the osteoblast. We further revealed that glucocorticoids have a substantial impact on the osteoblastic mitochondria. Basal respiration rate and ATP production were increased upon 24 h incubation of glucocorticoids. The increase in proton leak and nonmitochondrial respiration suggests a potential source of glucocorticoid-induced oxidative stress. Long-term incubation of glucocorticoids accumulates these detrimental changes and results in cytochrome C release and mitochondrial breakdown, consequently leading to apoptosis in osteoblasts. The mitochondrial alterations might be other sources of glucocorticoid-induced oxidative stress in osteoblasts.

1. Almeida, M,Han, L,Ambrogini, E,Weinstein, RS,Manolagas, SC(2011).Glucocorticoids and tumor necrosis factor α increase oxidative stress and suppress Wnt protein signaling in osteoblasts.J Biol Chem,286,44326-44335.
2. Behl, C,Lezoualc’h, F,Trapp, T,Widmann, M,Skutella, T,Holsboer, F(1997).Glucocorticoids enhance oxidative stress-induced cell death in hippocampal neurons in vitro.Endocrinology,138,101-106.
3. Bera, S,Greiner, S,Choudhury, A,Dispenzieri, A,Spitz, DR,Russell, SJ(2010).Dexamethasone-induced oxidative stress enhances myeloma cell radiosensitization while sparing normal bone marrow hematopoiesis.Neoplasia,12,980-992.
4. Buehring, B,Viswanathan, R,Binkley, N,Busse, W(2013).Glucocorticoid-induced osteoporosis: An update on effects and management.J Allergy Clin Immunol,132,1019-1030.
5. Cosman, F,de Beur, SJ,LeBoff, MS,Lewiecki, EM,Tanner, B,Randall, S(2014).Clinician's guide to prevention and treatment of osteoporosis.Osteoporos Int,25,2359-2381.
6. Datta, HK,Ng, WF,Walker, JA,Tuck, SP,Varanasi, SS(2008).The cell biology of bone metabolism.J Clin Pathol,61,577-587.
7. Ding, LZ,Teng, X,Zhang, ZB,Zheng, CJ,Chen, SH(2018).Mangiferin inhibits apoptosis and oxidative stress via BMP2/Smad-1 signaling in dexamethasone-induced MC3T3-E1 cells.Int J Mol Med,41,2517-2526.
8. Du, J,Wang, Y,Hunter, R,Wei, Y,Blumenthal, R,Falke, C(2009).Dynamic regulation of mitochondrial function by glucocorticoids.Proc Natl Acad Sci U S A,106,3543-3548.
9. Eriksen, EF(2010).Cellular mechanisms of bone remodeling.Rev Endocr Metab Disord,11,219-227.
10. Feng, YL,Tang, XL(2014).Effect of glucocorticoid-induced oxidative stress on the expression of cbfa1.Chem Biol Interact,207,26-31.
11. Fraser, LA,Adachi, JD(2009).Glucocorticoid-induced osteoporosis: Treatment update and review.Ther Adv Musculoskelet Dis,1,71-85.
12. Gruver‑Yates, AL,Cidlowski, JA(2013).Tissue‑specific actions of glucocorticoids on apoptosis: A double-edged sword.Cells,2,202-223.
13. Guntur, AR,Gerencser, AA,Le, PT,DeMambro, VE,Bornstein, SA,Mookerjee, SA(2018).Osteoblast-like MC3T3-E1 cells prefer glycolysis for ATP production but adipocyte-like 3T3-L1 cells prefer oxidative phosphorylation.J Bone Miner Res,33,1052-1065.
14. Hachemi, Y,Rapp, AE,Picke, AK,Weidinger, G,Ignatius, A,Tuckermann, J(2018).Molecular mechanisms of glucocorticoids on skeleton and bone regeneration after fracture.J Mol Endocrinol,61,75-90.
15. Hamouda, S,Yasear, A(2009).Effect of dexamethasone on osteoclast formation in the alveolar bone of rabbits.Iraqi J Vet Sci,23,13-16.
16. Han, D,Gao, J,Gu, X,Hengstler, JG,Zhang, L,Shahid, M(2018).P21Waf1/Cip1 depletion promotes dexamethasone-induced apoptosis in osteoblastic MC3T3-E1 cells by inhibiting the Nrf2/HO-1 pathway.Arch Toxicol,92,679-692.
17. Hirayama, T,Sabokbar, A,Athanasou, NA(2002).Effect of corticosteroids on human osteoclast formation and activity.J Endocrinol,175,155-163.
18. Hou, GQ,Guo, C,Song, GH,Fang, N,Fan, WJ,Chen, XD(2013).Lipopolysaccharide (LPS) promotes osteoclast differentiation and activation by enhancing the MAPK pathway and COX-2 expression in RAW264.7 cells.Int J Mol Med,32,503-510.
19. Hsu, E,Nanes, M(2017).Advances in treatment of glucocorticoid-induced osteoporosis.Curr Opin Endocrinol Diabetes Obes,24,411-417.
20. Iuchi, T,Akaike, M,Mitsui, T,Ohshima, Y,Shintani, Y,Azuma, H(2003).Glucocorticoid excess induces superoxide production in vascular endothelial cells and elicits vascular endothelial dysfunction.Circ Res,92,81-87.
21. Jeon, MJ,Kim, JA,Kwon, SH,Kim, SW,Park, KS,Park, SW(2003).Activation of peroxisome proliferator-activated receptor-gamma inhibits the Runx2-mediated transcription of osteocalcin in osteoblasts.J Biol Chem,278,23270-23277.
22. Jia, D,O’Brien, CA,Stewart, SA,Manolagas, SC,Weinstein, RS(2006).Glucocorticoids act directly on osteoclasts to increase their life span and reduce bone density.Endocrinology,147,5592-5599.
23. Johnson, TE,Vogel, R,Rutledge, SJ,Rodan, G,Schmidt, A(1999).Thiazolidinedione effects on glucocorticoid receptor-mediated gene transcription and differentiation in osteoblastic cells.Endocrinology,140,3245-3254.
24. Kearns, AE,Khosla, S,Kostenuik, PJ(2008).Receptor activator of nuclear factor kappaB ligand and osteoprotegerin regulation of bone remodeling in health and disease.Endocr Rev,29,155-192.
25. Khanduja, KL,Kaushik, G,Khanduja, S,Pathak, CM,Laldinpuii, J,Behera, D(2011).Corticosteroids affect nitric oxide generation, total free radicals production, and nitric oxide synthase activity in monocytes of asthmatic patients.Mol Cell Biochem,346,31-37.
26. Maio, R,Perticone, M,Sciacqua, A,Tassone, EJ,Naccarato, P,Bagnato, C(2012).Oxidative stress impairs endothelial function in nondipper hypertensive patients.Cardiovasc Ther,30,85-92.
27. Manolagas, SC(2000).Corticosteroids and fractures: A close encounter of the third cell kind.J Bone Miner Res,15,1001-1005.
28. O’Brien, CA,Jia, D,Plotkin, LI,Bellido, T,Powers, CC,Stewart, SA(2004).Glucocorticoids act directly on osteoblasts and osteocytes to induce their apoptosis and reduce bone formation and strength.Endocrinology,145,1835-1841.
29. Ott, M,Gogvadze, V,Orrenius, S,Zhivotovsky, B(2007).Mitochondria, oxidative stress and cell death.Apoptosis,12,913-922.
30. Owen, R,Reilly, GC(2018).In vitro models of bone remodelling and associated disorders.Front Bioeng Biotechnol,6,134.
31. Plotkin, LI,Weinstein, RS,Parfitt, AM,Roberson, PK,Manolagas, SC,Bellido, T(1999).Prevention of osteocyte and osteoblast apoptosis by bisphosphonates and calcitonin.J Clin Invest,104,1363-1374.
32. Qiao, S,Okret, S,Jondal, M(2009).Thymocyte-synthesized glucocorticoids play a role in thymocyte homeostasis and are down-regulated by adrenocorticotropic hormone.Endocrinology,150,4163-4169.
33. Raggatt, LJ,Partridge, NC(2010).Cellular and molecular mechanisms of bone remodeling.J Biol Chem,285,25103-25108.
34. Rauch, A,Seitz, S,Baschant, U,Schilling, AF,Illing, A,Stride, B(2010).Glucocorticoids suppress bone formation by attenuating osteoblast differentiation via the monomeric glucocorticoid receptor.Cell Metab,11,517-531.
35. Sarinho, ES,Melo, VM(2017).Glucocorticoid-induced bone disease: Mechanisms and importance in pediatric practice.Rev Paul Pediatr,35,207-215.
36. 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,73,60-68.
37. Sato, H,Takahashi, T,Sumitani, K,Takatsu, H,Urano, S(2010).Glucocorticoid generates ROS to induce oxidative injury in the hippocampus, leading to impairment of cognitive function of rats.J Clin Biochem Nutr,47,224-232.
38. Seibel, MJ,Cooper, MS,Zhou, H(2013).Glucocorticoid-induced osteoporosis: Mechanisms, management, and future perspectives.Lancet Diabetes Endocrinol,1,59-70.
39. Shi, XM,Chutkan, N,Hamrick, MW,Isales, CM(2012).Mechanism of Glucocorticoid-Induced Osteoporosis: An Update. Glucocorticoids-New Recognition of Our Familiar Friend.Intech.
40. Siddiqui, JA,Partridge, NC(2016).Physiological bone remodeling: Systemic regulation and growth factor involvement.Physiology,31,233-245.
41. Sivagurunathan, S,Muir, MM,Brennan, TC,Seale, JP,Mason, RS(2005).Influence of glucocorticoids on human osteoclast generation and activity.J Bone Miner Res,20,390-398.
42. Takuma, A,Kaneda, T,Sato, T,Ninomiya, S,Kumegawa, M,Hakeda, Y(2003).Dexamethasone enhances osteoclast formation synergistically with transforming growth factor-beta by stimulating the priming of osteoclast progenitors for differentiation into osteoclasts.J Biol Chem,278,44667-44674.
43. Weijtens, O,Feron, EJ,Schoemaker, RC,Cohen, AF,Lentjes, EG,Romijn, FP(1999).High concentration of dexamethasone in aqueous and vitreous after subconjunctival injection.Am J Ophthalmol,128,192-197.
44. Weinstein, RS,Jilka, RL,Parfitt, AM,Manolagas, SC(1998).Inhibition of osteoblastogenesis and promotion of apoptosis of osteoblasts and osteocytes by glucocorticoids. Potential mechanisms of their deleterious effects on bone.J Clin Invest,102,274-282.
45. Yamada, M,Tsukimura, N,Ikeda, T,Sugita, Y,Att, W,Kojima, N(2013).N-acetyl cysteine as an osteogenesis-enhancing molecule for bone regeneration.Biomaterials,34,6147-6156.
46. You, JM,Yun, SJ,Nam, KN,Kang, C,Won, R,Lee, EH(2009).Mechanism of glucocorticoid-induced oxidative stress in rat hippocampal slice cultures.Can J Physiol Pharmacol,87,440-447.
47. Zhang, W,Yang, N,Shi, XM(2008).Regulation of mesenchymal stem cell osteogenic differentiation by glucocorticoid-induced leucine zipper (GILZ).J Biol Chem,283,4723-4729.
48. Zhou, H,Cooper, MS,Seibel, MJ.(2013).Endogenous glucocorticoids and bone.Bone Res,1,107-119.
49. Zhou, W,Quan, JH,Lee, YH,Shin, DW,Cha, GH(2013).Toxoplasma gondii proliferation require down-regulation of host Nox4 expression via activation of PI3 kinase/Akt signaling pathway.PLoS One,8,e66306.