Low-dose dexamethasone affects osteoblast viability by inducing autophagy via intracellular ROS

Zhang,Shaokun; Liu,Yongyi; Liang,Qingwei

doi:10.3892/mmr.2018.8461

Low-dose dexamethasone affects osteoblast viability by inducing autophagy via intracellular ROS

Authors:
- Shaokun Zhang
- Yongyi Liu
- Qingwei Liang
View Affiliations

Affiliations: Department of Orthopedics, The First Hospital of China Medical University, Shenyang, Liaoning 110000, P.R. China, Department of Sports Medicine, The First Hospital of China Medical University, Shenyang, Liaoning 110000, P.R. China
Published online on: January 18, 2018 https://doi.org/10.3892/mmr.2018.8461
Pages: 4307-4316
Copyright: © Zhang et al. This is an open access article distributed under the terms of Creative Commons Attribution License.

Metrics: Total Views: 0 (Spandidos Publications: | PMC Statistics: )

Metrics: Total PDF Downloads: 0 (Spandidos Publications: | PMC Statistics: )

Cited By (CrossRef): 0 citations Loading Articles...

Abstract

Glucocorticoids (GCs) are closely associated with the progression of GC‑induced osteoporosis (GIOP) by inhibiting osteoblast viability. However, endogenous GCs are important for bone development. In addition, previous studies have demonstrated that GCs could induce autophagy, a cytoprotective process that is protective against various stressors. In the present study, the aim is to explore whether osteoblasts exhibited dose‑dependent viability in the presence of GCs due to autophagy. hFOB 1.19 osteoblasts were treated with various doses of dexamethasone (DEX; 10‑8‑10‑4 M) for 0, 24, 48 and 72 h. The results revealed a biphasic effect of DEX on the viability of hFOB 1.19 cells; a high dose of DEX (≥10‑6 M) accelerated cell apoptosis, while a low dose of DEX (10‑8 M) increased cell viability. Furthermore, significantly increased autophagy was observed in the low dose DEX treatment group, as indicated by the expression of the autophagy‑associated proteins beclin 1 and microtubule‑associated protein light chain 3, and the detection of autophagosomes. Another finding was that DEX upregulated intracellular reactive oxygen species (ROS), which was decreased by the autophagy agonist rapamycin. The increase in autophagy and cell viability associated with low‑dose DEX (10‑8 M) was suppressed by the ROS scavenger catalase and the autophagy inhibitor 3‑methyladenine. In conclusion, the results revealed that GCs affected osteoblast viability in a dose‑dependent manner. A low dose of GCs increased osteoblast viability by inducing autophagy via intracellular ROS. The results indicate that autophagy may be a novel mechanism by which osteoblasts survive GC exposure and provide a potential therapeutic target for treating GIOP.

View Figures

View References

1	Kenanidis E, Potoupnis ME, Kakoulidis P, Leonidou A, Sakellariou GT, Sayegh FE and Tsiridis E: Management of glucocorticoid-induced osteoporosis: Clinical data in relation to disease demographics, bone mineral density and fracture risk. Expert Opin Drug Saf. 14:1035–1053. 2015. View Article : Google Scholar : PubMed/NCBI
2	Frenkel B, White W and Tuckermann J: Glucocorticoid-induced osteoporosis. Adv Exp Med Biol. 872:179–215. 2015. View Article : Google Scholar : PubMed/NCBI
3	Dalle Carbonare L, Arlot ME, Chavassieux PM, Roux JP, Portero NR and Meunier PJ: Comparison of trabecular bone microarchitecture and remodeling in glucocorticoid-induced and postmenopausal osteoporosis. J Bone Miner Res. 16:97–103. 2001. View Article : Google Scholar : PubMed/NCBI
4	Van Staa TP, Laan RF, Barton IP, Cohen S, Reid DM and Cooper C: Bone density threshold and other predictors of vertebral fracture in patients receiving oral glucocorticoid therapy. Arthritis Rheum. 48:3224–3229. 2003. View Article : Google Scholar : PubMed/NCBI
5	Capulli M, Paone R and Rucci N: Osteoblast and osteocyte: Games without frontiers. Arch Biochem Biophys. 561:3–12. 2014. View Article : Google Scholar : PubMed/NCBI
6	Moriishi T and Komori T: Glucocorticoid and bone. The inhibition of osteoblast differentiation and induction of osteocyte apoptosis through the regulation of Bcl-2 by glucocorticoids. Clin Calcium. 24:1329–1336. 2014.(In Japanese).
7	Komori T: Glucocorticoid signaling and bone biology. Horm Metab Res. 48:755–763. 2016. View Article : Google Scholar : PubMed/NCBI
8	Jia J, Yao W, Guan M, Dai W, Shahnazari M, Kar R, Bonewald L, Jiang JX and Lane NE: Glucocorticoid dose determines osteocyte cell fate. FASEB J. 25:3366–3376. 2011. View Article : Google Scholar : PubMed/NCBI
9	Shi J, Wang L, Zhang H, Jie Q, Li X, Shi Q, Huang Q, Gao B, Han Y, Guo K, et al: Glucocorticoids: Dose-related effects on osteoclast formation and function via reactive oxygen species and autophagy. Bone. 79:222–232. 2015. View Article : Google Scholar : PubMed/NCBI
10	Martinez-Lopez N, Athonvarangkul D and Singh R: Autophagy and aging. Adv Exp Med Biol. 847:73–87. 2015. View Article : Google Scholar : PubMed/NCBI
11	Wang JY, Yao WX, Wang Y, Fan YL and Wu JB: Network analysis reveals crosstalk between autophagy genes and disease genes. Sci Rep. 7:443912017. View Article : Google Scholar : PubMed/NCBI
12	Lee JY, Koga H, Kawaguchi Y, Tang W, Wong E, Gao YS, Pandey UB, Kaushik S, Tresse E, Lu J, et al: HDAC6 controls autophagosome maturation essential for ubiquitin-selective quality-control autophagy. EMBO J. 29:969–980. 2010. View Article : Google Scholar : PubMed/NCBI
13	Degenhardt K, Mathew R, Beaudoin B, Bray K, Anderson D, Chen G, Mukherjee C, Shi Y, Gélinas C, Fan Y, et al: Autophagy promotes tumor cell survival and restricts necrosis, inflammation, and tumorigenesis. Cancer Cell. 10:51–64. 2006. View Article : Google Scholar : PubMed/NCBI
14	Sid B, Verrax J and Calderon PB: Role of AMPK activation in oxidative cell damage: Implications for alcohol-induced liver disease. Biochem Pharmacol. 86:200–209. 2013. View Article : Google Scholar : PubMed/NCBI
15	Kouroku Y, Fujita E, Tanida I, Ueno T, Isoai A, Kumagai H, Ogawa S, Kaufman RJ, Kominami E and Momoi T: ER stress (PERK/eIF2alpha phosphorylation) mediates the polyglutamine-induced LC3 conversion, an essential step for autophagy formation. Cell Death Differ. 14:230–239. 2007. View Article : Google Scholar : PubMed/NCBI
16	Gu X, Han D, Chen W, Zhang L, Lin Q, Gao J, Fanning S and Han B: SIRT1-mediated FoxOs pathways protect against apoptosis by promoting autophagy in osteoblast-like MC3T3-E1 cells exposed to sodium fluoride. Oncotarget. 7:65218–65230. 2016. View Article : Google Scholar : PubMed/NCBI
17	Yang L, Meng H and Yang M: Autophagy protects osteoblasts from advanced glycation end products-induced apoptosis through intracellular reactive oxygen species. J Mol Endocrinol. 56:291–300. 2016. View Article : Google Scholar : PubMed/NCBI
18	Klionsky DJ, Abdelmohsen K, Abe A, Abedin MJ, Abeliovich H, Acevedo Arozena A, Adachi H, Adams CM, Adams PD, Adeli K, et al: Guidelines for the use and interpretation of assays for monitoring autophagy (3rd edition). Autophagy. 12:1–222. 2016. View Article : Google Scholar : PubMed/NCBI
19	Tanida I, Ueno T and Kominami E: LC3 and autophagy. Methods Mol Biol. 445:77–88. 2008. View Article : Google Scholar : PubMed/NCBI
20	Fu LL, Cheng Y and Liu B: Beclin-1: Autophagic regulator and therapeutic target in cancer. Int J Biochem Cell Biol. 45:921–924. 2013. View Article : Google Scholar : PubMed/NCBI
21	Grishko V, Xu M, Ho R, Mates A, Watson S, Kim JT, Wilson GL and Pearsall AW IV: Effects of hyaluronic acid on mitochondrial function and mitochondria-driven apoptosis following oxidative stress in human chondrocytes. J Biol Chem. 284:9132–9139. 2009. View Article : Google Scholar : PubMed/NCBI
22	Ong SL, Zhang Y and Whitworth JA: Reactive oxygen species and glucocorticoid-induced hypertension. Clin Exp Pharmacol Physiol. 35:477–482. 2008. View Article : Google Scholar : PubMed/NCBI
23	Henrotin Y, Kurz B and Aigner T: Oxygen and reactive oxygen species in cartilage degradation: Friends or foes? Osteoarthritis Cartilage. 13:643–654. 2005. View Article : Google Scholar : PubMed/NCBI
24	Ghavami S, Shojaei S, Yeganeh B, Ande SR, Jangamreddy JR, Mehrpour M, Christoffersson J, Chaabane W, Moghadam AR, Kashani HH, et al: Autophagy and apoptosis dysfunction in neurodegenerative disorders. Prog Neurobiol. 112:24–49. 2014. View Article : Google Scholar : PubMed/NCBI
25	Almeida M, Han L, Martin-Millan M, Plotkin LI, Stewart SA, Roberson PK, Kousteni S, O'Brien CA, Bellido T, Parfitt AM, et al: Skeletal involution by age-associated oxidative stress and its acceleration by loss of sex steroids. J Biol Chem. 282:27285–27297. 2007. View Article : Google Scholar : PubMed/NCBI
26	Wauquier F, Leotoing L, Coxam V, Guicheux J and Wittrant Y: Oxidative stress in bone remodelling and disease. Trends Mol Med. 15:468–477. 2009. View Article : Google Scholar : PubMed/NCBI
27	Harris SA, Enger RJ, Riggs BL and Spelsberg TC: Development and characterization of a conditionally immortalized human fetal osteoblastic cell line. J Bone Miner Res. 10:178–186. 1995. View Article : Google Scholar : PubMed/NCBI
28	Brand MD, Affourtit C, Esteves TC, Green K, Lambert AJ, Miwa S, Pakay JL and Parker N: Mitochondrial superoxide: Production, biological effects, and activation of uncoupling proteins. Free Radic Biol Med. 37:755–767. 2004. View Article : Google Scholar : PubMed/NCBI
29	Wohlgemuth SE, Calvani R and Marzetti E: The interplay between autophagy and mitochondrial dysfunction in oxidative stress-induced cardiac aging and pathology. J Mol Cell Cardiol. 71:62–70. 2014. View Article : Google Scholar : PubMed/NCBI
30	Mizushima N, Levine B, Cuervo AM and Klionsky DJ: Autophagy fights disease through cellular self-digestion. Nature. 451:1069–1075. 2008. View Article : Google Scholar : PubMed/NCBI
31	Kalak R, Zhou H, Street J, Day RE, Modzelewski JR, Spies CM, Liu PY, Li G, Dunstan CR and Seibel MJ: Endogenous glucocorticoid signalling in osteoblasts is necessary to maintain normal bone structure in mice. Bone. 45:61–67. 2009. View Article : Google Scholar : PubMed/NCBI
32	Zhou H, Mak W, Kalak R, Street J, Fong-Yee C, Zheng Y, Dunstan CR and Seibel MJ: Glucocorticoid-dependent Wnt signaling by mature osteoblasts is a key regulator of cranial skeletal development in mice. Development. 136:427–436. 2009. View Article : Google Scholar : PubMed/NCBI
33	Ishida Y, Tertinegg I and Heersche JN: Progesterone and dexamethasone stimulate proliferation and differentiation of osteoprogenitors and progenitors for adipocytes and macrophages in cell populations derived from adult rat vertebrae. J Bone Miner Res. 11:921–930. 1996. View Article : Google Scholar : PubMed/NCBI
34	Atmani H, Chappard D and Basle MF: Proliferation and differentiation of osteoblasts and adipocytes in rat bone marrow stromal cell cultures: Effects of dexamethasone and calcitriol. J Cell Biochem. 89:364–372. 2003. View Article : Google Scholar : PubMed/NCBI
35	Wang F, Jia J and Rodrigues B: Autophagy, metabolic disease, and pathogenesis of heart dysfunction. Can J Cardiol. 33:850–859. 2017. View Article : Google Scholar : PubMed/NCBI
36	Booth LA, Tavallai S, Hamed HA, Cruickshanks N and Dent P: The role of cell signalling in the crosstalk between autophagy and apoptosis. Cell Signal. 26:549–555. 2014. View Article : Google Scholar : PubMed/NCBI
37	Bhutia SK, Kegelman TP, Das SK, Azab B, Su ZZ, Lee SG, Sarkar D and Fisher PB: Astrocyte elevated gene-1 induces protective autophagy. Proc Natl Acad Sci USA. 107:pp. 22243–22248. 2010; View Article : Google Scholar : PubMed/NCBI
38	Ouyang L, Shi Z, Zhao S, Wang FT, Zhou TT, Liu B and Bao JK: Programmed cell death pathways in cancer: A review of apoptosis, autophagy and programmed necrosis. Cell Prolif. 45:487–498. 2012. View Article : Google Scholar : PubMed/NCBI
39	Bhutia SK, Mukhopadhyay S, Sinha N, Das DN, Panda PK, Patra SK, Maiti TK, Mandal M, Dent P, Wang XY, et al: Autophagy: Cancer's friend or foe? Adv Cancer Res. 118:61–95. 2013. View Article : Google Scholar : PubMed/NCBI
40	Manolagas SC and Parfitt AM: What old means to bone. Trends Endocrinol Metab. 21:369–374. 2010. View Article : Google Scholar : PubMed/NCBI
41	Lv XH, Zhao DH, Cai SZ, Luo SY, You T, Xu BL and Chen K: Autophagy plays a protective role in cell death of osteoblasts exposure to lead chloride. Toxicol Lett. 239:131–140. 2015. View Article : Google Scholar : PubMed/NCBI
42	Wang L, Fan J, Lin YS, Guo YS, Gao B, Shi QY, Wei BY, Chen L, Yang L, Liu J and Luo ZJ: Glucocorticoids induce autophagy in rat bone marrow mesenchymal stem cells. Mol Med Rep. 11:2711–2716. 2015. View Article : Google Scholar : PubMed/NCBI
43	Zhao Y, Zuo Y, Huo HJ, Xiao YL, Yang XJ and Xin DQ: Glucocorticoid induced autophagy in N1511 chondrocyte cells. Eur Rev Med Pharmacol Sci. 18:3573–3579. 2014.PubMed/NCBI
44	Grander D, Kharaziha P, Laane E, Pokrovskaja K and Panaretakis T: Autophagy as the main means of cytotoxicity by glucocorticoids in hematological malignancies. Autophagy. 5:1198–1200. 2009. View Article : Google Scholar : PubMed/NCBI
45	Almeida M, Han L, Ambrogini E, Weinstein RS and Manolagas SC: Glucocorticoids and tumor necrosis factor alpha increase oxidative stress and suppress Wnt protein signaling in osteoblasts. J Biol Chem. 286:44326–44335. 2011. View Article : Google Scholar : PubMed/NCBI
46	Kiffin R, Bandyopadhyay U and Cuervo AM: Oxidative stress and autophagy. Antioxid Redox Signal. 8:152–162. 2006. View Article : Google Scholar : PubMed/NCBI
47	Zhen YF, Wang GD, Zhu LQ, Tan SP, Zhang FY, Zhou XZ and Wang XD: P53 dependent mitochondrial permeability transition pore opening is required for dexamethasone-induced death of osteoblasts. J Cell Physiol. 229:1475–1483. 2014. View Article : Google Scholar : PubMed/NCBI
48	Manolagas SC: From estrogen-centric to aging and oxidative stress: A revised perspective of the pathogenesis of osteoporosis. Endocr Rev. 31:266–300. 2010. View Article : Google Scholar : PubMed/NCBI
49	Schroder K: NADPH oxidases in bone homeostasis and osteoporosis. Cell Mol Life Sci. 72:25–38. 2015. View Article : Google Scholar : PubMed/NCBI
50	Suzuki M, Bandoski C and Bartlett JD: Fluoride induces oxidative damage and SIRT1/autophagy through ROS-mediated JNK signaling. Free Radic Biol Med. 89:369–378. 2015. View Article : Google Scholar : PubMed/NCBI
51	Scherz-Shouval R and Elazar Z: ROS, mitochondria and the regulation of autophagy. Trends Cell Biol. 17:422–427. 2007. View Article : Google Scholar : PubMed/NCBI