Top

BMC Musculoskeletal Disorders

Published in:

Open Access 01-12-2023 | Erythropoietin | Research

Erythropoietin suppresses osteoblast apoptosis and ameliorates steroid-induced necrosis of the femoral head in rats by inhibition of STAT1-caspase 3 signaling pathway

Authors: Tingwen Cai, Siyuan Chen, Chenghu Wu, Chao Lou, Weidan Wang, Chihao Lin, Hongyi Jiang, Xinxian Xu

Published in: BMC Musculoskeletal Disorders | Issue 1/2023

Abstract

Background

Steroid-induced avascular necrosis of the femoral head (SANFH) is characterized by osteoblast apoptosis, leading to a loss of bone structure and impaired hip joint function. It has been demonstrated that erythropoietin (EPO) performs a number of biological roles.

Objective

We examined the effects of EPO on SANFH and its regulation of the STAT1-caspase 3 signaling pathway.

Method

In vitro, osteoblasts were treated with dexamethasone (Dex) or EPO. We identified the cytotoxicity of EPO by CCK-8, the protein expression of P-STAT1, cleaved-caspase9, cleaved-caspase3, Bcl-2, BAX, and cytochrome c by Western blotting, and evaluated the apoptosis of osteoblasts by flow cytometry. In vivo, we analyzed the protective effect of EPO against SANFH by hematoxylin and eosin (H&E), Immunohistochemical staining, and Micro-computed tomography (CT).

Results

In vitro, EPO had no apparent toxic effect on osteoblasts. In Dex-stimulated cells, EPO therapy lowered the protein expression of BAX, cytochrome c, p-STAT1, cleaved-caspase9, and cleaved-caspase3 while increasing the expression of Bcl-2. EPO can alleviate the apoptosis induced by Dex. In vivo, EPO can lower the percentage of empty bone lacunae in SANFH rats.

Conclusion

The present study shows that EPO conferred beneficial effects in rats with SANFH by inhibiting STAT1-caspase 3 signaling, suggesting that EPO may be developed as a treatment for SANFH.

Available only for authorised users

Venkatesh B, Finfer S, Cohen J, et al. Adjunctive glucocorticoid therapy in patients with septic shock. N Engl J Med. 2018;378(9):797–808. https://doi.org/10.1056/NEJMoa1705835.CrossRefPubMed

Liu LH, Zhang QY, Sun W, Li ZR, Gao FQ. Corticosteroid-induced osteonecrosis of the femoral head: detection, diagnosis, and treatment in earlier stages. Chin Med J (Engl). 2017;130(21):2601–7. https://doi.org/10.4103/0366-6999.217094.CrossRefPubMed

Chang C, Greenspan A, Gershwin ME. The pathogenesis, diagnosis and clinical manifestations of steroid-induced osteonecrosis. J Autoimmun. 2020;110:102460. https://doi.org/10.1016/j.jaut.2020.102460.CrossRefPubMed

Kerachian MA, Séguin C, Harvey EJ. Glucocorticoids in osteonecrosis of the femoral head: a new understanding of the mechanisms of action. J Steroid Biochem Mol Biol. 2009;114(3–5):121–8. https://doi.org/10.1016/j.jsbmb.2009.02.007.CrossRefPubMedPubMedCentral

Komori T. Cell death in Chondrocytes, osteoblasts, and Osteocytes. Int J Mol Sci. 2016;17(12):E2045. https://doi.org/10.3390/ijms17122045.CrossRef

Cooper MS, Seibel MJ, Zhou H. Glucocorticoids, bone and energy metabolism. Bone. 2016;82:64–8. https://doi.org/10.1016/j.bone.2015.05.038.CrossRefPubMed

Brennan-Speranza TC, Henneicke H, Gasparini SJ, et al. Osteoblasts mediate the adverse effects of glucocorticoids on fuel metabolism. J Clin Invest. 2012;122(11):4172–89. https://doi.org/10.1172/JCI63377.CrossRefPubMedPubMedCentral

Verhoeven Y, Tilborghs S, Jacobs J, et al. The potential and controversy of targeting STAT family members in cancer. Semin Cancer Biol. 2020;60:41–56. https://doi.org/10.1016/j.semcancer.2019.10.002.CrossRefPubMed

Dodington DW, Desai HR, Woo M. JAK/STAT - emerging players in metabolism. Trends Endocrinol Metab. 2018;29(1):55–65. https://doi.org/10.1016/j.tem.2017.11.001.CrossRefPubMed

10.

Martí-Rodrigo A, Alegre F, Moragrega ÁB, et al. Rilpivirine attenuates liver fibrosis through selective STAT1-mediated apoptosis in hepatic stellate cells. Gut. 2020;69(5):920–32. https://doi.org/10.1136/gutjnl-2019-318372.CrossRefPubMed

11.

Huang F, Wang Q, Guo F, et al. FoxO1-mediated inhibition of STAT1 alleviates tubulointerstitial fibrosis and tubule apoptosis in diabetic Kidney Disease. EBioMedicine. 2019;48:491–504. https://doi.org/10.1016/j.ebiom.2019.09.002.CrossRefPubMedPubMedCentral

12.

Porter AG, Jänicke RU. Emerging roles of caspase-3 in apoptosis. Cell Death Differ. 1999;6(2):99–104. https://doi.org/10.1038/sj.cdd.4400476.CrossRefPubMed

13.

Feng Z, Zheng W, Tang Q, et al. Fludarabine inhibits STAT1-mediated up-regulation of caspase-3 expression in dexamethasone-induced osteoblasts apoptosis and slows the progression of steroid-induced avascular necrosis of the femoral head in rats. Apoptosis. 2017;22(8):1001–12. https://doi.org/10.1007/s10495-017-1383-1.CrossRefPubMed

14.

Xu X, Wen H, Hu Y, et al. STAT1-caspase 3 pathway in the apoptotic process associated with steroid-induced necrosis of the femoral head. J Mol Histol. 2014;45(4):473–85. https://doi.org/10.1007/s10735-014-9571-6.CrossRefPubMed

15.

Kao R, Xenocostas A, Rui T, et al. Erythropoietin improves skeletal muscle microcirculation and tissue bioenergetics in a mouse sepsis model. Crit Care. 2007;11(3):R58. https://doi.org/10.1186/cc5920.CrossRefPubMedPubMedCentral

16.

Yang B, Hosgood SA, Bagul A, Waller HL, Nicholson ML. Erythropoietin regulates apoptosis, inflammation and tissue remodelling via caspase-3 and IL-1β in isolated hemoperfused kidneys. Eur J Pharmacol. 2011;660(2–3):420–30. https://doi.org/10.1016/j.ejphar.2011.03.044.CrossRefPubMed

17.

Nairz M, Sonnweber T, Schroll A, Theurl I, Weiss G. The pleiotropic effects of erythropoietin in Infection and inflammation. Microbes Infect. 2012;14(3):238–46. https://doi.org/10.1016/j.micinf.2011.10.005.CrossRefPubMedPubMedCentral

18.

Peng B, Kong G, Yang C, Ming Y. Erythropoietin and its derivatives: from tissue protection to immune regulation. Cell Death Dis. 2020;11(2):79. https://doi.org/10.1038/s41419-020-2276-8.CrossRefPubMedPubMedCentral

19.

Kirito K, Nakajima K, Watanabe T, et al. Identification of the human erythropoietin receptor region required for Stat1 and Stat3 activation. Blood. 2002;99(1):102–10. https://doi.org/10.1182/blood.v99.1.102.CrossRefPubMed

20.

Jiang C, Xu Q, Xu K, et al. Effects of erythropoietin on STAT1 and STAT3 levels following cerebral ischemia-reperfusion in rats. Int J Neurosci. 2013;123(10):684–90. https://doi.org/10.3109/00207454.2013.817409.CrossRefPubMed

21.

Chen S, Li J, Peng H, Zhou J, Fang H. Administration of erythropoietin exerts protective effects against glucocorticoid-induced osteonecrosis of the femoral head in rats. Int J Mol Med. 2014;33(4):840–8. https://doi.org/10.3892/ijmm.2014.1644.CrossRefPubMedPubMedCentral

22.

Li D, Xie X, Yang Z, Wang C, Wei Z, Kang P. Enhanced bone defect repairing effects in glucocorticoid-induced osteonecrosis of the femoral head using a porous nano-lithium-hydroxyapatite/gelatin microsphere/erythropoietin composite scaffold. Biomater Sci. 2018;6(3):519–37. https://doi.org/10.1039/c7bm00975e.CrossRefPubMed

23.

Low SC, Bain GI, Findlay DM, Eng K, Perilli E. External and internal bone micro-architecture in normal and Kienböck’s lunates: a whole-bone micro-computed tomography study. J Orthop Res. 2014;32(6):826–33. https://doi.org/10.1002/jor.22611.CrossRefPubMed

24.

Dong Y, Li Y, Huang C, Gao K, Weng X. Systemic application of teriparatide for steroid induced osteonecrosis in a rat model. BMC Musculoskelet Disord. 2015;16:163. https://doi.org/10.1186/s12891-015-0589-z.CrossRefPubMedPubMedCentral

25.

Mont MA, Jones LC, Hungerford DS. Nontraumatic osteonecrosis of the femoral head: ten years later. J Bone Joint Surg Am. 2006;88(5):1117–32. https://doi.org/10.2106/JBJS.E.01041.CrossRefPubMed

26.

Mont MA, Jones LC, Einhorn TA, Hungerford DS, Reddi AH. Osteonecrosis of the femoral head. Potential treatment with growth and differentiation factors. Clin Orthop Relat Res. 1998;(355 Suppl):S314–335.

27.

Yu H, Liu P, Zuo W, et al. Decreased angiogenic and increased apoptotic activities of bone microvascular endothelial cells in patients with glucocorticoid-induced osteonecrosis of the femoral head. BMC Musculoskelet Disord. 2020;21(1):277. https://doi.org/10.1186/s12891-020-03225-1.CrossRefPubMedPubMedCentral

28.

Zalavras C, Shah S, Birnbaum MJ, Frenkel B. Role of apoptosis in glucocorticoid-induced osteoporosis and osteonecrosis. Crit Rev Eukaryot Gene Expr. 2003;13(2–4):221–35.PubMed

29.

Wang XY, Hua BX, Jiang C, et al. Serum biomarkers related to glucocorticoid-Induced osteonecrosis of the femoral head: a prospective nested case-control study. J Orthop Res. 2019;37(11):2348–57. https://doi.org/10.1002/jor.24400.CrossRefPubMed

30.

Nie Z, Chen S, Peng H. Glucocorticoid induces osteonecrosis of the femoral head in rats through GSK3β-mediated osteoblast apoptosis. Biochem Biophys Res Commun. 2019;511(3):693–9. https://doi.org/10.1016/j.bbrc.2019.02.118.CrossRefPubMed

31.

Wang C, Wang X, Xu X. Bone microstructure and regional distribution of osteoblast and osteoclast activity in the osteonecrotic femoral head. PLoS ONE. 2014;9(5):e96361. https://doi.org/10.1371/journal.pone.0096361.CrossRefPubMedPubMedCentral

32.

Tajima K, Takaishi H, Takito J, et al. Inhibition of STAT1 accelerates bone fracture healing. J Orthop Res. 2010;28(7):937–41. https://doi.org/10.1002/jor.21086.CrossRefPubMed

33.

Kostanian IA, Vonarshenko AV, Lipkin VM. [STAT1: a many-sided transcription factor]. Bioorg Khim. 2010;36(1):15–28.PubMed

34.

Savill J, Fadok V. Corpse clearance defines the meaning of cell death. Nature. 2000;407(6805):784–8. https://doi.org/10.1038/35037722.CrossRefPubMed

35.

Sisler JD, Morgan M, Raje V, et al. The Signal Transducer and activator of transcription 1 (STAT1) inhibits mitochondrial Biogenesis in liver and fatty acid oxidation in adipocytes. PLoS ONE. 2015;10(12):e0144444. https://doi.org/10.1371/journal.pone.0144444.CrossRefPubMedPubMedCentral

36.

Dietrich JB. [Apoptosis and anti-apoptosis genes in the Bcl-2 family]. Arch Physiol Biochem. 1997;105(2):125–35. https://doi.org/10.1076/apab.105.2.125.12927.CrossRefPubMed

37.

Karabay AZ, Aktan F, Sunguroğlu A, Buyukbingol Z. Methylsulfonylmethane modulates apoptosis of LPS/IFN-γ-activated RAW 264.7 macrophage-like cells by targeting p53, Bax, Bcl-2, cytochrome c and PARP proteins. Immunopharmacol Immunotoxicol. 2014;36(6):379–89. https://doi.org/10.3109/08923973.2014.956752.CrossRefPubMed

38.

Jani A, Ljubanovic D, Faubel S, Kim J, Mischak R, Edelstein CL. Caspase inhibition prevents the increase in caspase-3, -2, -8 and – 9 activity and apoptosis in the cold ischemic mouse kidney. Am J Transplant. 2004;4(8):1246–54. https://doi.org/10.1111/j.1600-6143.2004.00498.x.CrossRefPubMed

39.

Lin J, Qi W, Chen K, et al. Downregulating STAT1/caspase-3 signaling with fludarabine to alleviate progression in a rat model of steroid-induced avascular necrosis of the femoral head. J Biochem Mol Toxicol. 2019;33(4):e22265. https://doi.org/10.1002/jbt.22265.CrossRefPubMed

40.

Kuhrt D, Wojchowski DM. Emerging EPO and EPO receptor regulators and signal transducers. Blood. 2015;125(23):3536–41. https://doi.org/10.1182/blood-2014-11-575357.CrossRefPubMedPubMedCentral

41.

Haine L, Yegen CH, Marchant D, Richalet JP, Boncoeur E, Voituron N. Cytoprotective effects of erythropoietin: what about the lung? Biomed Pharmacother. 2021;139:111547. https://doi.org/10.1016/j.biopha.2021.111547.CrossRefPubMed

42.

Macdougall IC, Cooper AC. Erythropoietin resistance: the role of inflammation and pro-inflammatory cytokines. Nephrol Dial Transplant. 2002;17:39–43. https://doi.org/10.1093/ndt/17.suppl_11.39.CrossRefPubMed

43.

Gu Q, Chen M, Zhang Y, Huang Y, Yang H, Shi Q. Haem oxygenase-1 induction prevents glucocorticoid-induced osteoblast apoptosis through activation of extracellular signal-regulated kinase1/2 signalling pathway. J Orthop Translat. 2019;19:29–37. https://doi.org/10.1016/j.jot.2019.04.003.CrossRefPubMedPubMedCentral

44.

Leyland-Jones B, O’shaughnessy JA. Erythropoietin as a critical component of Breast cancer therapy: survival, synergistic, and cognitive applications. Semin Oncol. 2003;30(5 Suppl 16):174–84. https://doi.org/10.1053/j.seminoncol.2003.08.020.CrossRefPubMed

45.

Bullard AJ, Govewalla P, Yellon DM. Erythropoietin protects the myocardium against reperfusion injury in vitro and in vivo. Basic Res Cardiol. 2005;100(5):397–403. https://doi.org/10.1007/s00395-005-0537-4.CrossRefPubMed

46.

Jiang LY, Yu X, Pang QJ. Research in the precaution of recombinant human erythropoietin to steroid-induced osteonecrosis of the rat femoral head. J Int Med Res. 2017;45(4):1324–31. https://doi.org/10.1177/0300060517707076.CrossRefPubMedPubMedCentral

47.

Kumar A, Commane M, Flickinger TW, Horvath CM, Stark GR. Defective TNF-alpha-induced apoptosis in STAT1-null cells due to low constitutive levels of caspases. Science. 1997;278(5343):1630–2. https://doi.org/10.1126/science.278.5343.1630.CrossRefPubMed

48.

Sironi JJ, Ouchi T. STAT1-induced apoptosis is mediated by caspases 2, 3, and 7. J Biol Chem. 2004;279(6):4066–74. https://doi.org/10.1074/jbc.M307774200.CrossRefPubMed

49.

Thomas M, Finnegan CE, Rogers KMA, et al. STAT1: a modulator of chemotherapy-induced apoptosis. Cancer Res. 2004;64(22):8357–64. https://doi.org/10.1158/0008-5472.CAN-04-1864.CrossRefPubMed

50.

Xing YW, Liu KZ. Azithromycin inhibited oxidative stress and apoptosis of high glucose-induced podocytes by inhibiting STAT1 pathway. Drug Dev Res. 2021;82(7):990–8. https://doi.org/10.1002/ddr.21801.CrossRefPubMed

51.

Du X, Wang X, Cui K, et al. Tanshinone IIA and Astragaloside IV inhibit miR-223/JAK2/STAT1 signalling pathway to Alleviate Lipopolysaccharide-Induced damage in Nucleus Pulposus cells. Dis Markers. 2021;2021:6554480. https://doi.org/10.1155/2021/6554480.CrossRefPubMedPubMedCentral

Title: Erythropoietin suppresses osteoblast apoptosis and ameliorates steroid-induced necrosis of the femoral head in rats by inhibition of STAT1-caspase 3 signaling pathway
Authors: Tingwen Cai
Siyuan Chen
Chenghu Wu
Chao Lou
Weidan Wang
Chihao Lin
Hongyi Jiang
Xinxian Xu
Publication date: 01-12-2023
Publisher: BioMed Central
Keywords: Erythropoietin
Erythropoietin
Epoetin alfa
Dexamethasone
Published in: BMC Musculoskeletal Disorders / Issue 1/2023
Electronic ISSN: 1471-2474
DOI: https://doi.org/10.1186/s12891-023-07028-y

At a glance: The STEP trials

Springer Medicine

Erythropoietin suppresses osteoblast apoptosis and ameliorates steroid-induced necrosis of the femoral head in rats by inhibition of STAT1-caspase 3 signaling pathway

Abstract

Background

Objective

Method

Results

Conclusion

At a glance: The STEP trials

Springer Medicine

Abstract

Background

Objective

Method

Results

Conclusion

Please log in to get access to this content

Other articles of this Issue 1/2023

HTLV-1-associated myelopathy/tropical spastic paraplegia with sporadic late-onset nemaline myopathy: a case report

Correction: Assessments of early patellofemoral joint osteoarthritis features after anterior cruciate ligament reconstruction: a cross-sectional study

Total hip arthroplasty versus hemiarthroplasty in the treatment of active elderly patients over 75 years with displaced femoral neck fractures: a retrospective study

A first step towards a framework for interventions for individual working practice to prevent work-related musculoskeletal disorders: a scoping review

Pseudoarthrosis after anterior cervical discectomy and fusion: rate of occult infections and outcome of anterior revision surgery

The impact of neck pain on gait health: a systematic review and meta-analysis