DEC1 deficiency protects against bone loss induced by ovariectomy by inhibiting inflammation

Abstract

Studies have shown that differentiated embryo-chondrocyte expressed gene 1 (DEC1) promotes osteoblast osteogenesis. To investigate the role of DEC1 in postmenopausal osteoporosis, we used the two genotypes of mice (Dec1^+/+ and Dec1^−/−) to establish an ovariectomy model and found that the bone loss was significantly lower in Dec1^−/− ovariectomy mice than in Dec1^+/+ ovariectomy mice. The expression levels of RUNX2 and OSX were significantly increased in Dec1^−/− ovariectomy mice, compared with Dec1^+/+ ovariectomy mice; however, the expression levels of NFATc1, c-Fos, CTSK, and RANKL/OPG ratio were significantly decreased in Dec1^−/− ovariectomy mice, compared with those in Dec1^+/+ ovariectomy mice. Likewise, DEC1 deficiency also suppressed the expression levels of IL-6 and IL-1β. Further results showed that the mRNA expression levels of Runx2, Osx, and Alp were significantly increased in bone marrow mesenchymal stem cells of Dec1^−/− ovariectomy mice, compared with those of Dec1^+/+ ovariectomy mice. Moreover, the mRNA levels of Il1b, Il6, Tnfa, and Ifng were significantly increased in bone marrow-derived macrophages (BMMs) of Dec1^+/+ovariectomy mice, compared with those of Dec1^+/+ sham mice, but not in Dec1^−/− ovariectomy BMMs, when compared with those in Dec1^−/− sham BMMs. Additionally, the expression levels of p-IκBα and p-P65 were significantly increased in Dec1^+/+ ovariectomy BMMs, compared with those in Dec1^+/+ sham BMMs, but did not increase in Dec1^−/− ovariectomy BMMs, compared with those in Dec1^−/− sham BMMs. Taken together, DEC1 deficiency inhibited the NF-κB pathway induced by ovariectomy, thereby decreasing cytokines and subsequently inhibiting the decrease of osteogenesis and the increase of osteoclastogenesis caused by ovariectomy. The findings may provide a novel understanding of postmenopausal osteoporosis development, and offer potential avenues for the disease intervention.

Keywords

DEC1 / postmenopausal osteoporosis / estrogen / osteoblasts / osteoclasts

Fundings

This work was supported by the National Natural Science Foundation of China (Grant Nos. 82073934 and 81872937 to J.Y.), and the Special Foundation for Clinical Science and Technology of Jiangsu Province (Grant No. JX10114120).

Acknowledgments

None.

References

[1] Kanis JA, Johnell O, Oden A, et al. Long-term risk of osteoporotic fracture in Malmö[J]. Osteoporos Int, 2000, 11(8): 669-674. doi: 10.1007/s001980070064

[2] Reid IR. A broader strategy for osteoporosis interventions[J]. Nat Rev Endocrinol, 2020, 16(6): 333-339. doi: 10.1038/s41574-020-0339-7

[3] Feng X, McDonald JM. Disorders of bone remodeling[J]. Annu Rev Pathol Mech Dis, 2011, 6: 121-145. doi: 10.1146/annurev-pathol-011110-130203

[4] Capulli M, Paone R, Rucci N. Osteoblast and osteocyte: Games without frontiers[J]. Arch Biochem Biophys, 2014, 561: 3-12. doi: 10.1016/j.abb.2014.05.003

[5] Yousefzadeh N, Kashfi K, Jeddi S, et al. Ovariectomized rat model of osteoporosis: A practical guide[J]. Excli J, 2020, 19: 89-107. https://pubmed.ncbi.nlm.nih.gov/32038119/

[6] Komori T. Regulation of proliferation, differentiation and functions of osteoblasts by runx2[J]. Int J Mol Sci, 2019, 20(7): 1694. doi: 10.3390/ijms20071694

[7] Komori T. Runx2, an inducer of osteoblast and chondrocyte differentiation[J]. Histochem Cell Biol, 2018, 149(4): 313-323. doi: 10.1007/s00418-018-1640-6

[8] Ru J, Wang Y. Osteocyte apoptosis: The roles and key molecular mechanisms in resorption-related bone diseases[J]. Cell Death Dis, 2020, 11(10): 846. doi: 10.1038/s41419-020-03059-8

[9] Yang C, Tao H, Zhang H, et al. TET2 regulates osteoclastogenesis by modulating autophagy in OVX-induced bone loss[J]. Autophagy, 2022, 18(12): 2817-2829. doi: 10.1080/15548627.2022.2048432

[10] Han S, Kim JH, Jo EH, et al. Eleutherococcus sessiliflorus inhibits receptor activator of nuclear factor kappa-B ligand (RANKL)-induced osteoclast differentiation and prevents ovariectomy (OVX)-induced bone loss[J]. Molecules, 2021, 26(7): 1886. doi: 10.3390/molecules26071886

[11] Hong G, Zhou L, Han X, et al. Asiatic acid inhibits OVX-induced osteoporosis and osteoclastogenesis via regulating RANKL-Mediated NF-κb and nfatc1 signaling pathways[J]. Front Pharmacol, 2020, 11: 331. doi: 10.3389/fphar.2020.00331

[12] Kim JH, Kim N. Regulation of NFATc1 in osteoclast differentiation[J]. J Bone Metab, 2014, 21(4): 233-241. doi: 10.11005/jbm.2014.21.4.233

[13] Kim WS, Kim HJ, Lee ZH, et al. Apolipoprotein E inhibits osteoclast differentiation via regulation of c-Fos, NFATc1 and NF-κB[J]. Exp Cell Res, 2013, 319(4): 436-446. doi: 10.1016/j.yexcr.2012.12.004

[14] Teitelbaum SL. Bone resorption by osteoclasts[J]. Science, 2000, 289(5484): 1504-1508. doi: 10.1126/science.289.5484.1504

[15] Kong YY, Yoshida H, Sarosi I, et al. OPGL is a key regulator of osteoclastogenesis, lymphocyte development and lymph-node organogenesis[J]. Nature, 1999, 397(6717): 315-323. doi: 10.1038/16852

[16] Cheng C, Chen L, Chen K. Osteoporosis due to hormone imbalance: An overview of the effects of estrogen deficiency and glucocorticoid overuse on bone turnover[J]. Int J Mol Sci, 2022, 23(3): 1376. doi: 10.3390/ijms23031376

[17] Pfeilschifter J, Köditz R, Pfohl M, et al. Changes in proinflammatory cytokine activity after menopause[J]. Endocr Rev, 2002, 23(1): 90-119. doi: 10.1210/edrv.23.1.0456

[18] Yamada K, Miyamoto K. Basic helix-loop-helix transcription factors, BHLHB2 and BHLHB3; their gene expressions are regulated by multiple extracellular stimuli[J]. Front Biosci, 2005, 10(3): 3151-3171. doi: 10.2741/1772

[19] Jia Y, Xiao D, Ma X, et al. Differentiated embryonic chondrocyte-expressed gene 1 is associated with hypoxia-inducible factor 1α and Ki67 in human gastric cancer[J]. Diagn Pathol, 2013, 8(1): 37. doi: 10.1186/1746-1596-8-37

[20] Li S, Peng D, Yin Z, et al. Effect of DEC1 on the proliferation, adhesion, invasion and epithelial-mesenchymal transition of osteosarcoma cells[J]. Exp Ther Med, 2020, 19(3): 2360-2366. doi: 10.3892/etm.2020.8459

[21] Shan E, Huo Y, Wang H, et al. Differentiated embryonic chondrocyte expressed gene-1 (DEC1) enhances the development of colorectal cancer with an involvement of the STAT3 signaling[J]. Neoplasia, 2022, 27: 100783. doi: 10.1016/j.neo.2022.100783

[22] Yuan N, Liu Y, Zhang D. Role of differentiated embryo-chondrocyte expressed gene 1 (DEC1) in immunity[J]. Int Immunopharmacol, 2022, 102: 107892. doi: 10.1016/j.intimp.2021.107892

[23] Shen M, Yoshida E, Yan W, et al. Basic helix-loop-helix protein DEC1 promotes chondrocyte differentiation at the early and terminal stages[J]. J Biol Chem, 2002, 277(51): 50112-50120. doi: 10.1074/jbc.M206771200

[24] Hu J, Mao Z, He S, et al. Icariin protects against glucocorticoid induced osteoporosis, increases the expression of the bone enhancer DEC1 and modulates the PI3K/Akt/GSK3β/β-catenin integrated signaling pathway[J]. Biochem Pharmacol, 2017, 136: 109-121. doi: 10.1016/j.bcp.2017.04.010

[25] Hu J, Mao Z, Shang W, et al. Down regulation of differentiated embryo-chondrocyte expressed gene 1 is related to the decrease of osteogenic capacity[J]. Curr Drug Targets, 2014, 15(4): 432-441. doi: 10.2174/1389450114666140102133719

[26] Ali D, Figeac F, Caci A, et al. High-fat diet-induced obesity augments the deleterious effects of estrogen deficiency on bone: Evidence from ovariectomized mice[J]. Aging Cell, 2022, 21(12): e13726. doi: 10.1111/acel.13726

[27] Li Y, Zhuang Q, Tao L, et al. Urolithin B suppressed osteoclast activation and reduced bone loss of osteoporosis via inhibiting ERK/NF-κB pathway[J]. Cell Prolif, 2022, 55(10): e13291. doi: 10.1111/cpr.13291

[28] Weitzmann MN, Pacifici R. Estrogen deficiency and bone loss: an inflammatory tale[J]. J Clin Invest, 2006, 116(5): 1186-1194. doi: 10.1172/JCI28550

[29] Weitzmann MN. The role of inflammatory cytokines, the RANKL/OPG axis, and the immunoskeletal interface in physiological bone turnover and osteoporosis[J]. Scientifica (Cairo), 2013, 2013: 125705. doi: 10.1155/2013/125705

[30] Pittenger MF, Mackay AM, Beck SC, et al. Multilineage potential of adult human mesenchymal stem cells[J]. Science, 1999, 284(5411): 143-147. doi: 10.1126/science.284.5411.143

[31] Zhuang X, Hu G. In vitro osteoclastogenesis assays using primary mouse bone marrow cells[J]. Bio Protoc, 2018, 8(11): e2875. doi: 10.21769/BioProtoc.2875

[32] Kang IS, Kim C. NADPH oxidase gp91^phox contributes to RANKL-induced osteoclast differentiation by upregulating NFATc1[J]. Sci Rep, 2016, 6: 38014. doi: 10.1038/srep38014

[33] Wu Y, Humphrey MB, Nakamura MC. Osteoclasts—the innate immune cells of the bone[J]. Autoimmunity, 2008, 41(3): 183-194. doi: 10.1080/08916930701693180

[34] Weischenfeldt J, Porse B. Bone marrow-derived macrophages (BMM): isolation and applications[J]. CSH Protoc, 2008, 2008: pdb. prot5080.

[35] Medzhitov R. Origin and physiological roles of inflammation[J]. Nature, 2008, 454(7203): 428-435. doi: 10.1038/nature07201

[36] Salmon AJ, Shavkunov AS, Miao Q, et al. BHLHE40 regulates the T-cell effector function required for tumor microenvironment remodeling and immune checkpoint therapy efficacy[J]. Cancer Immunol Res, 2022, 10(5): 597-611. doi: 10.1158/2326-6066.CIR-21-0129

[37] Li X, Zhang X, Hou X, et al. Obstructive sleep apnea-increased DEC1 regulates systemic inflammation and oxidative stress that promotes development of pulmonary arterial hypertension[J]. Apoptosis, 2023, 28(3-4): 432-446. doi: 10.1007/s10495-022-01797-y

[38] Amarasekara DS, Kim S, Rho J. Regulation of osteoblast differentiation by cytokine networks[J]. Int J Mol Sci, 2021, 22(6): 2851. doi: 10.3390/ijms22062851

[39] Xu J, Yu LX, Liu F, et al. The effect of cytokines on osteoblasts and osteoclasts in bone remodeling in osteoporosis: A review[J]. Front Immunol, 2023, 14: 1222129. doi: 10.3389/fimmu.2023.1222129

[40] Yokoe S, Hasuike A, Watanabe N, et al. Epstein-barr virus promotes the production of inflammatory cytokines in gingival fibroblasts and RANKL-induced osteoclast differentiation in RAW264.7 cells[J]. Int J Mol Sci, 2022, 23(2): 809. doi: 10.3390/ijms23020809

[41] Goettems-Fiorin PB, Costa-Beber LC, Dos Santos JB, et al. Ovariectomy predisposes female rats to fine particulate matter exposure's effects by altering metabolic, oxidative, pro-inflammatory, and heat-shock protein levels[J]. Environ Sci Pollut Res Int, 2019, 26(20): 20581-20594. doi: 10.1007/s11356-019-05383-9

[42] Mansoori MN, Raghuvanshi A, Shukla P, et al. Medicarpin prevents arthritis in post-menopausal conditions by arresting the expansion of TH17 cells and pro-inflammatory cytokines[J]. Int Immunopharmacol, 2020, 82: 106299. doi: 10.1016/j.intimp.2020.106299

[43] Rodríguez-Berriguete G, Prieto A, Fraile B, et al. Relationship between IL-6/ERK and NF-κB: A study in normal and pathological human prostate gland[J]. Eur Cytokine Netw, 2010, 21(4): 241-250. doi: 10.1684/ecn.2010.0211

[44] Abohassan M, Al Shahrani M, Alshahrani MY, et al. FNF-12, a novel benzylidene-chromanone derivative, attenuates inflammatory response in in vitro and in vivo asthma models mediated by M2-related Th2 cytokines via MAPK and NF-kB signaling[J]. Pharmacol Rep, 2022, 74(1): 96-110. doi: 10.1007/s43440-021-00325-0

[45] Sun B, Wang B, Xu M. Esculetin inhibits histamine-induced expression of inflammatory cytokines and mucin in nasal epithelial cells[J]. Clin Exp Pharmacol Physiol, 2019, 46(9): 821-827. doi: 10.1111/1440-1681.13128

[46] Jimi E. The role of BMP signaling and NF-κB signaling on osteoblastic differentiation, cancer development, and vascular diseases—Is the activation of NF-κB a friend or foe of BMP function?[J]. Vitam Horm, 2015, 99: 145-170. doi: 10.1016/bs.vh.2015.05.002