Research of simulated microgravity regulate MC3T3-E1 cells differentiation through the nuclear factor-kappa B signaling pathway_Journal of Biomedical Engineering

Authors：

HAN Biao , ZHANG Yang , LI Hao , WEI Shuping , LI Ruixin ,  ZHANG Xizheng

Institute of Medical Service and Technology, Academy of System Engineering, Academy of Military Sciences, Tianjin 300161, P.R.China;

Corresponding?author：

ZHANG Xizheng, Email: z56787@sohu.com

Keywords：

simulated microgravity; osteoblast; nuclear factor-kappa B signaling pathway; bone morphogenetic proteins; osteoblast differentiation

DOI：

10.7507/1001-5515.201801019

Video：

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Abstract Full text Figures/Tables Video References Cited by

In this study, we aim to investigat the effect of microgravity on osteoblast differentiation in osteoblast-like cells (MC3T3-E1). In addition, we explored the response mechanism of nuclear factor-kappa B (NF-κB) signaling pathway to " zero-g” in MC3T3-E1 cells under the simulated microgravity conditions. MC3T3-E1 were cultured in conventional (CON) and simulated microgravity (SMG), respectively. Then, the expression of the related osteoblastic genes and the specific molecules in NF-κB signaling pathway were measured. The results showed that the mRNA and protein levels of alkaline phosphatase (ALP), osteocalcin (OCN) and type Ⅰ collagen (CoL-Ⅰ) were dramatically decreased under the simulated microgravity. Meanwhile, the NF-κB inhibitor α (IκB-α) protein level was decreased and the expressions of phosphorylation of IκB-α (p-IκB-α), p65 and phosphorylation of p65 (p-p65) were significantly up-regulated in SMG group. In addition, the IL-6 content in SMG group was increased compared to CON. These results indicated that simulated microgravity could activate the NF-κB pathway to regulate MC3T3-E1 cells differentiation.

Citation： HAN Biao, ZHANG Yang, LI Hao, WEI Shuping, LI Ruixin, ZHANG Xizheng. Research of simulated microgravity regulate MC3T3-E1 cells differentiation through the nuclear factor-kappa B signaling pathway. Journal of Biomedical Engineering, 2019, 36(3): 421-427. doi: 10.7507/1001-5515.201801019 Copy

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19.	Nakamura H, Aoki K, Masuda W, et al. Disruption of NF-kB1 prevents bone loss caused by mechanical unloading. J Bone Miner Res, 2013, 28(6): 1457-1467.
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25.	Csiszar A, Smith K E, Koller A, et al. Regulation of bone morphogenetic protein-2 expression in endothelial cells: role of nuclear factor-kappaB activation by tumornecrosis factor-alpha, H₂O₂, and high intravascular pressure. Circulation, 2005, 111(18): 2364-2372.
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1. Moustafa A, Sugiyama T, Saxon L K, et al. The mouse fibula as a suitable bone for the study of functional adaptation to mechanical loading. Bone, 2009, 44: 930-935.
2. Lambers F M, Schulte F A, Kuhn G A, et al. Mouse tail vertebrae adapt to cyclic mechanical loading by increasing bone formation rate and decreasing bone resorption rate as shown by time-lapsed in vivo imaging of dynamic bone morphometry. Bone, 2011, 49(6): 1340-1350.
3. White R J, Averner M. Humans in space. Nature, 2001, 409(6823): 1115-1118.
4. Allen D L, Bandstra E R, Harrison B C, et al. Effects of spaceflight on murine skeletal muscle gene expression. J Appl Physiol, 2009, 106(2): 582-595.
5. Hatton D C, Yue Q, Dierickx J, et al. Calcium metabolism and cardiovascular function after spaceflight. J Appl Physiol, 2002, 92(1): 3-12.
6. Crucian B, Sams C. Immune system dysregulation during spaceflight: clinical risk for exploration-class missions. J Leukoc Biol, 2009, 86(5): 1017-1018.
7. Ben-Neriah Y, Karin M. Inflammation meets cancer, with NF-kappa B as the matchmaker. Nat Immunol, 2011, 12(8): 715-723.
8. Hayden M S, Ghosh S. NF-kB, the first quarter-century: remarkable progress and outstanding questions. Genes Dev, 2012, 26(3): 203-234.
9. Chang Jia, Wang Zhuo, Tang E, et al. Inhibition of osteoblastic bone formation by nuclear factor-kappaB. Nat Med, 2009, 15(6): 682-689.
10. Yamazaki M, Fukushima H, Shin M, et al. TNFα represses BMP signaling by interfering with the DNA binding of Smads through the activation of NF-kB. J Biol Chem, 2004, 284(51): 35987-35995.
11. Alles N, Soysa N S, Hayashi J, et al. Suppression of NF-kB increases bone formation and ameliorates osteopenia in ovariectomized mice. Endocrinology, 2010, 151(10): 4626-4634.
12. Wang L, Li J, Zhang X, et al. Involvement of p38MAPK/NF-kappaB signaling pathways in osteoblasts differentiation in response to mechanical stretch. Ann Biomed Eng, 2012, 40(9): 1884-1894.
13. Livak K J, Schmittgen T D. Analysis of relative gene expression data using real-time quantitative PCR and the $ 2^ - \Delta \Delta \rmC_\rmT$ method. Methods, 2001, 25(4): 402-408.
14. Wang Qiangsong, Zhang Xinchang, Li Ruixin, et al. A comparative study of mechanical strain, icariin and combination stimulations on improving osteoinductive potential via NF-kappaB activation in osteoblast-like cells. Biomed Eng Online, 2015, 14(1): 46.
15. Zhao J. Postdoc position at HKUST on computational mechanics/geomechanics. Oral Dis, 2012, 19(1): 59-64.
16. Frost H M. Bone " mass” and the " mechanostat”: a proposal. Anat Rec, 1987, 219(1): 1-9.
17. Zhang Xinchang, Wang Qiangsong, Wan Zongming, et al. CKIP-1 knockout offsets osteoporosis induced by simulated microgravity. Prog Biophys Mol Biol, 2016, 122(2, SI): 140-148.
18. 韓標, 張新昌, 李昊, 等. 不同強度周期性動態力學載荷對微重力下骨質疏松影響. 醫用生物力學, 2017, 32(1): 83-87.
19. Nakamura H, Aoki K, Masuda W, et al. Disruption of NF-kB1 prevents bone loss caused by mechanical unloading. J Bone Miner Res, 2013, 28(6): 1457-1467.
20. Meyers V E, Zayzafoon M, Douglas J T, et al. RhoA and cytoskeletal disruption mediate reduced osteoblastogenesis and enhanced adipogenesis of human mesenchymal stem cells in modeled microgravity. J Bone Miner Res, 2005, 20(10): 1858-1866.
21. Plett P A, Abonour R, Frankovitz S M, et al. Impact of modeled microgravity on migration, differentiation, and cell cycle control of primitive human hematopoietic progenitor cells. Exp Hematol, 2004, 32(8): 773-781.
22. Zwart S R, Pierson D, Mehta S, et al. Capacity of omega-3 fatty acids or eicosapentaenoic acid to counteract weightlessness-induced bone loss by inhibiting NF-kappa B activation: from cells to bed rest to astronauts. J Bone Miner Res, 2010, 25(5): 1049-1057.
23. Huang Xiaofeng, Chai Yang. TGF-β signalling and tooth development. Chin J Dent Res, 2010, 13(1): 7-15.
24. Khanal A, Yoshioka I, Tominaga K, et al. The BMP signaling and its Smads in mandibular distraction osteogenesis. Oral Dis, 2008, 14(4): 347-355.
25. Csiszar A, Smith K E, Koller A, et al. Regulation of bone morphogenetic protein-2 expression in endothelial cells: role of nuclear factor-kappaB activation by tumornecrosis factor-alpha, H₂O₂, and high intravascular pressure. Circulation, 2005, 111(18): 2364-2372.
26. Cazzaniga A, Castiglioni S, Maier J A M. Conditioned media from microvascular endothelial cells cultured in simulated microgravity inhibit osteoblast activity. Biomed Res Int, 2014, 2014: 857934.

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Research of simulated microgravity regulate MC3T3-E1 cells differentiation through the nuclear factor-kappa B signaling pathway

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