Effect of Osteopontin on Polarization of Macrophage in Vitro.

doi:10.13701/j.cnki.kqyxyj.2019.05.020

Abstract

Abstract: Objective: To study the effect of exogenous osteopontin (OPN) on the polarization of macrophage in vitro. Methods: Mouse RAW264.7 macrophages were cultured in vitro, and divided into groups as follows: (1) OPN dose-dependent experiment: the experimental group were treated with 0.1,0.5,1.0 mg/L OPN respectively in vitro; (2) M1 polarization experiment: set control group, LPS group, OPN group, LPS+OPN group; (3) M2 polarization experiment: there were control group, IL-4 group, OPN group, and IL-4+OPN group. After stimulated for 24 hours, quantitative real-time PCR (qRT-PCR) was used to measure mRNA levels of M1-type genes inducible nitric oxide synthase (iNOS), tumor necrosis factor-α (TNF-α), interleukin-1β (IL-1β), and M2-type genes CD206, Arginase-1 (Arg-1), and interleukin-10 (IL-10). Western blot was used to assay iNOS and Arg-1 protein. The expression of iNOS and CD206 was detected by immunofluorescence assay (IFA). Results: mRNA levels of M1 and M2-type genes both increased significantly in a dose-dependent manner on OPN group (P<0.05). The expression of M1-type genes was the highest on LPS+OPN group, while expression of M2-type genes was the highest on IL-4+OPN group (P<0.05). The Western blot and IFA results were consistent with qRT-PCR results. Conclusion: OPN may induce M1 or M2 polarization of macrophage, and has synergetic effect with LPS and IL-4, as a pleiotropic cytokine.

Key words: Osteopontin, Macrophage, M1 polarization, M2 polarization

FENG Li-hua, WANG Min, XIA Hai-bin, HUANG Kai-yao. Effect of Osteopontin on Polarization of Macrophage in Vitro.[J]. Journal of Oral Science Research, 2019, 35(5): 497-501.

References

[1] Kzhyshkowska J, Gudima A, Riabov V, et al. Macrophage responses to implants: prospects for personalized medicine [J]. J Leukoc Biol, 2015, 98(6):953-962.
[2] Martinez FO, Gordon S. The M1 and M2 paradigm of macrophage activation: time for reassessment [J]. F1000Prime Rep, 2014, 6:13.
[3] Ivashkiv LB. Epigenetic regulation of macrophage polarization and function [J]. Trends Immunol, 2013, 34(5):216-223.
[4] Murray PJ, Allen JE, Biswas SK, et al. Macrophage activation and polarization: nomenclature and experimental guidelines [J]. Immunity, 2014, 41(1):14-20.
[5] Morris AH, Kyriakides TR. Matricellular proteins and biomaterials [J]. Matrix Biol, 2014, 37:183-191.
[6] 符策广,刘洁,夏海斌.骨桥蛋白的生物学特性及在骨改建中的作用[J].临床口腔医学杂志,2012,28(8):506-508.
[7] Icer MA, Gezmen-Karadag M. The multiple functions and mechanisms of osteopontin [J]. Clin Biochem,2018,59:17-24.
[8] Mckee MD, Pedraza CE, Kaartinen MT. Osteopontin and wound healing in bone [J]. Cells Tissues Organs,2011,194(2-4):313-319.
[9] Makishi S, Saito K, Ohshima H. Osteopontin-deficiency disturbs direct osteogenesis in the process of achieving osseointegration following immediate placement of endosseous implants [J]. Clin Implant Dent Relat Res, 2017, 19(3):496-504.
[10] Singh A, Gill G, Kaur H, et al. Role of osteopontin in bone remodeling and orthodontic tooth movement: a review [J]. Prog Orthod, 2018, 19(1):18.
[11] 史欣,张鹏飞,袁梦桐,等.骨桥蛋白对人牙髓干细胞成骨分化的旁分泌作用[J].口腔医学研究,2014,30(11):1058-1061.
[12] Parisi L, Gini E, Baci D, et al. Macrophage polarization in chronic inflammatory diseases: killers or builders [J]. J Immunol Res, 2018, 2018:8917804.
[13] Heilmann K, Hoffmann U, Witte E, et al. Osteopontin as two-sided mediator of intestinal inflammation [J]. J Cell Mol Med, 2009, 13(6):1162-1174.
[14] Yang H, Ye X, Zhang X, et al. Intracellular osteopontin negatively regulates toll-like receptor 4-mediated inflammatory response via regulating GSK3β and 4EBP1 phosphorylation [J]. Cytokine, 2018, 108:89-95.
[15] Many GM, Yokosaki Y, Uaesoontrachoon K, et al. OPN-a induces muscle inflammation by increasing recruitment and activation of pro-inflammatory macrophages [J]. Exp Physiol, 2016, 101(10):1285-1300.
[16] Arango Duque G, Descoteaux A. Macrophage cytokines: involvement in immunity and infectious diseases [J]. Front Immunol, 2014, 5:491.
[17] 李明政,肖宇,吴珍珍,等.巨噬细胞在材料诱导成骨中作用的研究进展[J].口腔医学研究,2017,33(10):1123-1126.
[18] Mosser DM, Edwards JP. Exploring the full spectrum of macrophage activation [J]. Nat Rev Immunol, 2008, 8(12):958-969.
[19] Brown BN, Ratner BD, Goodman SB, et al. Macrophage polarization: An opportunity for improved outcomes in biomaterials and regenerative medicine [J]. Biomaterials, 2012, 33(15):3792-3802.