Naringenin promotes SDF-1/CXCR4 signaling pathway in BMSCs osteogenic differentiation
Abstract
Introduction. Naringenin, a dihydro-flavonoid compound that shows chemotactic activity, may have a good application prospect in repairing bone tissue, but its specific mechanism in bone regeneration, especially the osteogenic differentiation of stem cells, needs for a further study. The aim of this study was to investigate the effect of naringenin on the osteogenic differentiation and its roles in the C-X-C chemokine receptor type 4/stromal cell-derived factor 1 (SDF-1/CXCR4) signal pathway of bone marrow-derived mesenchymal stem cells (BMSCs).
Material and methods. BMSCs were harvested from the femurs and tibias of 4-to-6-week-old male Sprague-Dawley rats. Cell Counting kit-8 assay was used to determine cytotoxicity of naringenin. Alkaline phosphatase (ALP) activity was measured in cell’s precipitates and alizarin-red staining was performed to determine the osteogenic differentiation capacity of the BMSCs. Real-time polymerase chain reaction, enzyme-linked immunosorbent assay and western blotting were adopted to determine the expression of genes and proteins.
Results. The cellular morphology was spindle-shaped, and arranged in radial and whorled patterns. The flow cytometric analysis have confirmed the presence of characteristic surface proteins in the harvested BMSCs. Different concentrations (0–200 μg/ml) of naringenin have no influence on the viability and proliferation rate of the BMSCs. The highest ALP activity was found at culture day 7 and 9 when the concentration of naringenin was 75 and 100 μg/ml. Positive red or dark red stained cells with mineralized nodules can be observed on day 14. The expression of ALP, Runt-related transcription factor 2, CXCR4 and SDF-1a at the gene and protein levels in naringenin-treated cells were significantly higher than those in the control cells. Moreover, AMD3100, an inhibitor of CXCR4, suppressed the expression of the studied genes and proteins.
Conclusions. Naringenin does not show toxic effect on BMSCs. Naringenin promotes the expression of the SDF-1a gene and protein via the SDF-1/CXCR4 signaling pathway. A better understanding of the mechanisms of naringenin action would be helpful for developing specific therapeutic strategies to improve bone regeneration after injuries.
Keywords: bone marrow-derived mesenchymal stem cellscell viabilitynaringeninalizarin redSDF-1CXCR4RT-qPCR
References
- Hokugo A, Saito T, Li A, et al. Stimulation of bone regeneration following the controlled release of water-insoluble oxysterol from biodegradable hydrogel. Biomaterials. 2014; 35(21): 5565–5571.
- Fong ELS, Chan CK, Goodman SB. Stem cell homing in musculoskeletal injury. Biomaterials. 2011; 32(2): 395–409.
- Sha Y, Lv Y, Xu Z, et al. MGF E peptide pretreatment improves the proliferation and osteogenic differentiation of BMSCs via MEK-ERK1/2 and PI3K-Akt pathway under severe hypoxia. Life Sci. 2017; 189: 52–62.
- Wu J, Zhang W, Ran Q, et al. The Differentiation Balance of Bone Marrow Mesenchymal Stem Cells Is Crucial to Hematopoiesis. Stem Cells Int. 2018; 2018: 1540148.
- Kawakami Y, Ii M, Matsumoto T, et al. SDF-1/CXCR4 axis in Tie2-lineage cells including endothelial progenitor cells contributes to bone fracture healing. J Bone Miner Res. 2015; 30(1): 95–105.
- Peyvandi AA, Roozbahany NA, Peyvandi H, et al. Critical role of SDF-1/CXCR4 signaling pathway in stem cell homing in the deafened rat cochlea after acoustic trauma. Neural Regen Res. 2018; 13(1): 154–160.
- Zhu W, Liang G, Huang Z, et al. Conditional inactivation of the CXCR4 receptor in osteoprecursors reduces postnatal bone formation due to impaired osteoblast development. J Biol Chem. 2011; 286(30): 26794–26805.
- Herberg S, Kondrikova G, Hussein KA, et al. Mesenchymal stem cell expression of stromal cell-derived factor-1β augments bone formation in a model of local regenerative therapy. J Orthop Res. 2015; 33(2): 174–184.
- Toupadakis CA, Wong A, Genetos DC, et al. Long-term administration of AMD3100, an antagonist of SDF-1/CXCR4 signaling, alters fracture repair. J Orthop Res. 2012; 30(11): 1853–1859.
- Liles W, Broxmeyer H, Rodger E, et al. Mobilization of hematopoietic progenitor cells in healthy volunteers by AMD3100, a CXCR4 antagonist. Blood. 2003; 102(8): 2728–2730.
- Joshi R, Kulkarni YA, Wairkar S. Pharmacokinetic, pharmacodynamic and formulations aspects of Naringenin: An update. Life Sci. 2018; 215: 43–56.
- Cavia-Saiz M, Busto MD, Pilar-Izquierdo MC, et al. Antioxidant properties, radical scavenging activity and biomolecule protection capacity of flavonoid naringenin and its glycoside naringin: a comparative study. J Sci Food Agric. 2010; 90(7): 1238–1244.
- Li YR, Chen DY, Chu CL, et al. Naringenin inhibits dendritic cell maturation and has therapeutic effects in a murine model of collagen-induced arthritis. J Nutr Biochem. 2015; 26(12): 1467–1478.
- Pinho-Ribeiro FA, Zarpelon AC, Fattori V, et al. Naringenin reduces inflammatory pain in mice. Neuropharmacology. 2016; 105: 508–519.
- Yi LT, Li CF, Zhan X, et al. Involvement of monoaminergic system in the antidepressant-like effect of the flavonoid naringenin in mice. Prog Neuropsychopharmacol Biol Psychiatry. 2010; 34(7): 1223–1228.
- Kaczmarczyk-Sedlak I, Wojnar W, Zych M, et al. EFFECT OF DIETARY FLAVONOID NARINGENIN ON BONES IN RATS WITH OVARIECTOMY-INDUCED OSTEOPOROSIS. Acta Pol Pharm. 2016; 73(4): 1073–1081.
- Swarnkar G, Sharan K, Siddiqui JA, et al. A naturally occurring naringenin derivative exerts potent bone anabolic effects by mimicking oestrogen action on osteoblasts. Br J Pharmacol. 2012; 165(5): 1526–1542.
- Wang X, Zhen L, Zhang Ge, et al. Osteogenic effects of flavonoid aglycones from an osteoprotective fraction of Drynaria fortunei--an in vitro efficacy study. Phytomedicine. 2011; 18(10): 868–872.
- Wang W, Wu C, Tian Bo, et al. The Inhibition of RANKL-Induced Osteoclastogenesis through the Suppression of p38 Signaling Pathway by Naringenin and Attenuation of Titanium-Particle-Induced Osteolysis. Int J Mol Sci. . 2014; 15(12): 21913–21934.
- Wang W, Li M, Luo M, et al. Naringenin inhibits osteoclastogenesis through modulation of helper T cells-secreted IL-4. J Cell Biochem. 2018; 119(2): 2084–2093.
- Azizi SA, Stokes D, Augelli BJ, et al. Engraftment and migration of human bone marrow stromal cells implanted in the brains of albino rats--similarities to astrocyte grafts. Proc Natl Acad Sci U S A. 1998; 95(7): 3908–3913.
- Yin Y, Li F, Li S, et al. TLR4 Influences Hepatitis B Virus Related Hepatocellular Carcinoma by Regulating the Wnt/β-Catenin Pathway. Cell Physiol Biochem. 2017; 42(2): 469–479.
- Zhang X, Du Yu, Ling J, et al. Dickkopf-related protein 3 negatively regulates the osteogenic differentiation of rat dental follicle cells. Mol Med Rep. 2017; 15(4): 1673–1681.
- Fang T, Wang Y, Ma Y, et al. A rapid LC/MS/MS quantitation assay for naringin and its two metabolites in rats plasma. J Pharm Biomed Anal. 2006; 40(2): 454–459.
- Hübel K, Liles W, Broxmeyer H, et al. Leukocytosis and Mobilization of CD34+ Hematopoietic Progenitor Cells by AMD3100, a CXCR4 Antagonist. Supportive Cancer Therapy. 2004; 1(3): 165–172.
- Toupadakis CA, Wong A, Genetos DC, et al. Long-term administration of AMD3100, an antagonist of SDF-1/CXCR4 signaling, alters fracture repair. J Orthop Res. 2012; 30(11): 1853–1859.
- Kitaori T, Ito H, Schwarz EM, et al. Stromal cell-derived factor 1/CXCR4 signaling is critical for the recruitment of mesenchymal stem cells to the fracture site during skeletal repair in a mouse model. Arthritis Rheum. 2009; 60(3): 813–823.
- Ming LG, Ge BF, Wang MG, et al. Comparison between 8-prenylnarigenin and narigenin concerning their activities on promotion of rat bone marrow stromal cells' osteogenic differentiation in vitro. Cell Prolif. 2012; 45(6): 508–515.
- Yu GY, Zheng GZ, Chang Bo, et al. Naringin Stimulates Osteogenic Differentiation of Rat Bone Marrow Stromal Cells via Activation of the Notch Signaling Pathway. Stem Cells Int. 2016; 2016: 7130653.
- Tamamura H, Kuroda M, Masuda M, et al. A comparative study of the solution structures of tachyplesin I and a novel anti-HIV synthetic peptide, T22 ([Tyr5,12, Lys7]-polyphemusin II), determined by nuclear magnetic resonance. Biochim Biophys Acta. 1993; 1163(2): 209–216.
- Nakashima H, Masuda M, Murakami T, et al. Anti-human immunodeficiency virus activity of a novel synthetic peptide, T22 ([Tyr-5,12, Lys-7]polyphemusin II): a possible inhibitor of virus-cell fusion. Antimicrob Agents Chemother. 1992; 36(6): 1249–1255.
- Kitaori T, Ito H, Schwarz EM, et al. Stromal cell-derived factor 1/CXCR4 signaling is critical for the recruitment of mesenchymal stem cells to the fracture site during skeletal repair in a mouse model. Arthritis Rheum. 2009; 60(3): 813–823.
- Yang F, Xue F, Guan J, et al. Stromal-Cell-Derived Factor (SDF) 1-Alpha Overexpression Promotes Bone Regeneration by Osteogenesis and Angiogenesis in Osteonecrosis of the Femoral Head. Cell Physiol Biochem. 2018; 46(6): 2561–2575.
- Wang S, Zhou C, Zheng H, et al. Chondrogenic progenitor cells promote vascular endothelial growth factor expression through stromal-derived factor-1. Osteoarthritis Cartilage. 2017; 25(5): 742–749.
- Chen HT, Tsou HK, Hsu CJ, et al. Stromal cell-derived factor-1/CXCR4 promotes IL-6 production in human synovial fibroblasts. J Cell Biochem. 2011; 112(4): 1219–1227.
- Wang H, Li C, Li J, et al. Naringin enhances osteogenic differentiation through the activation of ERK signaling in human bone marrow mesenchymal stem cells. Iran J Basic Med Sci. 2017; 20(4): 408–414.
- Wu JB, Fong YC, Tsai HY, et al. Naringin-induced bone morphogenetic protein-2 expression via PI3K, Akt, c-Fos/c-Jun and AP-1 pathway in osteoblasts. Eur J Pharmacol. 2008; 588(2-3): 333–341.
- Hernández-Aquino E, Muriel P. Beneficial effects of naringenin in liver diseases: Molecular mechanisms. World J Gastroenterol. 2018; 24(16): 1679–1707.
