Galangin inhibits lipopolysaccharide-induced inflammation and stimulates osteogenic differentiation of bone marrow mesenchymal stem cells via regulation of AKT/mTOR signaling

Main Article Content

Xiaoting Wang
Xinglei Xiao

Keywords

AKT/mTOR, bone marrow mesenchymal stem cells, galangin, inflammation, lipopolysaccharides, osteogenic differentiation

Abstract

Background: Bone marrow mesenchymal stem cells (BMSCs), with the abilities of multidirectional differentiation and self-renewal, have been widely used in bone repair and regeneration of inflammation-stimulated oral diseases. Galangin is a flavonoid isolated from Alpinia officinarum, exerts anti-obesity, antitumor, and anti-inflammation pharmacological effects. The roles of galangin in lipopolysaccharide-induced inflammation and osteogenic differentiation of BMSCs were investigated.


Methods: BMSCs were isolated from rat bone marrow and identified by flow cytometry. The isolated BMSCs were treated with 1 μg/mL lipopolysaccharides or cotreated with lipopolysaccharides and different concentrations of galangin. Cell viability and apoptosis were detected by MTT (tetrazolium component) and flow cytometry. ELISA was used to detect inflammation. Alizarin red staining was used to investigate osteogenic differentiation.


Results: The rat BMSCs showed negative rate of CD34, and positive rate of CD29 and CD44. Lipopolysaccharides treatment reduced cell viability of BMSCs, and promoted the cell apoptosis. Incubation with galangin enhanced cell viability of lipopolysaccharide-stimulated BMSCs, and suppressed the cell apoptosis. Galangin decreased levels of TNF-α, IL-1β, and IL-6 in lipopolysaccharide-stimulated BMSCs through down-regulation of NF-κB phosphorylation (p-NF-κB). Galangin up-regulated expression of osteo-specific proteins, collagen type I alpha 1 (COL1A1), osteopontin (OPN), and runt-related transcription factor 2 (RUNX2), to promote the osteogenic differentiation of lipopolysaccharide-stimulated BMSCs. Protein expression of p-AKT and p-mTOR in lipopolysaccharide-stimulated BMSCs were increased by galangin treatment.


Conclusion: Galangin exerted an anti-inflammatory effect against lipopolysaccharide- stimulated BMSCs and promoted osteogenic differentiation through the activation of AKT/ mTOR signaling.

Abstract 132 | PDF Downloads 65 HTML Downloads 9 XML Downloads 1

References

1. Ramenzoni LL, Lehner MP, Kaufmann ME, Wiedemeier D, Attin T, Schmidlin PR. Oral diagnostic methods for the detection of periodontal disease. Diagnostics. 2021;11(3):571. 10.3390/diagnostics11030571

2. Shiba T, Watanabe T, Komatsu K, Koyanagi T, Nemoto T, Ohsugi Y, et al. Non-surgical treatment for periodontitis and peri-implantitis: longitudinal clinical and bacteriological findings-A case report with a 7-year follow-up evaluation. SAGE Open Med Case Rep. 2021;9:2050313X211029154–2050313X. 10.1177/2050313X211029154

3. Wennström JL, Tomasi C, Bertelle A, Dellasega E. Full-mouth ultrasonic debridement versus quadrant scaling and root planing as an initial approach in the treatment of chronic periodontitis. J Clin Periodontol. 2005;32(8):851–9. 10.1111/j.1600-051X.2005.00776.x

4. Liu L, Guo S, Shi W, Liu Q, Huo F, Wu Y, et al. Bone marrow mesenchymal stem cell-derived small extracellular vesicles promote periodontal regeneration. Tissue Eng Part A. 2020;27(13–14):962–76. 10.1089/ten.TEA.2020.0141

5. Wang L, Yang Y, Xiong X, Yu T, Wang X, Meng W, et al. Oral lichen-planus-associated fibroblasts acquire myofibroblast characteristics and secrete pro-inflammatory cytokines in response to Porphyromonas gingivalis lipopolysaccharide stimulation. BMC Oral Health. 2018;18(1):197. 10.1186/s12903-018-0656-6

6. Bandow K, Maeda A, Kakimoto K, Kusuyama J, Shamoto M, Ohnishi T, et al. Molecular mechanisms of the inhibitory effect of lipopolysaccharide (LPS) on osteoblast differentiation. Biochem Biophys Res Commun. 2010;402(4):755–61. 10.1016/j.bbrc.2010.10.103

7. Zhou R, Chen F, Liu H, Zhu X, Wen X, Yu F, et al. Berberine ameliorates the LPS-induced imbalance of osteogenic and adipogenic differentiation in rat bone marrow-derived mesenchymal stem cells. Mol Med Rep. 2021;23(5):350. 10.3892/mmr.2021.11989

8. Wang X, Jiang M, He X, Zhang B, Peng W, Guo L. N-acetyl cysteine inhibits the lipopolysaccharide-induced inflammatory response in bone marrow mesenchymal stem cells by suppressing the TXNIP/NLRP3/IL-1β signaling pathway. Mol Med Rep. 2020;22(4):3299–306. 10.3892/mmr.2020.11433

9. Basri AM, Taha H, Ahmad N. A review on the pharmacological activities and phytochemicals of alpinia officinarum (Galangal) extracts derived from bioassay-guided fractionation and isolation. Pharmacogn Rev. 2017;11(21):43–56. 10.4103/phrev.phrev_55_16

10. Yang Z, Li X, Han W, Lu X, Jin S, Yang W, et al. Galangin suppresses human osteosarcoma cells: an exploration of its underlying mechanism. Oncol Rep. 2017;37(1):435–41. 10.3892/or.2016.5224

11. Song H-Y, Kim WS, Han JM, Seo HS, Lim S-T, Byun E-B. Galangin treatment during dendritic cell differentiation confers tolerogenic properties in response to lipopolysaccharide stimulation. J Nutr Biochem. 2021;87:108524. 10.1016/j.jnutbio.2020.108524

12. Choi M, Lee E-J, Park J-S, Kim S-N, Park E-M, Kim H-S. Anti-inflammatory mechanism of galangin in lipopolysaccha-ride-stimulated microglia: Critical role of PPAR-γ signaling pathway. Biochem Pharmacol. 2017;144 :120–31. 10.1016/j.bcp.2017.07.021

13. Kim J-H, Kang H-M, Yu S-B, Song J-M, Kim C-H, Kim B-J, et al. Cytoprotective effect of flavonoid-induced autophagy on bisphosphonate mediated cell death in osteoblast. J Cell Biochem. 2018;119(7):5571–80. 10.1002/jcb.26728

14. Huh J-E, In-Tae J, Junyoung C, Yong-Hyeon B, Jae-Dong L, Dong-Suk P, et al. The natural flavonoid galangin inhibits osteoclastic bone destruction and osteoclastogenesis by suppressing NF-κB in collagen-induced arthritis and bone marrow-derived macrophages. Eur J Pharmacol. 2012; 698(1–3):57–66. 10.1016/j.ejphar.2012.08.013

15. Liu C, Ma M, Zhang J, Gui S, Zhang X, Xue S. Galangin inhibits human osteosarcoma cells growth by inducing transforming growth factor-β1-dependent osteogenic differentiation. Biomed Pharmacother. 2017;89:1415–21. 10.1016/j.biopha.2017.03.030

16. Zhu J. Influences of traditional Chinese medicine intervention on the bone growth and metabolism of rats with simulated weightlessness. Asian Pac J Trop Med. 2013;6(3):224–7. 10.1016/S1995-7645(13)60028-0

17. Ki-Shuk Shim C-JL, Nam-Hui Yim, Min Jung Gu, Jin Yeul Ma. Alpinia officinarum stimulates osteoblast mineralization and inhibits osteoclast differentiation. Am J Chin Med. 2016;44(6): 1255–71. 10.1142/S0192415X16500701

18. Arvidson K, Abdallah BM, Applegate LA, Baldini N, Cenni E, Gomez-Barrena E, et al. Bone regeneration and stem cells. J Cell Mol Med. 2011;15(4):718–746. 10.1111/j.1582-4934.2010.01224.x

19. Lu L, Liu Y, Zhang X, Lin J. The therapeutic role of bone marrow stem cell local injection in rat experimental periodontitis. J Oral Rehabil. 2020;47(S1):73–82. 10.1111/joor.12843

20. Hienz SA, Paliwal S, Ivanovski S. Mechanisms of bone resorption in periodontitis. J Immunol Res. 2015;2015:615486. 10.1155/2015/615486

21. Huang R-L, Yuan Y, Zou G-M, Liu G, Tu J, Li Q. LPS-stimulated inflammatory environment inhibits BMP-2-induced osteoblastic differentiation through crosstalk between TLR4/MyD88/NF0ºB and BMP/Smad signaling. Stem Cells Dev. 2014;23(3):277–89. 10.1089/scd.2013.0345

22. Pogozhykh O, Pogozhykh D, Neehus A-L, Hoffmann A, Blasczyk R, Müller T. Molecular and cellular characteristics of human and non-human primate multipotent stromal cells from the amnion and bone marrow during long term culture. Stem Cell Res Ther. 2015;6(1):150. 10.1186/s13287-015-0146-6

23. Zhang L, Deng S. Effects of astragaloside IV on inflammation and immunity in rats with experimental periodontitis. Braz Oral Res. 2019;33:e032. 10.1590/1807-3107bor-2019.vol33.0032

24. Lee HN, Shin SA, Choo GS, Kim HJ, Park YS, Kim BS, et al. Anti-inflammatory effect of quercetin and galangin in LPS-stimulated RAW264.7 macrophages and DNCB-induced atopic dermatitis animal models.Int J Mol Med. 2018; 41(2):888–98. 10.3892/ijmm.2017.3296

25. Čebatariūnienė A, Kriaučiūnaitė K, Prunskaitė J, Tunaitis V, Pivoriūnas A. Extracellular vesicles suppress basal and lipopolysaccharide-induced NFκB activity in human periodontal ligament stem cells. Stem Cells Dev. 2019;28(15):1037–49. 10.1089/scd.2019.0021

26. Shu Y-S, Tao W, Miao Q-B, Lu S-C, Zhu Y-B. Galangin dampens mice lipopolysaccharide-induced acute lung injury. Inflammation. 2014;37(5):1661–8. 10.1007/s10753-014-9894-1

27. Sun K, Luo J, Guo J, Yao X, Jing X, Guo F. The PI3K/AKT/ mTOR signaling pathway in osteoarthritis: a narrative review. Osteoarthr Cartil. 2020;28(4):400–9. 10.1016/j.joca.2020.02.027

28. Hiraiwa M, Ozaki K, Yamada T, Iezaki T, Park G, Fukasawa K, et al. mTORC1 activation in osteoclasts prevents bone loss in a mouse model of osteoporosis. Front Pharmacol. 2019;10:684. 10.3389/fphar.2019.00684

29. Zhao S-J, Kong F-Q, Jie J, Li Q, Liu H, Xu A-D, et al. Macrophage MSR1 promotes BMSC osteogenic differentiation and M2-like polarization by activating PI3K/AKT/GSK3β/β-catenin pathway. Theranostics. 2020;10(1):17–35. 10.7150/thno.36930

30. Liu Y, Liang X, Zhang G, Kong L, Peng W, Zhang H. Galangin and pinocembrin from propolis ameliorate insulin resistance in HepG2 cells via regulating Akt/mTOR signaling. Evid Based Complement Alternat Med. 2018;2018:7971842. 10.1155/2018/7971842

31. Huang L, Lin M, Zhong X, Yang H, Deng M. Galangin decreases p-tau, Aβ42 and β-secretase levels, and suppresses autophagy in okadaic acid-induced PC12 cells via an Akt/GSK3β/ mTOR signaling-dependent mechanism. Mol Med Rep. 2019;19(3):1767–74. 10.3892/mmr.2019.9824