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Journal of Orthopaedic Surgery and Research

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Open Access 01-12-2024 | Osteoporosis | Research article

Icariin promotes bone marrow mesenchymal stem cells osteogenic differentiation via the mTOR/autophagy pathway to improve ketogenic diet-associated osteoporosis

Authors: Wei Liu, Shouyu Xiang, Yingcong Wu, Dinghao Zhang, Chuhai Xie, Hailan Hu, Qi Liu

Published in: Journal of Orthopaedic Surgery and Research | Issue 1/2024

Abstract

Background

Icariin, a traditional Chinese medicine, has demonstrated anti-osteoporotic properties in ovariectomized mice. However, its effectiveness in preventing bone loss induced by ketogenic diet (KD), which mimics osteoporosis in human, remains unexplored. This study aims to investigate icariin’s impact on KD-induced bone loss in mice.

Methods

Thirty mice were divided into: sham, KD, and KD + icariin groups. Post a 12-week intervention, evaluation including bone microstructures, serum concentrations of tartrate-resistant acid phosphatase (TRAP) and bone-specific alkaline phosphatase (ALP), and femoral tissue expression levels of osteocalcin (OCN) and TRAP. The expression levels of mammalian target of rapamycin (mTOR), ALP, peroxisome proliferator-activated receptor gamma (PPAR-γ), phosphorylated mTOR (p-mTOR), and the autophagy adaptor protein (p62) were also analyzed. Alizarin granule deposition and cellular ALP levels were measured following the induction of bone marrow mesenchymal stem cells (BMSCs) into osteogenesis.

Results

The study found that KD significantly impaired BMSCs' osteogenic differentiation, leading to bone loss. Icariin notably increased bone mass, stimulated osteogenesis, and reduced cancellous bone loss. In the KD + icariin group, measures such as bone tissue density (TMD), bone volume fraction (BV/TV), trabecular number (Tb.N), and trabecular thickness (Tb.Th) were significantly higher than in the KD group. Additionally, bone trabecular separation (Tb.Sp) was markedly lower in the KD + icariin group. Moreover, icariin increased OCN and ALP levels while suppressing PPAR-γ, TRAP, p62, and p-mTOR. In cellular studies, icariin encouraged osteogenic development in BMSCs under KD conditions.

Conclusions

Icariin effectively counteracts bone thinning and improves bone microstructure. Its mechanism likely involves stimulating BMSCs osteogenic differentiation and inhibiting bone resorption, potentially through mTOR downregulation. These findings suggest icariin's potential as an alternative treatment for KD-induced bone loss.

Kushchayeva Y, Pestun I, Kushchayev S, Radzikhovska N, Lewiecki EM. Advancement in the treatment of osteoporosis and the effects on bone healing. J Clin Med. 2022;11(24):7477. https://doi.org/10.3390/jcm11247477.CrossRefPubMedPubMedCentral

Migliorini F, Giorgino R, Hildebrand F, Spiezia F, Peretti GM, Alessandri-Bonetti M, et al. Fragility fractures: risk factors and management in the elderly. Medicina (Kaunas). 2021;57(10):1119. https://doi.org/10.3390/medicina57101119.CrossRefPubMed

Lelovas PP, Xanthos TT, Thoma SE, Lyritis GP, Dontas IA. The laboratory rat as an animal model for osteoporosis research. Comp Med. 2008;58:424–30.PubMedPubMedCentral

Migliorini F, Colarossi G, Eschweiler J, Oliva F, Driessen A, Maffulli N. Antiresorptive treatments for corticosteroid-induced osteoporosis: a Bayesian network meta-analysis. Br Med Bull. 2022;143(1):46–56. https://doi.org/10.1093/bmb/ldac017.CrossRefPubMedPubMedCentral

Migliorini F, Maffulli N, Colarossi G, Eschweiler J, Tingart M, Betsch M. Effect of drugs on bone mineral density in postmenopausal osteoporosis: a Bayesian network meta-analysis. J Orthop Surg Res. 2021;16(1):533. https://doi.org/10.1186/s13018-021-02678-x.CrossRefPubMedPubMedCentral

Migliorini F, Maffulli N, Spiezia F, Peretti GM, Tingart M, Giorgino R. Potential of biomarkers during pharmacological therapy setting for postmenopausal osteoporosis: a systematic review. J Orthop Surg Res. 2021;16(1):351. https://doi.org/10.1186/s13018-021-02497-0.CrossRefPubMedPubMedCentral

Migliorini F, Maffulli N, Spiezia F, Tingart M, Maria PG, Riccardo G. Biomarkers as therapy monitoring for postmenopausal osteoporosis: a systematic review. J Orthop Surg Res. 2021;16(1):318. https://doi.org/10.1186/s13018-021-02474-7.CrossRefPubMedPubMedCentral

Conti V, Russomanno G, Corbi G, Toro G, Simeon V, Filippelli W, et al. A polymorphism at the translation start site of the vitamin D receptor gene is associated with the response to anti-osteoporotic therapy in postmenopausal women from southern Italy. Int J Mol Sci. 2015;16(3):5452–66. https://doi.org/10.3390/ijms16035452.CrossRefPubMedPubMedCentral

Migliorini F, Colarossi G, Baroncini A, Eschweiler J, Tingart M, Maffulli N. Pharmacological management of postmenopausal osteoporosis: a level I evidence based: expert opinion. Expert Rev Clin Pharmacol. 2021;14(1):105–19. https://doi.org/10.1080/17512433.2021.1851192.CrossRefPubMed

10.

Reid IR, Billington EO. Drug therapy for osteoporosis in older adults. Lancet. 2022;399(10329):1080–92. https://doi.org/10.1016/S0140-6736(21)02646-5.CrossRefPubMed

11.

An J, Yang H, Zhang Q, Liu C, Zhao J, Zhang L, et al. Natural products for treatment of osteoporosis: the effects and mechanisms on promoting osteoblast-mediated bone formation. Life Sci. 2016;147:46–58. https://doi.org/10.1016/j.lfs.2016.01.024.CrossRefPubMed

12.

Komori T. Animal models for osteoporosis. Eur J Pharmacol. 2015;759:287–94. https://doi.org/10.1016/j.ejphar.2015.03.028.ADSCrossRefPubMed

13.

Halade GV, Rahman MM, Williams PJ, Fernandes G. High fat diet-induced animal model of age-associated obesity and osteoporosis. J Nutr Biochem. 2010;21(12):1162–9. https://doi.org/10.1016/j.jnutbio.2009.10.002.CrossRefPubMedPubMedCentral

14.

Wu X, Huang Z, Wang X, Fu Z, Liu J, Huang Z, et al. Ketogenic diet compromises both cancellous and cortical bone mass in mice. Calcif Tissue Int. 2017;101(4):9412–21. https://doi.org/10.1007/s00223-017-0292-1.CrossRef

15.

Dowis K, Banga S. The potential health benefits of the ketogenic diet: a narrative review. Nutrients. 2021;13(5):1654. https://doi.org/10.3390/nu13051654.CrossRefPubMedPubMedCentral

16.

Xu X, Ding J, Wu X, Huang Z, Kong G, Liu Q, et al. Bone microstructure and metabolism changes under the combined intervention of ketogenic diet with intermittent fasting: an in vivo study of rats. Exp Anim. 2019;68(3):371–80. https://doi.org/10.1538/expanim.18-0084.CrossRefPubMedPubMedCentral

17.

Yin X, Zhou C, Li J, Liu R, Shi B, Yuan Q, et al. Autophagy in bone homeostasis and the onset of osteoporosis. Bone Res. 2019;7:28. https://doi.org/10.1038/s41413-019-0058-7.CrossRefPubMedPubMedCentral

18.

Chen X, Yang K, Jin X, Meng Z, Liu B, Yu H, et al. Bone autophagy: a potential way of exercise-mediated Meg3/P62/Runx2 pathway to regulate bone formation in T2DM mice. Diabetes Metab Syndr Obes. 2021;14:2753–64. https://doi.org/10.2147/DMSO.S299744.CrossRefPubMedPubMedCentral

19.

Li C, Li Q, Mei Q, Lu T. Pharmacological effects and pharmacokinetic properties of icariin, the major bioactive component in Herba Epimedii. Life Sci. 2015;126:57–68. https://doi.org/10.1016/j.lfs.2015.01.006.CrossRefPubMed

20.

Wang Z, Wang D, Yang D, Zhen W, Zhang J, Peng S. The effect of icariin on bone metabolism and its potential clinical application. Osteoporos Int. 2018;29(3):535–44. https://doi.org/10.1007/s00198-017-4255-1.CrossRefPubMed

21.

Cao H, Zhang Y, Qian W, Guo XP, Sun C, Zhang L, et al. Effect of icariin on fracture healing in an ovariectomized rat model of osteoporosis. Exp Ther Med. 2017;13(5):2399–404. https://doi.org/10.3892/etm.2017.4233.CrossRefPubMedPubMedCentral

22.

Liang X, Hou Z, Xie Y, Yan F, Li S, Zhu X, et al. Icariin promotes osteogenic differentiation of bone marrow stromal cells and prevents bone loss in OVX mice via activating autophagy. J Cell Biochem. 2019;120(8):13121–32. https://doi.org/10.1002/jcb.28585.CrossRefPubMed

23.

Tang Y, Li Y, Xin D, Chen L, Xiong Z, Yu X. Icariin alleviates osteoarthritis by regulating autophagy of chondrocytes by mediating PI3K/AKT/mTOR signaling. Bioengineered. 2021;12(1):2984–99. https://doi.org/10.1080/21655979.2021.1943602.CrossRefPubMedPubMedCentral

24.

Reeves PG. Components of the AIN-93 diets as improvements in the AIN-76A diet. J Nutr. 1997;127(5 Suppl):838S-841S. https://doi.org/10.1093/jn/127.5.838S.CrossRefPubMed

25.

Liu Q, Xu X, Yang Z, Liu Y, Wu X, Huang Z, et al. Metformin alleviates the bone loss induced by ketogenic diet: an in vivo study in mice. Calcif Tissue Int. 2019;104(1):59–69. https://doi.org/10.1007/s00223-018-0468-3.CrossRefPubMed

26.

Chappard D, Basle MF, Legrand E, Audran M. Trabecular bone microarchitecture: a review. Morphologie. 2008;92(299):162–70. https://doi.org/10.1016/j.morpho.2008.10.003.CrossRefPubMed

27.

Eastell R, Szulc P. Use of bone turnover markers in postmenopausal osteoporosis. Lancet Diabetes Endocrinol. 2017;5(11):908–23. https://doi.org/10.1016/S2213-8587(17)30184-5.CrossRefPubMed

28.

Noh JY, Yang Y, Jung H. Molecular mechanisms and emerging therapeutics for osteoporosis. Int J Mol Sci. 2020;21(20):7623. https://doi.org/10.3390/ijms21207623.CrossRefPubMedPubMedCentral

29.

Brown JP, Don-Wauchope A, Douville P, Albert C, Vasikaran SD. Current use of bone turnover markers in the management of osteoporosis. Clin Biochem. 2022;109–110:1–10. https://doi.org/10.1016/j.clinbiochem.2022.09.002.CrossRefPubMed

30.

Nuttall ME, Shah F, Singh V, Thomas-Porch C, Frazier T, Gimble JM. Adipocytes and the regulation of bone remodeling: a balancing act. Calcif Tissue Int. 2014;94(1):78–87. https://doi.org/10.1007/s00223-013-9807-6.CrossRefPubMed

31.

Szulc P. Bone turnover: biology and assessment tools. Best Pract Res Clin Endocrinol Metab. 2018;32(5):725–38. https://doi.org/10.1016/j.beem.2018.05.003.CrossRefPubMed

32.

Qadir A, Liang S, Wu Z, Chen Z, Hu L, Qian A. Senile osteoporosis: the involvement of differentiation and senescence of bone marrow stromal cells. Int J Mol Sci. 2020;21(1):349. https://doi.org/10.3390/ijms21010349.CrossRefPubMedPubMedCentral

33.

He C, Wang Z, Shi J. Pharmacological effects of icariin. Adv Pharmacol. 2020;87:179–203. https://doi.org/10.1016/bs.apha.2019.10.004.CrossRefPubMed

34.

Ming LG, Chen KM, Xian CJ. Functions and action mechanisms of flavonoids genistein and icariin in regulating bone remodeling. J Cell Physiol. 2013;228(3):513–21. https://doi.org/10.1002/jcp.24158.CrossRefPubMed

35.

Shen G, Ren H, Qiu T, Zhang Z, Zhao W, Yu X, et al. Mammalian target of rapamycin as a therapeutic target in osteoporosis. J Cell Physiol. 2018;233(5):3929–44. https://doi.org/10.1002/jcp.26161.CrossRefPubMed

36.

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. https://doi.org/10.1016/j.joca.2020.02.027.CrossRef

37.

Kim J, Kundu M, Viollet B, Guan KL. AMPK and mTOR regulate autophagy through direct phosphorylation of Ulk1. Nat Cell Biol. 2011;13(2):132–41. https://doi.org/10.1038/ncb2152.CrossRefPubMedPubMedCentral

Title: Icariin promotes bone marrow mesenchymal stem cells osteogenic differentiation via the mTOR/autophagy pathway to improve ketogenic diet-associated osteoporosis
Authors: Wei Liu
Shouyu Xiang
Yingcong Wu
Dinghao Zhang
Chuhai Xie
Hailan Hu
Qi Liu
Publication date: 01-12-2024
Publisher: BioMed Central
Keywords: Osteoporosis
Osteoporosis
Published in: Journal of Orthopaedic Surgery and Research / Issue 1/2024
Electronic ISSN: 1749-799X
DOI: https://doi.org/10.1186/s13018-024-04529-x

Keynote webinar | Spotlight on medication adherence

Springer Medicine

Icariin promotes bone marrow mesenchymal stem cells osteogenic differentiation via the mTOR/autophagy pathway to improve ketogenic diet-associated osteoporosis

Abstract

Background

Methods

Results

Conclusions

Keynote webinar | Spotlight on medication adherence

Springer Medicine

Abstract

Background

Methods

Results

Conclusions

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