Abstract
Bone loss induced by microgravity is a substantial barrier to humans in long-term spaceflight. Recent studies have revealed that icariin (ICA) can attenuate osteoporosis in postmenopausal women and ovariectomized rats. However, whether ICA can protect against microgravity-induced bone loss remains unknown. In this study, the effects of ICA on a hindlimb suspension rodent model were investigated. Two-month-old female Wistar rats were hindlimb suspended and treated with ICA (25 mg·kg−1·d−1, i.g.) or a vehicle for 4 weeks (n = 6). The bone mass density of the hindlimbs was analyzed using dual-energy X-ray absorptiometry and micro-CT. mRNA expression of osteogenic genes in the tibia and the content of bone metabolism markers in serum were measured using qRT-PCR and ELISA, respectively. The bone mineral phase was analyzed using X-ray diffraction and atomic spectrometry. The results showed that ICA treatment significantly rescued the hindlimb suspension-induced reduction in bone mineral density, trabecular number and thickness, as well as the increases in trabecular separation and the structure model index. In addition, ICA treatment recovered the decreased bone-related gene expression, including alkaline phosphatase (ALP), bone glaprotein (BGP), and osteoprotegerin/receptor activator of the NF-κB ligand ratio (OPG/RANKL), in the tibia and the decreased bone resorption marker TRACP-5b levels in serum caused by simulated microgravity. Notably, ICA treatment restored the instability of bone biological apatite and the metabolic disorder of bone mineral elicited by simulated microgravity. These results demonstrate that ICA treatment plays osteoprotective roles in bone loss induced by simulated microgravity by inhibiting bone resorption and stabilizing bone biological apatite.
Similar content being viewed by others
Log in or create a free account to read this content
Gain free access to this article, as well as selected content from this journal and more on nature.com
or
References
Thirsk R, Kuipers A, Mukai C, Williams D. The space-flight environment: the International Space Station and beyond. CMAJ. 2009;180:1216–20.
Lang T, LeBlanc A, Evans H, Lu Y, Genant H, Yu A. Cortical and trabecular bone mineral loss from the spine and hip in long-duration spaceflight. J Bone Miner Res. 2004;19:1006–12.
Kondo H, Yumoto K, Alwood JS, Mojarrab R, Wang A, Almeida EA, et al. Oxidative stress and gamma radiation-induced cancellous bone loss with musculoskeletal disuse. J Appl Physiol. 2010;108:152–61.
Ya-Jing Y, Da-Chuan Y, Peng S. Effect of microgravity and a high magnetic field on hydroxyapatite deposition and implications for bone loss in space. Appl Surf Sci. 2010;256:7535–9.
Schwarzenberg M, Pippia P, Meloni MA, Cossu G, Cogoli-Greuter M, Cogoli A. Microgravity simulations with human lymphocytes in the free fall machine and in the random positioning machine. J Gravit Physiol. 1998;5:P23–6.
Hargens AR, Vico L. Long-duration bed rest as an analog to microgravity. J Appl Physiol. 2016;120:891–903.
Morey-Holton ER, Globus RK. Hindlimb unloading rodent model: technical aspects. J Appl Physiol. 2002;92:1367–77.
Convertino VA. Exercise as a countermeasure for physiological adaptation to prolonged spaceflight. Med Sci Sports Exerc. 1996;28:999–1014.
Cavanagh PR, Licata AA, Rice AJ. Exercise and pharmacological countermeasures for bone loss during long-duration space flight. Gravit Space Biol Bull. 2005;18:39–58.
Ettinger B, Genant HK, Cann CE. Postmenopausal bone loss is prevented by treatment with low-dosage estrogen with calcium. Ann Intern Med. 1987;106:40–5.
Reginster JY. The role of bisphosphonates in the prevention and treatment of osteoporosis. Clin Rheumatol. 1995;14 (Suppl 3):22–5.
Rizzoli R, Reginster JY, Boonen S, Breart G, Diez-Perez A, Felsenberg D, et al. Adverse reactions and drug-drug interactions in the management of women with postmenopausal osteoporosis. Calcif Tissue Int. 2011;89:91–104.
Rossouw JE, Anderson GL, Prentice RL, LaCroix AZ, Kooperberg C, Stefanick ML, et al. Risks and benefits of estrogen plus progestin in healthy postmenopausal women: principal results From the Women’s Health Initiative randomized controlled trial. JAMA. 2002;288:321–33.
Bolland MJ, Grey A, Avenell A, Gamble GD, Reid IR. Calcium supplements with or without vitamin D and risk of cardiovascular events: reanalysis of the Women’s Health Initiative limited access dataset and meta-analysis. BMJ. 2011;342:d2040.
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.
Zhang DC, Liu JL, Ding YB, Xia JG, Chen GY. Icariin potentiates the antitumor activity of gemcitabine in gallbladder cancer by suppressing NF-kappaB. Acta Pharmacol Sin. 2013;34:301–8.
Zhang G, Qin L, Shi Y. Epimedium-derived phytoestrogen flavonoids exert beneficial effect on preventing bone loss in late postmenopausal women: a 24-month randomized, double-blind and placebo-controlled trial. J Bone Miner Res. 2007;22:1072–9.
Chen G, Wang C, Wang J, Yin S, Gao H, Xiang LU, et al. Antiosteoporotic effect of icariin in ovariectomized rats is mediated via the Wnt/β-catenin pathway. Exp Ther Med. 2016;12:279–87.
Huang J, Yuan L, Wang X, Zhang TL, Wang K. Icaritin and its glycosides enhance osteoblastic, but suppress osteoclastic, differentiation and activity in vitro. Life Sci. 2007;81:832–40.
Zhao J, Ohba S, Shinkai M, Chung UI, Nagamune T. Icariin induces osteogenic differentiation in vitro in a BMP- and Runx2-dependent manner. Biochem Biophys Res Commun. 2008;369:444–8.
Chen KM, Ge BF, Liu XY, Ma PH, Lu MB, Bai MH, et al. Icariin inhibits the osteoclast formation induced by RANKL and macrophage-colony stimulating factor in mouse bone marrow culture. Pharmazie. 2007;62:388–91.
LeGeros RZ. Properties of osteoconductive biomaterials: calcium phosphates. Clin Orthop Relat Res. 2002;395:81–98.
Zioupos P, Currey JD, Hamer AJ. The role of collagen in the declining mechanical properties of aging human cortical bone. J Biomed Mater Res. 1999;45:108–16.
Bigi A, Foresti E, Gregorini R, Ripamonti A, Roveri N, Shah JS. The role of magnesium on the structure of biological apatites. Calcif Tissue Int. 1992;50:439–44.
Elliott JC, Anderson P, Gao XJ, Wong FS, Davis GR, Dowker SE. Application of scanning microradiography and x-ray microtomography to studies of bones and teeth. J X-ray Sci Technol. 1994;4:102–17.
Rey C. Calcium phosphate biomaterials and bone mineral. Differences in composition, structures and properties. Biomaterials. 1990;11:13–5.
Rey C, Combes C, Drouet C, Sfihi H, Barroug A. Physico-chemical properties of nanocrystalline apatites: Implications for biominerals and biomaterials. Mater Sci Eng: C. 2007;27:198–205.
Cheng K, Baofeng GE, Chen K, Zhen P, Zhou J, Xiaoni MA, et al. Oral medication of icariin enhances peak bone mineral density and bone quality in rats. Chin J Osteoporos. 2014;20:120–4.
Cheng K, Chen KM, Ge BF, Zhen P, Ma HP, Gao YH. Effect of icariin and genistein on bone protection. Chin Pharmacol Bull. 2014;30:1315–9.
Sun P, Liu Y, Deng X, Yu C, Dai N, Yuan X, et al. An inhibitor of cathepsin K, icariin suppresses cartilage and bone degradation in mice of collagen-induced arthritis. Phytomedicine. 2013;20:975–9.
Mkukuma LD, Skakle JM, Gibson IR, Imrie CT, Aspden RM, Hukins DW. Effect of the proportion of organic material in bone on thermal decomposition of bone mineral: an investigation of a variety of bones from different species using thermogravimetric analysis coupled to mass spectrometry, high-temperature X-ray diffraction, and Fourier transform infrared spectroscopy. Calcif Tissue Int. 2004;75:321–8.
Danilchenko SN, Koropov AV, Protsenko IY, Sulkio-Cleff B, Sukhodub LF. Thermal behavior of biogenic apatite crystals in bone: an X-ray diffraction study. Cryst Res Technol. 2006;41:268–75.
Danil’chenko SN, Kulik AN, Pavlenko PA, Kalinichenko TG, Bugai AN, Chemeris II, et al. Thermally activated diffusion of magnesium from bioapatite crystals. J Appl Spectrosc. 2006;73:437–43.
Li GW, Xu Z, Chang SX, Nian H, Wang XY, Qin LD. Icariin prevents ovariectomy-induced bone loss and lowers marrow adipogenesis. Menopause. 2014;21:1007–16.
Einhorn TA. Bone strength: the bottom line. Calcif Tissue Int. 1992;51:333–9.
Muller R, Hannan M, Smith SY, Bauss F. Intermittent ibandronate preserves bone quality and bone strength in the lumbar spine after 16 months of treatment in the ovariectomized cynomolgus monkey. J Bone Miner Res. 2004;19:1787–96.
Vasikaran S, Eastell R, Bruyere O, Foldes AJ, Garnero P, Griesmacher A, et al. Markers of bone turnover for the prediction of fracture risk and monitoring of osteoporosis treatment: a need for international reference standards. Osteoporos Int. 2011;22:391–420.
Garnero P. New developments in biological markers of bone metabolism in osteoporosis. Bone. 2014;66:46–55.
Halleen JM, Tiitinen SL, Ylipahkala H, Fagerlund KM, Vaananen HK. Tartrate-resistant acid phosphatase 5b (TRACP 5b) as a marker of bone resorption. Clin Lab. 2006;52:499–509.
North American Menopause S. The role of calcium in peri- and postmenopausal women: consensus opinion of The North American Menopause Society. Menopause. 2001;8:84–95.
Ohmori S, Kanda K, Kawano S, Kambe F, Seo H. Changes in calcium, PTH and 1,25(OH)2 vitamin D3 during tail-suspension in ovariectomized rats: effects of estrogen administration. Environ Med. 2000;44:75–8.
Ito T, Kurokouchi K, Ohmori S, Kanda K, Murata Y, Izumi R, et al. Changes in serum concentrations of calcium and its regulating hormones during tail suspension in rats. Environ Med. 1996;40:43–6.
Okyay E, Ertugrul C, Acar B, Sisman AR, Onvural B, Ozaksoy D. Comparative evaluation of serum levels of main minerals and postmenopausal osteoporosis. Maturitas. 2013;76:320–5.
Nian H, Ma MH, Nian SS, Xu LL. Antiosteoporotic activity of icariin in ovariectomized rats. Phytomedicine. 2009;16:320–6.
Danilchenko SN. The approach for determination of concentration and location of major impurities (Mg, Na, K) in biological apatite of mineralized tissues. J Nano-Electron Phys. 2013;5:171–93.
Rey C, Combes C, Drouet C, Lebugle A, Sfihi H, Barroug A. Nanocrystalline apatites in biological systems: characterisation, structure and properties. Materwiss Werksttech. 2007;38:996–1002.
Elliott JC. Structure and chemistry of the apatites and other calcium orthophosphates. Studies in inorganic chemistry. 1994;18:1–371
Acknowledgements
This work was supported by grants from the International Science & Technology Cooperation Program of China (ISTCP, No. 2015DFR30940), the Chinese Academy of Science (CAS) “Light of West China” Program (2015), Special Program from Chinese Academy of Science in Cooperation with Russia, Ukraine and the Republic of Belarus (2015, 2017), the Introduced Intelligence Project from the State Administration of Foreign Experts Affairs P.R. China (2016), and the Science and Technology Research Project of Gansu Province (Nos. 145RTSA012 and 17JR5RA307).
Author contributions
J-fW and J-pH designed the study; XF, J-pH, and W-gS performed the animal experiments; SD and AK contributed the XRD and mobile ionic species analysis; HL and MZ assisted in caring for the animals; XF, J-pH, and SD performed the data analyses; J-pH, XF, and J-fW wrote the manuscript.
Author information
Authors and Affiliations
Corresponding author
Ethics declarations
Competing interests
The authors declare no competing interests.
Rights and permissions
About this article
Cite this article
He, Jp., Feng, X., Wang, Jf. et al. Icariin prevents bone loss by inhibiting bone resorption and stabilizing bone biological apatite in a hindlimb suspension rodent model. Acta Pharmacol Sin 39, 1760–1767 (2018). https://doi.org/10.1038/s41401-018-0040-8
Received:
Accepted:
Published:
Version of record:
Issue date:
DOI: https://doi.org/10.1038/s41401-018-0040-8
Keywords
This article is cited by
-
Pharmacological inhibition of endoplasmic reticulum stress mitigates osteoporosis in a mouse model of hindlimb suspension
Scientific Reports (2024)
-
Enhancing osteogenic bioactivities of coaxial electrospinning nano-scaffolds through incorporating iron oxide nanoparticles and icaritin for bone regeneration
Nano Research (2024)
