Effects of a Hot-Water Extract of Allium hookeri Roots on Bone Formation in Human Osteoblast-Like MG-63 Cells In Vitro and in Rats In Vivo

Abstract

Allium hookeri is a wild herb found mainly in the Himalayas, growing at altitudes of 1400–4200 m. A. hookeri is widely consumed as a vegetable and herbal medicine in Asia, but its effects on bone health have not been reported previously. This study investigated the effects of a hot-water extract of A. hookeri roots on bone formation. The hot-water extract significantly increased the proliferation of in vitro human osteoblast-like MG-63 cells and the stimulatory effects on osteoblast differentiation were noticeably greater for the hot-water extract than for daidzein (a positive control), as reflected by alkaline phosphatase activity, collagen content, and mineral deposition. Expression of the bone-remodeling marker osteocalcin production and bone microstructural parameters were significantly improved in Sprague-Dawley rats in vivo after oral treatment with the hot-water extract compared with their control (saline-administered) counterparts. The chemical compounds of the hot-water extract were characterized by liquid chromatography-mass spectrometry, and alliin, sinapic acid, and ferulic acid, which exert beneficial effects on bone health, were identified. These findings indicate that A. hookeri can be used as a natural resource for increasing bone formation. This is the first report of the anabolic effects of A. hookeri extracts on bone formation in vitro and in vivo.

Key words

Allium hookeri - Liliaceae - bone formation - human osteoblast-like cell - osteocalcin - bone microstructure - chemical compound

• References

• 1 Hooker JD. The Flora of British India, Vol. VI. Dehradun: Bishen Singh Mahendrapal Singh; 1978
• 2 Ayam VS. Allium hookeri, Thw. Enum. A lesser known terrestrial perennial herb used as food and its ethnobotanical relevance in Manipur. Afr J Food Agric Nutr Dev 2011; 11: 5389-5412
• 3 Kala CP. Ethnomedicinal botany of the Apatani in the Eastern Himalayan region of India. J Ethnobiol Ethnomed 2005; 1: 11
• 4 Singh HB, Singh RS, Sandhu JS. Herbal Medicine of Manipur: A colour Encyclopaedia. Delhi: Daya Publishing House; 2003
• 5 Bae GC, Bae DY. The anti-inflammatory effects of ethanol extract of Allium hookeri cultivated in South Korea. Korea J Herbol 2012; 27: 55-61
• 6 Singh KB, Singh NM. Antioxidant and free radical scavenging potential of Allium hookeri Thwaites roots extract studied using in vitro models. J Adv Biol 2014; 4: 276-285
• 7 Won JY, Yoo YC, Kang EJ, Yang H, Kim GH, Seong BJ, Kim SI, Han SH, Lee SS, Lee KS. Chemical components, DPPH radical scavenging activity and inhibitory effects on nitric oxide production in Allium hookeri cultivated under open field and greenhouse conditions. J Korean Soc Food Sci Nutr 2013; 42: 1351-1356
• 8 Jeong J, Park H, Hyun H, Kim J, Kim H, Oh HI, Hwang HS, Kim DK, Kim HH. Effects of glucosinolates from turnip (Brassica rapa L.) root on bone formation by human osteoblast-like MG-63 cells and in normal young rats. Phytother Res 2015; 29: 902-909
• 9 Jia TL, Wang HZ, Xie LP, Wang XY, Zhang RQ. Daidzein enhances osteoblast growth that may be mediated by increased bone morphogenetic protein (BMP) production. Biochem Pharmacol 2003; 65: 709-715
• 10 Sugimoto E, Yamaguchi M. Stimulatory effect of daidzein in osteoblastic MC3T3-E1 cells. Biochem Pharmacol 2000; 59: 471-475
• 11 Huang W, Yang S, Shao J, Li YP. Signaling and transcriptional regulation in osteoblast commitment and differentiation. Front Biosci 2007; 12: 3068-3092
• 12 Zur Nieden NI, Kempka G, Ahr HJ. In vitro differentiation of embryonic stem cells into mineralized osteoblasts. Differentiation 2003; 71: 18-27
• 13 Declercq HA, Verbeeck RM, De Ridder LI, Schacht EH, Cornelissen MJ. Calcification as an indicator of osteoinductive capacity of biomaterials in osteoblastic cell cultures. Biomaterials 2005; 26: 4964-4974
• 14 Reagan-Shaw S, Nihal M, Ahmad N. Dose translation from animal to human studies revisited. FASEB J 2008; 22: 659-661
• 15 Delmas PD, Eastell R, Garnero P, Seibel MJ, Stepan J. The use of biochemical markers of bone turnover in osteoporosis. Osteoporos Int 2000; 11: S2-S17
• 16 Bouxsein ML, Boyd SK, Christiansen BA, Guldberg RE, Jepsen KJ, Müller R. Guidelines for assessment of bone microstructure in rodents using micro-computed tomography. J Bone Miner Res 2010; 25: 1468-1486
• 17 Sengupta P. The laboratory rat: relating its age with humanʼs. Int J Prev Med 2013; 4: 624-630
• 18 Fares JE, Choucair M, Nabulsi M, Salamoun M, Shahine CH, Fuleihana GEH. Effect of gender, puberty, and vitamin D status on biochemical markers of bone remodedeling. Bone 2003; 33: 242-247
• 19 Joseph EE, Gibbons M, Molina S. The effect of alliin on oxidative stress-induced bone-like cells (UMR 106-01 BSP). FASEB J 2013; 27: 744.6
• 20 Mahalakshmi G, Nirmala T, Samundeswari R, Sivassoupramanien D, Kavimani S. A review on medicinal plants for the management of osteoporosis. Biolife J 2014; 2: 1191-1198
• 21 Sassa S, Kikuchi T, Shinoda H, Suzuki S, Kudo H, Sakamoto S. Preventive effect of ferulic acid on bone loss in ovariectomized rats. In Vivo 2003; 17: 277-280
• 22 Franceschi RT, Iyer BS, Cui Y. Effects of ascorbic acid on collagen matrix formation and osteoblast differentiation in murine MC3T3-E1 cells. J Bone Miner Res 1994; 9: 843-854
• 23 Banu J, Varela E, Fernandes G. Alternative therapies for the prevention and treatment of osteoporosis. Nutr Rev 2012; 70: 22-40
• 24 Raggatt LJ, Partridge NC. Cellular and molecular mechanisms of bone remodeling. J Biol Chem 2010; 285: 25103-25108
• 25 Hinoi E, Fujimori S, Wang L, Hojo H, Uno K, Yoneda Y. Nrf2 negatively regulates osteoblast differentiation via interfering with Runx2-dependent transcriptional activation. J Biol Chem 2006; 281: 18015-18024
• 26 Wauquier F, Leotoing L, Coxam V, Guicheux J, Wittrant Y. Oxidative stress in bone remodelling and disease. Trends Mol Med 2009; 15: 468-477
• 27 Manolagas SC, Almeida M. Gone with the Wnts: beta-catenin, T-cell factor, forkhead box O, and oxidative stress in age-dependent diseases of bone, lipid and glucose metabolism. Mol Endocrinol 2007; 21: 2605-2614
• 28 Tominaga H, Ishiyama M, Ohseto F, Sasamoto K, Hamamoto T, Suzuki K, Watanabe M. A water-soluble tetrazolium salt useful for colorimetric cell viability assay. Anal Commun 1999; 36: 47-50
• 29 Abe T, Abe Y, Aida Y, Hara Y, Maeda K. Extracellular matrix regulates induction of alkaline phosphatase expression by ascorbic acid in human fibroblasts. J Cell Physiol 2001; 189: 144-151
• 30 Tullberg-Reinert H, Jundt G. In situ measurement of collagen synthesis by human bone cells with a Sirius Red-based colorimetric microassay: effects of transforming growth factor β2 and ascorbic acid 2-phosphate. Histochem Cell Biol 1999; 112: 271-276
• 31 Gregory CA, Gunn WG, Peister A, Prockop DJ. An Alizarin red-based assay of mineralization by adherent cells in culture: comparison with cetylpyridinium chloride extraction. Anal Biochem 2004; 329: 77-84
• 32 Rabenhorst A, Christopeit B, Leja S, Gerbaulet A, Kleiner S, Förster A, Raap U, Wickenhauser C, Hartmann K. Serum levels of bone cytokines are increased in indolent systemic mastocytosis associated with osteopenia or osteoporosis. J Allergy Clin Immunol 2013; 132: 1234-1237
• 33 Reinehr T, de Sousa G, Alexy U, Kersting M, Andler W. Vitamin D status and parathyroid hormone in obese children before and after weight loss. Eur J Endocrinol 2007; 157: 225-232
• 34 McCabe KM, Booth SL, Fu X, Shobeiri N, Pang JJ, Adams MA, Holden RM. Dietary vitamin K and therapeutic warfarin alter the susceptibility to vascular calcification in experimental chronic kidney disease. Kidney Int 2013; 83: 835-844