The effects of Lycii Radicis Cortex on RANKL-induced osteoclast differentiation and activation in RAW 264.7 cells

Kim,Jae-Hyun; Kim,Eun-Young; Lee,Bina; Min,Ju-Hee; Song,Dea-Uk; Lim,Jeong-Min; Eom,Ji  Whan; Yeom,Mijung; Jung,Hyuk-Sang; Sohn,Youngjoo

doi:10.3892/ijmm.2016.2477

The effects of Lycii Radicis Cortex on RANKL-induced osteoclast differentiation and activation in RAW 264.7 cells

Authors:
- Jae-Hyun Kim
- Eun-Young Kim
- Bina Lee
- Ju-Hee Min
- Dea-Uk Song
- Jeong-Min Lim
- Ji Whan Eom
- Mijung Yeom
- Hyuk-Sang Jung
- Youngjoo Sohn
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Affiliations: Department of Anatomy, College of Korean Medicine, Kyung Hee University, Seoul 130-701, Republic of Korea, Acupuncture and Meridian Science Research Center, College of Korean Medicine, Kyung Hee University, Seoul 130-701, Republic of Korea
Published online on: February 1, 2016 https://doi.org/10.3892/ijmm.2016.2477
Pages: 649-658
Copyright: © Kim et al. This is an open access article distributed under the terms of Creative Commons Attribution License.

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Abstract

Post-menopausal osteoporosis is a serious age-related disease. After the menopause, estrogen deficiency is common, and excessive osteoclast activity causes osteoporosis. Osteoclasts are multinucleated cells generated from the differentiation of monocyte/macrophage precursor cells such as RAW 264.7 cells. The water extract of Lycii Radicis Cortex (LRC) is made from the dried root bark of Lycium chinense Mill. and is termed ‘Jigolpi’ in Korea. Its effects on osteoclastogenesis and post‑menopausal osteoporosis had not previously been tested. In the present study, the effect of LRC on receptor activator of nuclear factor-κB (NF-κB) ligand (RANKL)-induced osteoclast differentiation was demonstrated using a tartrate-resistant acid phosphatase (TRAP) assay and pit formation assay. Moreover, in order to analyze molecular mechanisms, we studied osteoclastogenesis-related markers such as nuclear factor of activated T-cells cytoplasmic 1 (NFATc1), c-Fos, receptor activator of NF-κB (RANK), TRAP, cathepsin K (CTK), matrix metallopeptidase-9 (MMP-9), calcitonin receptor (CTR) and carbonic anhydrase Ⅱ (CAII) using RT-qPCR and western blot analysis. Additionally, we also determined the effect of LRC on an ovariectomized (OVX) rat model. We noted that LRC inhibited RANKL-induced osteoclast differentiation via suppressing osteoclastogenesis-related markers. It also inhibited osteoporosis in the OVX rat model by decreasing loss of bone density and trabecular area. These results suggest that LRC exerts a positive effect on menopausal osteoporosis.

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1	Burge R, Dawson-Hughes B, Solomon DH, Wong JB, King A and Tosteson A: Incidence and economic burden of osteoporosis-related fractures in the United States, 2005–2025. J Bone Miner Res. 22:465–475. 2007. View Article : Google Scholar
2	Teitelbaum SL: Bone resorption by osteoclasts. Science. 289:1504–1508. 2000. View Article : Google Scholar : PubMed/NCBI
3	Rodan GA and Martin TJ: Therapeutic approaches to bone diseases. Science. 289:1508–1514. 2000. View Article : Google Scholar : PubMed/NCBI
4	Boyle WJ, Simonet WS and Lacey DL: Osteoclast differentiation and activation. Nature. 423:337–342. 2003. View Article : Google Scholar : PubMed/NCBI
5	Wensel TM, Iranikhah MM and Wilborn TW: Effects of denosumab on bone mineral density and bone turnover in postmenopausal women. Pharmacotherapy. 31:510–523. 2011. View Article : Google Scholar : PubMed/NCBI
6	de Villiers TJ: Bone health and osteoporosis in postmenopausal women. Best Pract Res Clin Obstet Gynaecol. 23:73–85. 2009. View Article : Google Scholar
7	Collin-Osdoby P and Osdoby P: RANKL-mediated osteoclast formation from murine RAW 264.7 cells. Methods Mol Biol. 816:187–202. 2012. View Article : Google Scholar
8	Suda T, Takahashi N, Udagawa N, Jimi E, Gillespie MT and Martin TJ: Modulation of osteoclast differentiation and function by the new members of the tumor necrosis factor receptor and ligand families. Endocr Rev. 20:345–357. 1999. View Article : Google Scholar : PubMed/NCBI
9	Grigoriadis AE, Wang ZQ, Cecchini MG, Hofstetter W, Felix R, Fleisch HA and Wagner EF: c-Fos: a key regulator of osteoclast-macrophage lineage determination and bone remodeling. Science. 266:443–448. 1994. View Article : Google Scholar : PubMed/NCBI
10	Zhao Q, Wang X, Liu Y, He A and Jia R: NFATc1: functions in osteoclasts. Int J Biochem Cell Biol. 42:576–579. 2010. View Article : Google Scholar
11	Choi HJ, Park YR, Nepal M, Choi BY, Cho NP, Choi SH, Heo SR, Kim HS, Yang MS and Soh Y: Inhibition of osteoclastogenic differentiation by Ikarisoside A in RAW 264.7 cells via JNK and NF-kappaB signaling pathways. Eur J Pharmacol. 636:28–35. 2010. View Article : Google Scholar : PubMed/NCBI
12	Fujisaki K, Tanabe N, Suzuki N, Kawato T, Takeichi O, Tsuzukibashi O, Makimura M, Ito K and Maeno M: Receptor activator of NF-kappaB ligand induces the expression of carbonic anhydrase II, cathepsin K, and matrix metalloproteinase-9 in osteoclast precursor RAW264.7 cells. Life Sci. 80:1311–1318. 2007. View Article : Google Scholar : PubMed/NCBI
13	Wimalawansa SJ: Prevention and treatment of osteoporosis: efficacy of combination of hormone replacement therapy with other antiresorptive agents. J Clin Densitom. 3:187–201. 2000. View Article : Google Scholar : PubMed/NCBI
14	Alexandersen P, Toussaint A, Christiansen C, Devogelaer JP, Roux C, Fechtenbaum J, Gennari C and Reginster JY; Ipriflavone Multicenter European Fracture S; Ipriflavone Multicenter European Fracture Study: Ipriflavone in the treatment of postmenopausal osteoporosis: a randomized controlled trial. JAMA. 285:1482–1488. 2001. View Article : Google Scholar : PubMed/NCBI
15	Herbal Pharmacology Compilation Committee: Herbal Pharmacology. Shinil books, Seoul. 715–717. 2010.
16	Herbology Editorial Committee of Korean Medicine schools: Boncho-hak [Herbology]. Younglimsa; Seoul: pp. 534–535. 2004
17	Song MY, Jung HW, Kang SY, Kim KH and Park YK: Anti-inflammatory effect of Lycii radicis in LPS-stimulated RAW 264.7 macrophages. Am J Chin Med. 42:891–904. 2014. View Article : Google Scholar : PubMed/NCBI
18	Cho SH, Park EJ, Kim EO and Choi SW: Study on the hypochlolesterolemic and antioxidative effects of tyramine derivatives from the root bark of Lycium chenese Miller. Nutr Res Pract. 5:412–420. 2011. View Article : Google Scholar : PubMed/NCBI
19	Kim SJ, Lee L, Kim JH, Lee TH and Shim I: Antidepressant-like effects of lycii radicis cortex and betaine in the forced swimming test in rats. Biomol Ther (Seoul). 21:79–83. 2013. View Article : Google Scholar
20	Jeong JC, Kim SJ, Kim YK, Kwon CH and Kim KH: Lycii cortex radicis extract inhibits glioma tumor growth in vitro and in vivo through downregulation of the Akt/ERK pathway. Oncol Rep. 27:1467–1474. 2012.PubMed/NCBI
21	Gao D, Li Q, Liu Z, Li Y, Liu Z, Fan Y, Li K, Han Z and Li J: Hypoglycemic effects and mechanisms of action of Cortex Lycii Radicis on alloxan-induced diabetic mice. Yakugaku Zasshi. 127:1715–1721. 2007. View Article : Google Scholar : PubMed/NCBI
22	Kalu DN: The ovariectomized rat model of postmenopausal bone loss. Bone Miner. 15:175–191. 1991. View Article : Google Scholar : PubMed/NCBI
23	Arjmandi BH, Alekel L, Hollis BW, Amin D, Stacewicz-Sapuntzakis M, Guo P and Kukreja SC: Dietary soybean protein prevents bone loss in an ovariectomized rat model of osteoporosis. J Nutr. 126:161–167. 1996.PubMed/NCBI
24	Hsu H, Lacey DL, Dunstan CR, Solovyev I, Colombero A, Timms E, Tan HL, Elliott G, Kelley MJ, Sarosi I, et al: Tumor necrosis factor receptor family member RANK mediates osteoclast differentiation and activation induced by osteoprotegerin ligand. Proc Natl Acad Sci USA. 96:3540–3545. 1999. View Article : Google Scholar : PubMed/NCBI
25	Tanaka H, Tanabe N, Shoji M, Suzuki N, Katono T, Sato S, Motohashi M and Maeno M: Nicotine and lipopolysaccharide stimulate the formation of osteoclast-like cells by increasing macrophage colony-stimulating factor and prostaglandin E2 production by osteoblasts. Life Sci. 78:1733–1740. 2006. View Article : Google Scholar
26	Tanabe N, Maeno M, Suzuki N, Fujisaki K, Tanaka H, Ogiso B and Ito K: IL-1 alpha stimulates the formation of osteoclast-like cells by increasing M-CSF and PGE₂ production and decreasing OPG production by osteoblasts. Life Sci. 77:615–626. 2005. View Article : Google Scholar : PubMed/NCBI
27	Kim SN, Kim MH, Min YK and Kim SH: Licochalcone A inhibits the formation and bone resorptive activity of osteoclasts. Cell Biol Int. 32:1064–1072. 2008. View Article : Google Scholar : PubMed/NCBI
28	Jun AY, Kim HJ, Park KK, Son KH, Lee DH, Woo MH, Kim YS, Lee SK and Chung WY: Extract of Magnoliae Flos inhibits ovariectomy-induced osteoporosis by blocking osteoclastogenesis and reducing osteoclast-mediated bone resorption. Fitoterapia. 83:1523–1531. 2012. View Article : Google Scholar : PubMed/NCBI
29	Kim K, Lee SH, Ha Kim J, Choi Y and Kim N: NFATc1 induces osteoclast fusion via up-regulation of Atp6v0d2 and the dendritic cell-specific transmembrane protein (DC-STAMP). Mol Endocrinol. 22:176–185. 2008. View Article : Google Scholar
30	Winslow MM, Pan M, Starbuck M, Gallo EM, Deng L, Karsenty G and Crabtree GR: Calcineurin/NFAT signaling in osteoblasts regulates bone mass. Dev Cell. 10:771–782. 2006. View Article : Google Scholar : PubMed/NCBI
31	Takahashi S, Goldring S, Katz M, Hilsenbeck S, Williams R and Roodman GD: Downregulation of calcitonin receptor mRNA expression by calcitonin during human osteoclast-like cell differentiation. J Clin Invest. 95:167–171. 1995. View Article : Google Scholar : PubMed/NCBI
32	Wang ZQ, Ovitt C, Grigoriadis AE, Möhle-Steinlein U, Rüther U and Wagner EF: Bone and haematopoietic defects in mice lacking c-fos. Nature. 360:741–745. 1992. View Article : Google Scholar : PubMed/NCBI
33	David JP, Rincon M, Neff L, Horne WC and Baron R: Carbonic anhydrase II is an AP-1 target gene in osteoclasts. J Cell Physiol. 188:89–97. 2001. View Article : Google Scholar : PubMed/NCBI
34	Sundaram K, Nishimura R, Senn J, Youssef RF, London SD and Reddy SV: RANK ligand signaling modulates the matrix metalloproteinase-9 gene expression during osteoclast differentiation. Exp Cell Res. 313:168–178. 2007. View Article : Google Scholar
35	Andersen TL, del Carmen Ovejero M, Kirkegaard T, Lenhard T, Foged NT and Delaissé JM: A scrutiny of matrix metalloproteinases in osteoclasts: evidence for heterogeneity and for the presence of MMPs synthesized by other cells. Bone. 35:1107–1119. 2004. View Article : Google Scholar : PubMed/NCBI
36	Versi E, Harvey MA, Cardozo L, Brincat M and Studd JW: Urogenital prolapse and atrophy at menopause: a prevalence study. Int Urogynecol J Pelvic Floor Dysfunct. 12:107–110. 2001. View Article : Google Scholar : PubMed/NCBI
37	Lim DW, Kim JG, Lee Y, Cha SH and Kim YT: Preventive effects of Eleutherococcus senticosus bark extract in OVX-induced osteoporosis in rats. Molecules. 18:7998–8008. 2013. View Article : Google Scholar : PubMed/NCBI
38	Lim DW and Kim YT: Dried root of Rehmannia glutinosa prevents bone loss in ovariectomized rats. Molecules. 18:5804–5813. 2013. View Article : Google Scholar : PubMed/NCBI
39	Hidaka S, Okamoto Y, Nakajima K, Suekawa M and Liu SY: Preventive effects of traditional Chinese (Kampo) medicines on experimental osteoporosis induced by ovariectomy in rats. Calcif Tissue Int. 61:239–246. 1997. View Article : Google Scholar : PubMed/NCBI
40	Dang ZC, van Bezooijen RL, Karperien M, Papapoulos SE and Löwik CW: Exposure of KS483 cells to estrogen enhances osteogenesis and inhibits adipogenesis. J Bone Miner Res. 17:394–405. 2002. View Article : Google Scholar : PubMed/NCBI
41	Wronski TJ, Lowry PL, Walsh CC and Ignaszewski LA: Skeletal alterations in ovariectomized rats. Calcif Tissue Int. 37:324–328. 1985. View Article : Google Scholar : PubMed/NCBI
42	Kalu DN, Liu CC, Salerno E, Hollis B, Echon R and Ray M: Skeletal response of ovariectomized rats to low and high doses of 17 beta-estradiol. Bone Miner. 14:175–187. 1991. View Article : Google Scholar : PubMed/NCBI
43	Hui SL, Slemenda CW and Johnston CC Jr: Baseline measurement of bone mass predicts fracture in white women. Ann Intern Med. 111:355–361. 1989. View Article : Google Scholar : PubMed/NCBI