The antiosteoporosis effect of icariin in ovariectomized rats: a systematic review and meta-analysis

Yu Liu; Haojiang Zuo; Xuewei Liu; Jingyuan Xiong; Xiaofang Pei

doi:10.14715/cmb/2017.63.11.22

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Vol. 63 No. 11: Issue 11

Issue Published : November 30, 2017

The undersigned hereby assign all rights, included but not limited to copyright, for this manuscript to CMB Association upon its submission for consideration to publication on Cellular and Molecular Biology. The rights assigned include, but are not limited to, the sole and exclusive rights to license, sell, subsequently assign, derive, distribute, display and reproduce this manuscript, in whole or in part, in any format, electronic or otherwise, including those in existence at the time this agreement was signed. The authors hereby warrant that they have not granted or assigned, and shall not grant or assign, the aforementioned rights to any other person, firm, organization, or other entity. All rights are automatically restored to authors if this manuscript is not accepted for publication.

The antiosteoporosis effect of icariin in ovariectomized rats: a systematic review and meta-analysis

https://doi.org/10.14715/cmb/2017.63.11.22

Yu Liu

Department of Public Health Laboratory Sciences, West China School of Public Health, Sichuan University, Chengdu, 610041 China

Haojiang Zuo

Department of Public Health Laboratory Sciences, West China School of Public Health, Sichuan University, Chengdu, 610041 China

Xuewei Liu

Department of Public Health Laboratory Sciences, West China School of Public Health, Sichuan University, Chengdu, 610041 China

Jingyuan Xiong

Research Center for Public Health and Preventive Medicine, West China School of Public Health, Sichuan University, Chengdu, 610041 China

Xiaofang Pei

Research Center for Public Health and Preventive Medicine, West China School of Public Health, Sichuan University, Chengdu, 610041 China

Corresponding Author(s) : Jingyuan Xiong

jzx0004@tigermail.auburn.edu

Cellular and Molecular Biology, Vol. 63 No. 11: Issue 11
Article Published : November 30, 2017

Abstract

Herba Epimedii (Yinyanghuo or Horny Goat Weed) is a traditional Chinese medicine widely used in treating osteoporosis. As the major active component, icariin is intensively investigated in the prevention and treatment of osteoporosis in ovariectomized rats. However, misleading conclusions can be generated in animal studies with various experimental designs. Therefore, a systematic review and meta-analysis was performed to evaluate the efficacy of icariin against osteoporosis in ovariectomized rats. PubMed, EMBASE and Chinese National Knowledge Infrastructure databases were searched to identify studies of icariin in ovariectomized rats. Two independent authors selected and reviewed the publications. Data were pooled using a DerSimonian and Laird random-effects model. The results demonstrated that ovariectomized rats treated with icariin had significantly higher bone mineral density (femur and lumbar spine) and lower bone turnover markers (serum alkaline phosphatase and osteocalcin) compared with the ovariectomized control group. For bone histomorphometric parameters, the percentages of trabecular area and trabecular thickness were significantly higher while the trabecular separation was significantly lower in the ovariectomized rats treated with icariin. Based on these results, the presnet study suggested that icariin might possess substantial antiosteoporosis effect in ovariectomized rats. Safety studies and large randomized clinical trials are needed to further support possible clinical applications of icariin in postmenopausal women with osteoporosis.

Keywords

Icariin Osteoporosis Ovariectomized rats Systematic review Meta-analysis.

Liu, Y., Zuo, H., Liu, X., Xiong, J., & Pei, X. (2017). The antiosteoporosis effect of icariin in ovariectomized rats: a systematic review and meta-analysis. Cellular and Molecular Biology, 63(11), 124–131. https://doi.org/10.14715/cmb/2017.63.11.22

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References

Raisz LG. Pathogenesis of osteoporosis: concepts, conflicts, and prospects. J Clin Invest 2005; 115: 3318-3325.
NIH Consensus Development Panel on Osteoporosis Prevention, Diagnosis, and Therapy. Osteoporosis prevention, diagnosis, and therapy. JAMA 2001; 285: 785-795.
Hansen MA, Overgaard K, Riis BJ, Christiansen C. Potential risk factors for development of postmenopausal osteoporosis--examined over a 12-year period. Osteoporos Int 1991; 1: 95-102.
Lindsay R, Hart DM, Aitken JM, MacDonald EB, Anderson JB, Clarke AC. Long-term prevention of postmenopausal osteoporosis by oestrogen. Evidence for an increased bone mass after delayed onset of oestrogen treatment. Lancet 1976; 1: 1038-1041.
Fitzpatrick LA. Estrogen therapy for postmenopausal osteoporosis. Arq Bras Endocrinol Metabol 2006; 50: 705-719.
Lindsay R, Hart DM, Clark DM. The minimum effective dose of estrogen for prevention of postmenopausal bone loss. Obstet Gynecol 1984; 63: 759-763.
Blank RD, Bockman RS. A review of clinical trials of therapies for osteoporosis using fracture as an end point. J Clin Densitom 1999; 2: 435-452.
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: 321-333.
Beral V. Breast cancer and hormone-replacement therapy in the Million Women Study. Lancet 2003; 362: 419-427.
Gambacciani M, Ciaponi M, Genazzani AR. The HRT misuse and osteoporosis epidemic: a possible future scenario. Climacteric 2007; 10: 273-275.
Papaioannou A, Kennedy CC, Dolovich L, Lau E, Adachi JD. Patient adherence to osteoporosis medications: problems, consequences and management strategies. Drugs Aging 2007; 24: 37-55.
Watts NB, Diab DL. Long-term use of bisphosphonates in osteoporosis. J Clin Endocrinol Metab 2010; 95: 1555-1565.
O'Regan RM, Gradishar WJ. Selective estrogen-receptor modulators in 2001. Oncology (Williston Park) 2001; 15: 1177-1185, 89-90; discussion 90-94.
Body JJ. Calcitonin for the long-term prevention and treatment of postmenopausal osteoporosis. Bone 2002; 30: 75-79.
Watts NB. Bisphosphonate treatment of osteoporosis. Clin Geriatr Med 2003; 19: 395-414.
Zhai YK, Guo X, Pan YL, Niu YB, Li CR, Wu XL, et al. A systematic review of the efficacy and pharmacological profile of Herba Epimedii in osteoporosis therapy. Pharmazie 2013; 68: 713-722.
Feng D, Gao ZY. Determination of the content of icariin in Liuweixianshenzhi tablets with HPLC. Chin Pharm 2001; 12: 42-43.
Gao G, Liao M, Feng Y. Determination of flavonoids and quality evaluation of Chinese traditional drug "puhuang". Yao Xue Xue Bao 1998; 33: 300-303.
Pound P, Roberts I. The need for systematic reviews of animal studies. Chin J Evid Based Med 2005; 5: 3-5.
Macleod MR, O'Collins T, Howells DW, Donnan GA. Pooling of animal experimental data reveals influence of study design and publication bias. Stroke 2004; 35: 1203-1208.
Kilkenny C, Browne W, Cuthill IC, Emerson M, Altman DG. Animal research: reporting in vivo experiments: the ARRIVE guidelines. Br J Pharmacol 2010; 160: 1577-1579.
Higgins JP, Thompson SG. Quantifying heterogeneity in a meta-analysis. Stat Med 2002; 21: 1539-1558.
Zintzaras E, Ioannidis JP. Heterogeneity testing in meta-analysis of genome searches. Genet Epidemiol 2005; 28: 123-137.
Jove M, Pamplona R, Prat J, Arola L, Portero-Otin M. Atherosclerosis prevention by nutritional factors: a meta-analysis in small animal models. Nutr Metab Cardiovasc Dis 2013; 23: 84-93.
Wen J, Li YP. The selection of a summary statistic for use in meta-analysis. Chin J Evid Based Med 2007; 7: 606-613.
Nian H, Ma MH, Nian SS, Xu LL. Antiosteoporotic activity of icariin in ovariectomized rats. Phytomedicine 2009; 16: 320-326.
Qin L, Han T, Zhang Q, Cao D, Nian H, Rahman K, et al. Antiosteoporotic chemical constituents from Er-Xian Decoction, a traditional Chinese herbal formula. J Ethnopharmacol 2008; 118: 271-279.
Yang L, Yu Z, Qu H, Li M. Comparative effects of hispidulin, genistein, and icariin with estrogen on bone tissue in ovariectomized rats. Cell Biochem Biophys 2014; 70: 485-490.
Liu M, Zhong C, He RX, Chen LF. Icariin associated with exercise therapy is an effective treatment for postmenopausal osteoporosis. Chin Med J 2012; 125: 1784-1789.
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: 1007-1016.
Bian Q, Liu SF, HUang JH, Yang Z, Ning Y, Zhao YJ, Wang YJ, Shen ZY. Regulation and control of three active components of TCM effective in reinforcing kidney on ovariectomy-induced osteoporosis rats's bone marrow stem cells revealed by gene microarray. Chin J TCM 2011; 26: 889-893.
Qian GF, Yang H, Zhang XZ. Effect of icariin on the expression of Cbfa1in the bone of ovariectomized rats. Chin J Osteoporosis & Bone Miner Res 2009; 2: 104-109.
Wu MS, Zhao SZ, Ren LZ, Wang R, Bai X. Effects of icariin on expression of the estrogen receptor beta mRNA of bone and different hypothalamic nuclei in ovariectomy rats. Chin Pharm Bulletin 2011; 27: 29-33.
Bao JR, Yang JW, Li SF, Zhao W, Zhang QL, Yan YQ. Effects of icarrin on ovariectomized rats. J Hygene Res 2005; 34: 191-193.
Song Y, Li SH, He ZJ, Zhang YW, Zhou MW. Effect of icariin on osteoporosis in ovariectomized female rats. J Chin PLA Postgard Med Sch 2012; 33: 400-403.
Bai J. Effects of icariin on ovariectomized osteoporotic rats [Dissertation]. Lanzhou (China): Lanzhou University of Technology 2010.
Cheng K. Comparative research of icariin and genistein of pharmacological activity in antiosteoporosis [Dissertation]. Lanzhou (China): Lanzhou University of Technology 2014.
Liu SJ, Mo XM, Li JP, Yang J, Zhang XJ. Effecs of icariin and possible mechanisms in ovariectomized rats. Chin J Basic Med in TCM 2012; 3: 271-275.
Xue L, Wang Y, Jiang Y, Han T, Nie Y, Zhao L, et al. Comparative effects of er-xian decoction, epimedium herbs, and icariin with estrogen on bone and reproductive tissue in ovariectomized rats. Evid Based Complement Alternat Med 2012; 2012: 241416.
Aswar UM, Mohan V, Bodhankar SL. Antiosteoporotic activity of phytoestrogen-rich fraction separated from ethanol extract of aerial parts of Cissus quadrangularis in ovariectomized rats. Indian J Pharmacol 2012; 44: 345-350.
Kalu DN. The ovariectomized rat model of postmenopausal bone loss. Bone Miner 1991; 15: 175-191.
Manolagas SC, Jilka RL. Bone marrow, cytokines, and bone remodeling. Emerging insights into the pathophysiology of osteoporosis. N Engl J Med 1995; 332: 305-311.
Lerner UH. Bone remodeling in post-menopausal osteoporosis. J Dent Res 2006; 85: 584-595.
Mukaiyama K, Kamimura M, Uchiyama S, Ikegami S, Nakamura Y, Kato H. Elevation of serum alkaline phosphatase (ALP) level in postmenopausal women is caused by high bone turnover. Aging Clin Exp Res 2015; 27: 413-418.
Yogesh HS, Chandrashekhar VM, Katti HR, Ganapaty S, Raghavendra HL, Gowda GK, et al. Anti-osteoporotic activity of aqueous-methanol extract of Berberis aristata in ovariectomized rats. J Ethnopharmacol 2011; 134: 334-338.
Hertrampf T, Schleipen B, Offermanns C, Velders M, Laudenbach U, Diel P. Comparison of the bone protective effects of an isoflavone-rich diet with dietary and subcutaneous administrations of genistein in ovariectomized rats. Toxicol Lett 2009; 184: 198-203.
Qian WQ, Yin H, Sun HT. Influence of different concentrationsof icariin on osteoblast metabolism of rats. Chin Med Herald 2011; 8: 23-25.
Guo HL, Zhao YF, Wang X, Xu Y, Zhan HS. Experimental study on the mechanism of icariin improving human osteoblasts proliferation and the expression of OPG protein. Chin J Orthop Trauma 2011; 24: 585-588.
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: 832-840.
Cao H, Ke Y, Zhang Y, Zhang CJ, Qian W, Zhang GL. Icariin stimulates MC3T3-E1 cell proliferation and differentiation through up-regulation of bone morphogenetic protein-2. Int J Mol Med 2012; 29: 435-439.
Zhao J, Ohba S, Komiyama Y, Shinkai M, Chung UI, Nagamune T. Icariin: a potential osteoinductive compound for bone tissue engineering. Tissue engineering Part A 2010; 16: 233-243.
Mok SK, Chen WF, Lai WP, Leung PC, Wang XL, Yao XS, et al. Icariin protects against bone loss induced by oestrogen deficiency and activates oestrogen receptor-dependent osteoblastic functions in UMR 106 cells. Br J Pharmacol 2010; 159: 939-949.
Zheng D, Peng S, Yang SH, Shao ZW, Yang C, Feng Y, et al. The beneficial effect of Icariin on bone is diminished in osteoprotegerin-deficient mice. Bone 2012; 51: 85-92.
Hsieh TP, Sheu SY, Sun JS, Chen MH, Liu MH. Icariin isolated from Epimedium pubescens regulates osteoblasts anabolism through BMP-2, SMAD4, and Cbfa1 expression. Phytomedicine 2010; 17: 414-423.
Liang W, Lin M, Li X, Li C, Gao B, Gan H, et al. Icariin promotes bone formation via the BMP-2/Smad4 signal transduction pathway in the hFOB 1.19 human osteoblastic cell line. Int J Mol Med 2012; 30: 889-895.
Bagi C, van der Meulen M, Brommage R, Rosen D, Sommer A. The effect of systemically administered rhIGF-I/IGFBP-3 complex on cortical bone strength and structure in ovariectomized rats. Bone 1995; 16: 559-565.
Jee WS, Yao W. Overview: animal models of osteopenia and osteoporosis. J Musculoskelet Neuronal Interact 2001; 1: 193-207.
Pound P, Ebrahim S, Sandercock P, Bracken MB, Roberts I. Where is the evidence that animal research benefits humans? BMJ 2004; 328: 514-517.
van der Worp HB, Howells DW, Sena ES, Porritt MJ, Rewell S, O'Collins V, et al. Can animal models of disease reliably inform human studies? PLoS Med 2010; 7: e1000245.
Ma HP, Ming LG, Ge BF, Zhai YK, Song P, Xian CJ, et al. Icariin is more potent than genistein in promoting osteoblast differentiation and mineralization in vitro. J Cell Biochem 2011; 112: 916-923.
Bebarta V, Luyten D, Heard K. Emergency medicine animal research: does use of randomization and blinding affect the results? Acad Emerg Med 2003; 10: 684-687.
Sterne JA, Sutton AJ, Ioannidis JP, Terrin N, Jones DR, Lau J, et al. Recommendations for examining and interpreting funnel plot asymmetry in meta-analyses of randomised controlled trials. BMJ 2011; 343: d4002.
Macleod MR, O'Collins T, Horky LL, Howells DW, Donnan GA. Systematic review and metaanalysis of the efficacy of FK506 in experimental stroke. J Cereb Blood Flow Metab 2005; 25: 713-721.

References

Raisz LG. Pathogenesis of osteoporosis: concepts, conflicts, and prospects. J Clin Invest 2005; 115: 3318-3325.

NIH Consensus Development Panel on Osteoporosis Prevention, Diagnosis, and Therapy. Osteoporosis prevention, diagnosis, and therapy. JAMA 2001; 285: 785-795.

Hansen MA, Overgaard K, Riis BJ, Christiansen C. Potential risk factors for development of postmenopausal osteoporosis--examined over a 12-year period. Osteoporos Int 1991; 1: 95-102.

Lindsay R, Hart DM, Aitken JM, MacDonald EB, Anderson JB, Clarke AC. Long-term prevention of postmenopausal osteoporosis by oestrogen. Evidence for an increased bone mass after delayed onset of oestrogen treatment. Lancet 1976; 1: 1038-1041.

Fitzpatrick LA. Estrogen therapy for postmenopausal osteoporosis. Arq Bras Endocrinol Metabol 2006; 50: 705-719.

Lindsay R, Hart DM, Clark DM. The minimum effective dose of estrogen for prevention of postmenopausal bone loss. Obstet Gynecol 1984; 63: 759-763.

Blank RD, Bockman RS. A review of clinical trials of therapies for osteoporosis using fracture as an end point. J Clin Densitom 1999; 2: 435-452.

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: 321-333.

Beral V. Breast cancer and hormone-replacement therapy in the Million Women Study. Lancet 2003; 362: 419-427.

Gambacciani M, Ciaponi M, Genazzani AR. The HRT misuse and osteoporosis epidemic: a possible future scenario. Climacteric 2007; 10: 273-275.

Papaioannou A, Kennedy CC, Dolovich L, Lau E, Adachi JD. Patient adherence to osteoporosis medications: problems, consequences and management strategies. Drugs Aging 2007; 24: 37-55.

Watts NB, Diab DL. Long-term use of bisphosphonates in osteoporosis. J Clin Endocrinol Metab 2010; 95: 1555-1565.

O'Regan RM, Gradishar WJ. Selective estrogen-receptor modulators in 2001. Oncology (Williston Park) 2001; 15: 1177-1185, 89-90; discussion 90-94.

Body JJ. Calcitonin for the long-term prevention and treatment of postmenopausal osteoporosis. Bone 2002; 30: 75-79.

Watts NB. Bisphosphonate treatment of osteoporosis. Clin Geriatr Med 2003; 19: 395-414.

Zhai YK, Guo X, Pan YL, Niu YB, Li CR, Wu XL, et al. A systematic review of the efficacy and pharmacological profile of Herba Epimedii in osteoporosis therapy. Pharmazie 2013; 68: 713-722.

Feng D, Gao ZY. Determination of the content of icariin in Liuweixianshenzhi tablets with HPLC. Chin Pharm 2001; 12: 42-43.

Gao G, Liao M, Feng Y. Determination of flavonoids and quality evaluation of Chinese traditional drug "puhuang". Yao Xue Xue Bao 1998; 33: 300-303.

Pound P, Roberts I. The need for systematic reviews of animal studies. Chin J Evid Based Med 2005; 5: 3-5.

Macleod MR, O'Collins T, Howells DW, Donnan GA. Pooling of animal experimental data reveals influence of study design and publication bias. Stroke 2004; 35: 1203-1208.

Kilkenny C, Browne W, Cuthill IC, Emerson M, Altman DG. Animal research: reporting in vivo experiments: the ARRIVE guidelines. Br J Pharmacol 2010; 160: 1577-1579.

Higgins JP, Thompson SG. Quantifying heterogeneity in a meta-analysis. Stat Med 2002; 21: 1539-1558.

Zintzaras E, Ioannidis JP. Heterogeneity testing in meta-analysis of genome searches. Genet Epidemiol 2005; 28: 123-137.

Jove M, Pamplona R, Prat J, Arola L, Portero-Otin M. Atherosclerosis prevention by nutritional factors: a meta-analysis in small animal models. Nutr Metab Cardiovasc Dis 2013; 23: 84-93.

Wen J, Li YP. The selection of a summary statistic for use in meta-analysis. Chin J Evid Based Med 2007; 7: 606-613.

Nian H, Ma MH, Nian SS, Xu LL. Antiosteoporotic activity of icariin in ovariectomized rats. Phytomedicine 2009; 16: 320-326.

Qin L, Han T, Zhang Q, Cao D, Nian H, Rahman K, et al. Antiosteoporotic chemical constituents from Er-Xian Decoction, a traditional Chinese herbal formula. J Ethnopharmacol 2008; 118: 271-279.

Yang L, Yu Z, Qu H, Li M. Comparative effects of hispidulin, genistein, and icariin with estrogen on bone tissue in ovariectomized rats. Cell Biochem Biophys 2014; 70: 485-490.

Liu M, Zhong C, He RX, Chen LF. Icariin associated with exercise therapy is an effective treatment for postmenopausal osteoporosis. Chin Med J 2012; 125: 1784-1789.

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: 1007-1016.

Bian Q, Liu SF, HUang JH, Yang Z, Ning Y, Zhao YJ, Wang YJ, Shen ZY. Regulation and control of three active components of TCM effective in reinforcing kidney on ovariectomy-induced osteoporosis rats's bone marrow stem cells revealed by gene microarray. Chin J TCM 2011; 26: 889-893.

Qian GF, Yang H, Zhang XZ. Effect of icariin on the expression of Cbfa1in the bone of ovariectomized rats. Chin J Osteoporosis & Bone Miner Res 2009; 2: 104-109.

Wu MS, Zhao SZ, Ren LZ, Wang R, Bai X. Effects of icariin on expression of the estrogen receptor beta mRNA of bone and different hypothalamic nuclei in ovariectomy rats. Chin Pharm Bulletin 2011; 27: 29-33.

Bao JR, Yang JW, Li SF, Zhao W, Zhang QL, Yan YQ. Effects of icarrin on ovariectomized rats. J Hygene Res 2005; 34: 191-193.

Song Y, Li SH, He ZJ, Zhang YW, Zhou MW. Effect of icariin on osteoporosis in ovariectomized female rats. J Chin PLA Postgard Med Sch 2012; 33: 400-403.

Bai J. Effects of icariin on ovariectomized osteoporotic rats [Dissertation]. Lanzhou (China): Lanzhou University of Technology 2010.

Cheng K. Comparative research of icariin and genistein of pharmacological activity in antiosteoporosis [Dissertation]. Lanzhou (China): Lanzhou University of Technology 2014.

Liu SJ, Mo XM, Li JP, Yang J, Zhang XJ. Effecs of icariin and possible mechanisms in ovariectomized rats. Chin J Basic Med in TCM 2012; 3: 271-275.

Xue L, Wang Y, Jiang Y, Han T, Nie Y, Zhao L, et al. Comparative effects of er-xian decoction, epimedium herbs, and icariin with estrogen on bone and reproductive tissue in ovariectomized rats. Evid Based Complement Alternat Med 2012; 2012: 241416.

Aswar UM, Mohan V, Bodhankar SL. Antiosteoporotic activity of phytoestrogen-rich fraction separated from ethanol extract of aerial parts of Cissus quadrangularis in ovariectomized rats. Indian J Pharmacol 2012; 44: 345-350.

Kalu DN. The ovariectomized rat model of postmenopausal bone loss. Bone Miner 1991; 15: 175-191.

Manolagas SC, Jilka RL. Bone marrow, cytokines, and bone remodeling. Emerging insights into the pathophysiology of osteoporosis. N Engl J Med 1995; 332: 305-311.

Lerner UH. Bone remodeling in post-menopausal osteoporosis. J Dent Res 2006; 85: 584-595.

Mukaiyama K, Kamimura M, Uchiyama S, Ikegami S, Nakamura Y, Kato H. Elevation of serum alkaline phosphatase (ALP) level in postmenopausal women is caused by high bone turnover. Aging Clin Exp Res 2015; 27: 413-418.

Yogesh HS, Chandrashekhar VM, Katti HR, Ganapaty S, Raghavendra HL, Gowda GK, et al. Anti-osteoporotic activity of aqueous-methanol extract of Berberis aristata in ovariectomized rats. J Ethnopharmacol 2011; 134: 334-338.

Hertrampf T, Schleipen B, Offermanns C, Velders M, Laudenbach U, Diel P. Comparison of the bone protective effects of an isoflavone-rich diet with dietary and subcutaneous administrations of genistein in ovariectomized rats. Toxicol Lett 2009; 184: 198-203.

Qian WQ, Yin H, Sun HT. Influence of different concentrationsof icariin on osteoblast metabolism of rats. Chin Med Herald 2011; 8: 23-25.

Guo HL, Zhao YF, Wang X, Xu Y, Zhan HS. Experimental study on the mechanism of icariin improving human osteoblasts proliferation and the expression of OPG protein. Chin J Orthop Trauma 2011; 24: 585-588.

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: 832-840.

Cao H, Ke Y, Zhang Y, Zhang CJ, Qian W, Zhang GL. Icariin stimulates MC3T3-E1 cell proliferation and differentiation through up-regulation of bone morphogenetic protein-2. Int J Mol Med 2012; 29: 435-439.

Zhao J, Ohba S, Komiyama Y, Shinkai M, Chung UI, Nagamune T. Icariin: a potential osteoinductive compound for bone tissue engineering. Tissue engineering Part A 2010; 16: 233-243.

Mok SK, Chen WF, Lai WP, Leung PC, Wang XL, Yao XS, et al. Icariin protects against bone loss induced by oestrogen deficiency and activates oestrogen receptor-dependent osteoblastic functions in UMR 106 cells. Br J Pharmacol 2010; 159: 939-949.

Zheng D, Peng S, Yang SH, Shao ZW, Yang C, Feng Y, et al. The beneficial effect of Icariin on bone is diminished in osteoprotegerin-deficient mice. Bone 2012; 51: 85-92.

Hsieh TP, Sheu SY, Sun JS, Chen MH, Liu MH. Icariin isolated from Epimedium pubescens regulates osteoblasts anabolism through BMP-2, SMAD4, and Cbfa1 expression. Phytomedicine 2010; 17: 414-423.

Liang W, Lin M, Li X, Li C, Gao B, Gan H, et al. Icariin promotes bone formation via the BMP-2/Smad4 signal transduction pathway in the hFOB 1.19 human osteoblastic cell line. Int J Mol Med 2012; 30: 889-895.

Bagi C, van der Meulen M, Brommage R, Rosen D, Sommer A. The effect of systemically administered rhIGF-I/IGFBP-3 complex on cortical bone strength and structure in ovariectomized rats. Bone 1995; 16: 559-565.

Jee WS, Yao W. Overview: animal models of osteopenia and osteoporosis. J Musculoskelet Neuronal Interact 2001; 1: 193-207.

Pound P, Ebrahim S, Sandercock P, Bracken MB, Roberts I. Where is the evidence that animal research benefits humans? BMJ 2004; 328: 514-517.

van der Worp HB, Howells DW, Sena ES, Porritt MJ, Rewell S, O'Collins V, et al. Can animal models of disease reliably inform human studies? PLoS Med 2010; 7: e1000245.

Ma HP, Ming LG, Ge BF, Zhai YK, Song P, Xian CJ, et al. Icariin is more potent than genistein in promoting osteoblast differentiation and mineralization in vitro. J Cell Biochem 2011; 112: 916-923.

Bebarta V, Luyten D, Heard K. Emergency medicine animal research: does use of randomization and blinding affect the results? Acad Emerg Med 2003; 10: 684-687.

Sterne JA, Sutton AJ, Ioannidis JP, Terrin N, Jones DR, Lau J, et al. Recommendations for examining and interpreting funnel plot asymmetry in meta-analyses of randomised controlled trials. BMJ 2011; 343: d4002.

Macleod MR, O'Collins T, Horky LL, Howells DW, Donnan GA. Systematic review and metaanalysis of the efficacy of FK506 in experimental stroke. J Cereb Blood Flow Metab 2005; 25: 713-721.

Author biographies is not available.