Green tea (Camellia sinensis) aqueous extract alleviates postmenopausal osteoporosis in ovariectomized rats and prevents RANKL-induced osteoclastogenesis in vitro

  • Xin Wu
  • Chuan-qi Xie
  • Qiang-qiang Zhu
  • Ming-yue Wang
  • Bin Sun
  • Yan-ping Huang
  • Chang Shen
  • Meng-fei An
  • Yun-li Zhao
  • Xuan-jun Wang
  • Jun Sheng
Keywords: Green tea aqueous extract, Osteoporosis, Ovariectomy, Receptor activator of the nuclear factor kappa B ligand, Osteoclast


Background: Green tea (Camelliasinensis [L.] Kuntze) belongs to the plant family Theaceae and is mainly distributed in East Asia, the Indian subcontinent and Southeast Asia. This plant has been proven to be beneficial to human health, and green tea is the second most consumed beverage in the world after water. However, until now, the effect of green tea aqueous extract (GTE) upon postmenopausal osteoporosis has remained unclear. In this study, we investigated the therapeutic effects of GTE on estrogen deficiency-induced osteoporosis and explored the possible mechanisms in vivo and in vitro.

Materials and methods: Ovariectomized (OVX) female rats were orally administered with GTE at doses of 60,
120, and 370 mg kg−1 for 13 consecutive weeks. The biochemical parameters, bone gla protein, alkaline phosphatase, acid phosphatase, estrogen, interleukin-1β, and interleukin-6 in blood samples were detected, and histological change in bones was analyzed by hematoxylin and eosin staining. Meanwhile, the mechanisms of GTE on osteoclast formation were explored in RAW 264.7 cells induced by receptor activation of the nuclear
factor kappa B ligand (RANKL).

Results: The results showed that GTE could increase bone mass and inhibit trabecular bone loss in OVX
rats. Furthermore, real-time quantitative reverse transcription polymerase chain reaction analysis from
in vitro experiments also showed that GTE reduced the mRNA expression of osteoclast-associated genes such
as cathepsin K (cath-K), c-Fos, matrix metalloproteinase 9, nuclear factor of activated T cells cytoplasmic 1
(NFATc1) and tartrate-resistant acid phosphatase. In addition, GTE caused a reduction in the protein levels of
NFATc1, c-Fos, c-src and cath-K.

Conclusion: Evidence from both animal models and in vitro experiments suggested that GTE might effectively
ameliorate the symptoms of osteoporosis in OVX rats and inhibit RANKL-induced osteoclast-specific gene and protein expression.


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  1. Nelson ER, Wardell SE, McDonnell DP. The molecular mechanisms underlying the pharmacological actions of estrogens, SERMs and oxysterols: implications for the treatment and prevention of osteoporosis. Bone 2013; 53(1): 42–50.

  2. Sharma C, Mansoori MN, Dixit M, Shukla P, Kumari T, Bhandari SP, et al. Ethanolic extract of Coelogyne cristata Lindley (Orchidaceae) and its compound coelogin promote osteoprotective activity in ovariectomized estrogen deficient mice. Phytomedicine 2014; 21(12): 1702–7.

  3. Li Y, Lü SS, Tang GY, Hou M, Tang Q, Zhang XN, et al. Effect of Morinda officinalis capsule on osteoporosis in ovariectomized rats. Chinese J Nat Med 2014; 12(3): 204–12.

  4. Casarrubios L, Matesanz MC, Sanchez-Salcedo S, Arcos D, Vallet-Regi M, Portoles MT. Nanocrystallinity effects on osteoblast and osteoclast response to silicon substituted hydroxyapatite. J Colloid Interface Sci 2016; 482: 112–20.

  5. Andersen CY, Kristensen SG. Novel use of the ovarian follicular pool to postpone menopause and delay osteoporosis. Reprod Biomed Online 2015; 31(2): 128–31.

  6. Xu F, Ding Y, Guo Y, Liu B, Kou Z, Xiao W, et al. Anti-osteoporosis effect of Epimedium via an estrogen-like mechanism based on a system-level approach. J Ethnopharmacol 2016; 177: 148–60.

  7. Rossouw JE, Prentice RL, Manson JE, Wu L, Barad D, Barnabei VM, et al. Postmenopausal hormone therapy and risk of cardiovascular disease by age and years since menopause. JAMA 2007; 297(13): 1465–77.

  8. Al-Anazi AF, Qureshi VF, Javaid K, Qureshi S. Preventive effects of phytoestrogens against postmenopausal osteoporosis as compared to the available therapeutic choices: an overview. J Nat Sci Biol Med 2011; 2(2): 154–63.

  9. Wang C, Yu C, Gu Y, Zhang L. Research progress of drugs for osteoporosis treatment. Chinese Sci Bull 2014; 59(13): 1209–14.

  10. Khosla S. Update on estrogens and the skeleton. J Clin Endocrinol Metab 2010; 95(8): 3569–77.

  11. Karvande A, Khedgikar V, Kushwaha P, Ahmad N, Kothari P, Verma A, et al. Heartwood extract from Dalbergia sissoo promotes fracture healing and its application in ovariectomy-induced osteoporotic rats. J Pharm Pharmacol 2017; 69(10): 1381–97.

  12. Zhang R, Pan YL, Hu SJ, Kong XH, Juan W, Mei QB. Effects of total lignans from Eucommia ulmoides barks prevent bone loss in vivo and in vitro. J Ethnopharmacol 2014; 155(1): 104–12.

  13. Byun MR, Sung MK, Kim AR, Lee CH, Jang EJ, Jeong MG, et al. (-)-Epicatechin gallate (ECG) stimulates osteoblast differentiation via Runt-related transcription factor 2 (RUNX2) and transcriptional coactivator with PDZ-binding motif (TAZ)-mediated transcriptional activation. J Biol Chem 2014; 289(14): 9926–35.

  14. Tsai CF, Hsu YW, Ting HC, Huang CF, Yen CC. The in vivo antioxidant and antifibrotic properties of green tea (Camellia sinensis, Theaceae). Food Chem 2013; 136(3–4): 1337–44.

  15. Chen CH, Kang L, Lin RW, Fu YC, Lin YS, Chang JK, et al. (-)-Epigallocatechin-3-gallate improves bone microarchitecture in ovariectomized rats. Menopause J North Am Menopause Soc 2013; 20(6): 687–94.

  16. Suliburska J, Bogdanski P, Szulinska M, Stepien M, Pupek-Musialik D, Jablecka A. Effects of green tea supplementation on elements, total antioxidants, lipids, and glucose values in the serum of obese patients. Biol Trace Elem Res 2012; 149(3): 315–22.

  17. Matoušková P, Bártíková H, Boušová I, Szotáková B, Martin J, Skorkovská J, et al. Effect of defined green tea extract in various dosage schemes on drug-metabolizing enzymes in mice in vivo. J Funct Foods 2014; 10: 327–35.

  18. Shen CL, Yeh JK, Cao JJ, Wang JS. Green tea and bone metabolism. Nutr Res 2009; 29(7): 437–56.

  19. Shen CL, Yeh JK, Cao JJ, Chyu MC, Wang JS. Green tea and bone health: evidence from laboratory studies. Pharmacol Res 2011; 64(2): 155–61.

  20. Oka Y, Iwai S, Amano H, Irie Y, Yatomi K, Ryu K, et al. Tea polyphenols inhibit rat osteoclast formation and differentiation. J Pharmacol Sci 2012; 118(1): 55–64.

  21. Shen CL, Wang P, Guerrieri J, Yeh JK, Wang JS. Protective effect of green tea polyphenols on bone loss in middle-aged female rats. Osteoporos Int 2008; 19(7): 979–90.

  22. Zhengyi PHR, Eds Wu, Peter H, Wu Z. Flora of China. Bei Jing: Science Press; 2000.

  23. Henning SM, Niu Y, Liu Y, Lee NH, Hara Y, Thames GD, et al. Bioavailability and antioxidant effect of epigallocatechin gallate administered in purified form versus as green tea extract in healthy individuals. J Nutr Biochem 2005; 16(10): 610–16.

  24. Chunxiao W, Chengying G, Liang J, Xiaoming S, Feng G, Junting Y, et al. Pharmacological effects of a recombinant hPTH(1-34) derived peptide on ovariectomized rats. Eur J Pharmacol 2017; 794: 193–200.

  25. Wang S, Huang Y, Xu H, Zhu Q, Lu H, Zhang M, et al. Oxidized tea polyphenols prevent lipid accumulation in liver and visceral white adipose tissue in rats. Eur J Nutr 2016; 56(6): 2037–48.

  26. Garcia Palacios V, Robinson LJ, Borysenko CW, Lehmann T, Kalla SE, Blair HC. Negative regulation of RANKL-induced osteoclastic differentiation in RAW264.7 cells by estrogen and phytoestrogens. J Biol Chem 2005; 280(14): 13720–7.

  27. Cai X, Fang C, Hayashi S, Hao S, Zhao M, Tsutsui H, et al. Pu-erh tea extract ameliorates high-fat diet-induced nonalcoholic steatohepatitis and insulin resistance by modulating hepatic IL-6/STAT3 signaling in mice. J Gastroenterol 2016; 51(8): 819–29.

  28. Badr El Dine FMM, Nabil IM, Dwedar FI. The effect of tributyltin on thyroid follicular cells of adult male albino rats and the possible protective role of green tea: a toxicological, histological and biochemical study. Egypt J Forensic Sci 2017; 7(1): 7.

  29. Wang C, Yu C, Gu Y, Zhang L. Research progress of drugs for osteoporosis treatment. Chinese Sci Bull 2014; 59(13): 49–54.

  30. Khaw KT. Epidemiology of coronary heart disease in women. Heart 2006; 92(Suppl_3): iii2–iii4.

  31. Davis SR, Castelo-Branco C, Chedraui P, Lumsden MA, Nappi RE, Shah D, et al. Understanding weight gain at menopause. Climacteric 2012; 15(5): 419–29.

  32. Zhao LJ, Liu YJ, Liu PY, Hamilton J, Recker RR, Deng HW. Relationship of obesity with osteoporosis. J Clin Endocrinol Metab 2007; 92(5): 1640–6.

  33. Goldberg G. Nutrition and bone. Womens Health Med 2004; 1(1): 25–9.

  34. Kum CJ, Kim EY, Kim JH, Lee B, Min JH, Heo J, et al. Cyperus Rotundus L. extract suppresses RANKL-induced osteoclastogenesis through NFATc1/c-fos downregulation and prevent bone loss in OVX-induced osteoporosis rat. J Ethnopharmacol 2017; 205: 186–94.

  35. Asagiri M, Sato K, Usami T, Ochi S, Nishina H, Yoshida H, et al. Autoamplification of NFATc1 expression determines its essential role in bone homeostasis. J Exp Med 2005; 202(9): 1261–9.

  36. Takayanagi H, Kim S, Koga T, Nishina H, Isshiki M, Yoshida H, et al. Induction and activation of the transcription factor NFATc1 (NFAT2) integrate RANKL signaling in terminal differentiation of osteoclasts. Dev Cell 2002; 3: 889–901.

  37. Saftig P, Hunziker E, Everts V, Jones S, Boyde A, Wehmeyer O, et al. Functions of Cathepsin K in bone resorption. Adv Exp Med Biol 2000; 477: 293–303.

  38. Khan SA, Priyamvada S, Farooq N, Khan S, Khan MW, Yusufi AN. Protective effect of green tea extract on gentamicin-induced nephrotoxicity and oxidative damage in rat kidney. Pharmacol Res 2009; 59(4): 254–62.

How to Cite
Wu X, Xie C- qi, Zhu Q- qiang, Wang M- yue, Sun B, Huang Y- ping, Shen C, An M- fei, Zhao Y- li, Wang X- jun, Sheng J. Green tea (<em>Camellia sinensis</em>) aqueous extract alleviates postmenopausal osteoporosis in ovariectomized rats and prevents RANKL-induced osteoclastogenesis <em>in vitro</em&gt;. fnr [Internet]. 2018Oct.8 [cited 2018Oct.21];620. Available from:
Original Articles