Search JIM Advanced Search

Journal of Chinese Integrative Medicine ›› 2009, Vol. 7 ›› Issue (4): 342-348.doi: 10.3736/jcim20090408

• Original Experimental Research • Previous Articles     Next Articles

Effects of berberine on differentiation and bone resorption of osteoclasts derived from rat bone marrow cells

WEI Peng,JIAO Lei,QIN Lu-ping,YAN Fei,HAN Ting,ZHANG Qiao-yan()   

  1. Department of Pharmacognosy, School of Pharmacy, Second Military Medical University, Shanghai 200433, China
  • Received:2008-10-30 Accepted:2009-01-02 Online:2009-04-20 Published:2018-10-15

Objective: To observe the effects of berberine on osteoclastic differentiation and bone resorption action in vitro, and to investigate the cellular mechanism of its inhibitory effects on bone resorption.
Methods: The multinucleated osteoclasts (MNCs) were derived by 1,25-dihydroxyvitamin D3 and dexamethasone from bone marrow cells in the coculture system with primary osteoblastic cells. The tartrate-resistant acid phosphatase (TRAP) staining and image analysis of bone resorption pit on dental slices were used to identify osteoclast. The activity of TRAP was measured by p-nitrophenyl sodium phosphate assay. The bone resorption pit area on the bone slices formed by osteoclasts was measured by computer image processing.
Results: At the concentrations of 0.1, 1 and 10 μmol/L, berberine dose-dependently suppressed the formation of TRAP-positive multinucleated cells, the TRAP activity and the osteoclastic bone resorption. The strongest inhibitory effect was exhibited at the concentration of 10 μmol/L, with the inhibiting rate of 60.45%, 42.12% and 72.69% respectively.
Conclusion: Berberine can decrease bone loss through inhibition of osteoclast formation, differentiation and bone resorption.

Key words: berberine, osteoporosis, osteoclasts, bone resorption

Figure 1

Osteoclasts stained by TRAP (Inverted phase contrast microscopy, ×400)"

Figure 2

Bone resorption pits formed by osteoclasts (Inverted phase contrast microscopy, ×100) A: Control; B: Dental slice cocultured with osteoclasts treated with 10 μmol/L berberine."

Figure 3

Osteoclast differentiation in morphology (Inverted phase contrast microscopy, ×200) A: Control, normal mature osteoclasts; B: Osteoclasts treated with berberine at 10 μmol/L."

Figure 4

Inhibitory effect of berberine on formation of the TRAP-positive multinucleated osteoclasts **P<0.01, vs control group. Data represent as x±s, n=15."

Figure 5

Inhibitory effect of berberine on the osteoclastic TRAP activity **P<0.01, vs control group. Data represent as x±s, n=15."

Figure 6

Inhibitory effect of berberine on bone resorption pit area formed by osteoclasts **P<0.01, vs control group. Data represent as x±s, n=15."

[1] Dai RC, Zhang L, Liao EY . Progress in diagnosis and treatment of osteoporosis[J]. Zhongguo Yi Kan, 2008,43(4):4-6
戴如春, 张丽, 廖二元 . 骨质疏松的诊治进展[J]. 中国医刊, 2008,43(4):4-6
[2] Davison S, Davis SR . Hormone replacement therapy: current controversies[J]. Clin Endocrinol (Oxf), 2003,58(3):249-261
doi: 10.1046/j.1365-2265.2003.01774.x
[3] Hu JP, Takahashi N, Yamada T . Coptidis rhizoma inhibits growth and proteases of oral bacteria[J]. Oral Dis, 2000,6(5):297-302
[4] Anis KV, Rajeshkumar NV, Kuttan R . Inhibition of chemical carcinogenesis by berberine in rats and mice[J]. Pharm Pharmacol, 2001,53(5):763-768
doi: 10.1211/0022357011775901
[5] Hwang JM, Kuo HC, Tseng TH, Liu JY, Chu CY . Berberine induces apoptosis through a mitochondria/caspases pathway in human hepatoma cells[J]. Arch Toxicol, 2006,80(2):62-73
doi: 10.1007/s00204-005-0014-8
[6] Kuo CL, Chou CC, Yung BY . Berberine complexes with DNA in the berberine-induced apoptosis in human leukemic HL-60 cells. Cancer Lett[J]. Cancer Lett, 1995,93(2):193-200
doi: 10.1016/0304-3835(95)03809-B
[7] Mantena SK, Sharma SD, Katiyar SK . Berberine, a natural product, induces G1-phase cell cycle arrest and caspase-3-dependent apoptosis in human prostate carcinoma cells[J]. Mol Cancer Ther, 2006,5(2):296-308
doi: 10.1158/1535-7163.MCT-05-0448
[8] Li H, Miyahara T, Tezuka Y, Namba T, Nemoto N, Tonami S, Seto H, Tada T, Kadota S . The effect of Kampo formulae on bone resorption in vitro and in vivo. Ⅰ. Active constituents of Tsu-kan-gan[J]. Biol Pharm Bull, 1998,21(12):1322-1326
doi: 10.1248/bpb.21.1322
[9] Li H, Miyahara T, Tezuka Y, Namba T, Suzuki T, Dowaki R, Watanabe M, Nemoto N, Tonami S, Seto H, Kadota S . The effect of kampo formulae on bone resorption in vitro and in vivo. Ⅱ. Detailed study of berberine[J]. Biol Pharm Bull, 1999,22(4):391-396
doi: 10.1248/bpb.22.391
[10] Qin L, Han T, Zhang Q, Cao D, Nian H, Rahman K, Zheng H . Antiosteoporotic chemical constituents from Er-Xian Decoction, a traditional Chinese herbal formula[J]. J Ethnopharmacol, 2008,118(2):271-279
doi: 10.1016/j.jep.2008.04.009
[11] Nian H, Xu LL, Ma MH, Zheng HC, Zhang QY, Qin LP . Effects of berberine from Cortex Phellodendri Chinensis on osteoporosis in ovariectomized rats[J]. Yao Xue Fu Wu Yu Yan Jiu, 2007,7(1):41-44
年华, 徐玲玲, 马明华, 郑汉臣, 张巧艳, 秦路平 . 黄柏小檗碱对去卵巢大鼠骨质疏松症的作用[J]. 药学服务与研究, 2007,7(1):41-44
[12] Hu JP, Nishishita K, Sakai E, Yoshida H, Kato Y, Tsukuba T, Okamoto K . Berberine inhibits RANKL-induced osteoclast formation and survival through suppressing the NF-kappaB and Akt pathways[J]. Eur J Pharmacol, 2008,5802(1-2):70-79
[13] Rodan GA, Martin TJ . Role of osteoblasts in hormonal control of bone resorption — a hypojournal[J]. Calcif Tissue Int, 1981,33(4):349-351
doi: 10.1007/BF02409454
[14] Rodan GA, Martin TJ . Therapeutic approaches to bone diseases[J]. Science, 2000,289(5484):1508-1514
doi: 10.1126/science.289.5484.1508
[15] Teitelbaum SL . Bone resorption by osteoclasts[J]. Science, 2000,289(5484):1504-1508
doi: 10.1126/science.289.5484.1504
[16] Zhang QH, Gao HY, Wu LJ, Zhang L . Chemical constituents of Evodia rutaecarpa(Juss.) Benth[J]. Shenyang Yao Ke Da Xue Xue Bao, 2005,22(1):12-14
张起辉, 高慧媛, 吴立军, 张磊 . 吴茱萸的化学成分[J]. 沈阳药科大学学报, 2005,22(1):12-14
[17] Liu ZD, Zang HS, Ouyang YP . Biological study of growth pattern of newborn rat calvaria osteoblastic cell in vitro[J]. Jie Pou Xue Bao, 1995,26(2):157-160
刘祖德, 臧鸿声, 欧阳跃平 . 新生大鼠颅骨成骨性细胞体外生长过程研究[J]. 解剖学报, 1995,26(2):157-160
[18] Li B, Yu S . Genistein prevents bone resorption diseases by inhibiting bone resorption and stimulating bone formation[J]. Biol Pharm Bull, 2003,26(6):780-786
doi: 10.1248/bpb.26.780
[19] Yasukawa K, Takido M, Ikekawa T, Shimada F, Takeuchi M, Nakagawa S . Relative inhibitory activity of berberine-type alkaloids against 12-O-tetradecanoylphorbol-13-acetate-induced inflammation in mice[J]. Chem Pharm Bull (Tokyo), 1991,39(6):1462-1465
doi: 10.1248/cpb.39.1462
[20] Fukuda K, Hibiya Y, Mutoh M, Koshiji M, Akao S, Fujiwara H . Inhibition by berberine of cyclooxygenase-2 transcriptional activity in human colon cancer cells[J]. J Ethnopharmacol, 1999,66(2):227-233
doi: 10.1016/S0378-8741(98)00162-7
[21] Liu XH, Kirschenbaum A, Yao S, Levine AC . Cross-talk between the interleukin-6 and prostaglandin E2 signaling systems results in enhancement of osteoclastogenesis through effects on the osteoprotegerin/receptor activator of nuclear factor-kappaB(RANK) ligand/RANK system[J]. Endocrinology, 2005,146(4):227-233
[22] Zhang X, Schwarz EM, Young DA, Puzas JE, Rosier RN , O'Keefe RJ.Cyclooxygenase-2 regulates mesenchymal cell differentiation into the osteoblast lineage and is critically involved in bone repair[J]. J Clin Invest, 2002,109(11):1405-1415
doi: 10.1172/JCI0215681
[23] Ono K, Akatsu T, Murakami T, Kitamura R, Yamamoto M, Shinomiya N, Rokutanda M, Sasaki T, Amizuka N, Ozawa H, Nagata N, Kugai N . Involvement of cyclo-oxygenase-2 in osteoclast formation and bone destruction in bone metastasis of mammary carcinoma cell lines[J]. J Bone Miner Res, 2002,17(5):774-781
doi: 10.1359/jbmr.2002.17.5.774
[24] Zhang X, Morham SG, Langenbach R, Young DA, Xing L, Boyce BF, Puzas EJ, Rosier RN , O'Keefe RJ, Schwarz EM.Evidence for a direct role of cyclo-oxygenase 2 in implant wear debris-induced osteolysis[J]. J Bone Miner Res, 2001,16(4):660-670
doi: 10.1359/jbmr.2001.16.4.660
[1] Joon-ho Lee, Yuan-ji Wei, Zhong-yan Zhou, Yu-ming Hou, Cheng-long Wang, Li-bo Wang, Hong-jin Wu, Yu Zhang, Wei-wei Dai. Efficacy of the herbal pair, Radix Achyranthis Bidentatae and Eucommiae Cortex, in preventing glucocorticoid-induced osteoporosis in the zebrafish model. Journal of Integrative Medicine, 2022, 20(1): 83-90.
[2] Mohamed S. Othman, Sofian T. Obeidat, Amal H. Al-Bagawi, Mohamed A. Fareid, Alaa Fehaid, Ahmed E. Abdel Moneim. Green-synthetized selenium nanoparticles using berberine as a promising anticancer agent#br#
. Journal of Integrative Medicine, 2022, 20(1): 65-72.
[3] Meng-ya Shan, Ying Dai, Xiao-dan Ren, Jing Zheng, Ke-bin Zhang, Bin Chen, Jun Yan, Zi-hui Xu. Berberine mitigates nonalcoholic hepatic steatosis by downregulating SIRT1-FoxO1-SREBP2 pathway for cholesterol synthesis. Journal of Integrative Medicine, 2021, 19(6): 545-554.
[4] Hua Zhang, Xiang-yang Yao, Dan-feng Zhang, Qi-le Guo, Jun-jun Yang, Bao-guo Zhu, Dong-xiao Shen, Ke-feng Ruan, Yuan Wang, Jun-liang Peng. Anti-hemorrhoidal activity of Lian-Zhi-San, a traditional Chinese medicine, in an experimental hemorrhoidal model in rats. Journal of Integrative Medicine, 2021, 19(1): 42-49.
[5] Noriko Shinjyo, James Parkinson, Jimmy Bell, Tatsuro Katsuno, Annie Bligh. Berberine for prevention of dementia associated with diabetes and its comorbidities: A systematic review. Journal of Integrative Medicine, 2020, 18(2): 125-151.
[6] Syed Nasir Abbas Bukhari, Fahad Hussain, Hnin Ei Thu, Zahid Hussain. Synergistic effects of combined therapy of curcumin and Fructus Ligustri Lucidi for treatment of osteoporosis: cellular and molecular evidence of enhanced bone formation. Journal of Integrative Medicine, 2019, 17(1): 38-45.
[7] Lei Liu, Yuan-hui Guo , Hai-liang Xin , Yan Nie , Ting Han, Lu-ping Qin , Oiao-yan Zhang. Antiosteoporotic effects of benzylbenzoate glucosides from Curculigo orchioides in ovariectomized rats. Journal of Chinese Integrative Medicine, 2012, 10(12): 1419-1426.
[8] Gui-lin Ouyang, Xiao-hui Feng, Lian-bo Xiao, Zheng Huang, Qing Xia, Da-peng Han, Fei Zhu . Effects of Chinese herbal medicine Qianggu Capsule on patients with rheumatoid arthritis-induced osteoporosis: a report of 82 cases. Journal of Chinese Integrative Medicine, 2012, 10(12): 1394-1399.
[9] Xiang-ying Mao , Qin Bian , Zi-yin Shen. Analysis of the osteogenetic effects exerted on mesenchymal stem cell strain C3H10T1/2 by icariin via MAPK signaling pathway in vitro. Journal of Chinese Integrative Medicine, 2012, 10(11): 1272-1278.
[10] Dong-tao Li, Jian Wang , Hong-yang Jiang , Feng-lei Shi , Fu-yu Li , Ji-hong Liu , Yong-mei Cheng , Nan Yan , Ai-hua Hu, Mei-zeng Zhang , Jie Li, Ling-bo Wei , Rong-qin Jiang. Quantitative evaluation of the degrees of traditional Chinese medicine qualitative syndromes of osteoporosis. Journal of Chinese Integrative Medicine, 2012, 10(11): 1254-1262.
[11] Lei Bao, Shi-en Zou, Shao-fen Zhang. Dose-dependent effects of daidzein in regulating bone formation through estrogen receptors and peroxisome proliferator-activated receptor γ. Journal of Chinese Integrative Medicine, 2011, 9(2): 165-172.
[12] Zhi-ming Huang, Gui-ling Ouyang, Lian-bao Xiao, Ning-li Li, Hua-li Gao, Yong He, Zheng Huang, Xin-xing Huang. Effects of Drynaria total flavonoids on apoptosis of osteoblasts mediated by tumor necrosis factor-α. Journal of Chinese Integrative Medicine, 2011, 9(2): 173-178.
[13] Li Dong-tao, Li Fu-yu, Wang Jian, Liu Ji-hong, Yan Nan, Cheng Yong-mei, Hu Ai-hua, Jiang Hong-yang, Shi Feng-lei, Zhang Mei-zeng, Li Jie, Wei Ling-bo, Jiang Rong-qin. A study of diagnostic criteria for traditional Chinese medicine syndromes in osteoporosis. Journal of Chinese Integrative Medicine, 2011, 9(12): 1326-1332.
[14] Praveen Srikanta, Shivaprasad H. Nagarajappa, Gollapalle L. Viswanatha, Mukund Handral, Rajesh Subbanna, Rangappa Srinath, Ganesh Hiremath. Anti-osteoporotic activity of methanolic extract of an Indian herbal formula NR/CAL/06 in ovariectomized rats. Journal of Chinese Integrative Medicine, 2011, 9(10): 1125-1132.
[15] Shu-qiang Wang, Wu Rao, Lian-fu Deng, Ya-ping Zhu, Yong-qiang Chen. Effects of rat serum containing Chinese herbal medicine Sangen Decoction on osteoclastogenesis and bone resorption of osteoclasts induced by polymethylmethacrylate particles. Journal of Chinese Integrative Medicine, 2011, 9(1): 64-69.
Full text



[1] Dong Yang, Yong-ping Du, Qing Shen, Wei Chen, Yan Yu, Guang-lei Chen. Expression of alpha-smooth muscle actin in renal tubulointerstitium in patients with kidney collateral stasis. Journal of Chinese Integrative Medicine, 2008, 6(1): 41-44
[2] Hai-feng Wei, Bai-liu Ya, Ling Zhao, Cui-fei Ye, Li Zhang, Lin Li. Evaluation of tongue manifestation of blood stasis syndrome and its relationship with blood rheological disorder in a rat model of transient brain ischemia. Journal of Chinese Integrative Medicine, 2008, 6(1): 73-76
[3] Xi Lin, Jian-ping Liu. Herbal medicines for viral myocarditis. Journal of Chinese Integrative Medicine, 2008, 6(1): 76
[4] Xi Lin, Jian-ping Liu. Tai chi for treating rheumatoid arthritis. Journal of Chinese Integrative Medicine, 2008, 6(1): 82
[5] Liang-ping Hu, Hui Gao. Discrimination of errors in statistical analysis of medical papers published in the first issue of 2006 in Journal of Chinese Integrative Medicine. Journal of Chinese Integrative Medicine, 2008, 6(1): 98-106
[6] Yan-bo Zhu , Qi Wang, Cheng-yu Wu, Guo-ming Pang, Jian-xiong Zhao, Shi-lin Shen, Zhong-yuan Xia , Xue Yan . Logistic regression analysis on relationships between traditional Chinese medicine constitutional types and overweight or obesity. Journal of Chinese Integrative Medicine, 2010, 8(11): 1023-1035
[7] Wei Xu, Meng Shi, Jian-gang Liu, Cheng-long Wang . Collagen protein expressions in ischemic myocardium of rats with acute myocardial infarction and effects of qi-tonifying, yin-tonifying and blood-activating herbs and detoxifying and blood-activating herbs. Journal of Chinese Integrative Medicine, 2010, 8(11): 1041-1047
[8] Tao Wang , Feng Qin. Effects of Chinese herbal medicine Xiaoyao Powder on monoamine neurotransmitters in hippocampus of rats with postpartum depression. Journal of Chinese Integrative Medicine, 2010, 8(11): 1075-1079
[9] Ying Xu , Chang-chun Zeng , Xiu-yu Cai , Rong-ping Guo , Guang Nie , Ying Jin. Chromaticity and optical spectrum colorimetry of the tongue color in different syndromes of primary hepatic carcinoma. Journal of Chinese Integrative Medicine, 2012, 10(11): 1263-1271
[10] Xiang-ying Mao , Qin Bian , Zi-yin Shen. Analysis of the osteogenetic effects exerted on mesenchymal stem cell strain C3H10T1/2 by icariin via MAPK signaling pathway in vitro. Journal of Chinese Integrative Medicine, 2012, 10(11): 1272-1278