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Effects of Achyranthes bidentata alcohol on proliferation capacity of osteoblasts and miRNA in Runx2

  • Authors:
    • Shan Hua
    • Xuexue Zhang
  • View Affiliations

  • Published online on: July 2, 2019     https://doi.org/10.3892/etm.2019.7723
  • Pages: 1545-1550
  • Copyright: © Hua et al. This is an open access article distributed under the terms of Creative Commons Attribution License.

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Abstract

Achyranthes bidentata is a herbal plant commonly used in the treatment of osteoporosis and bone nonunion with traditional Chinese medicine. Achyranthes bidentata alcohol is a major component extracted from Achyranthes bidentata, which has been proved to be able to exert a variety of pharmacological effects, such as anti‑inflammation, antipyresis, anti‑rheumatism, diuresis and anti‑osteoporosis. Thirty male Sprague-Dawley rats aged 4 weeks were used in the experiment. All primary rat osteoblasts were cultured and amplified for further experiments. The osteoblasts were divided into six groups (5 rats in each group): the culture medium control group, the 25 µg/ml achyranthol group, the 50 µg/ml achyranthol group, the 100 µg/ml achyranthol group, 200 µg/ml achyranthol group, and the 25 µM PD98059+200 µg/ml achyranthol group. In this study, the effect of Achyranthes bidentata alcohol on the proliferation of osteoblasts was detected via methyl thiazolyl tetrazolium (MTT) assay. The effect of Achyranthes bidentata alcohol on the alkaline phosphatase (ALP) activity in osteoblasts was analyzed via ALP assay. The effect of Achyranthes bidentata alcohol on the expression of osteoblast marker gene, Runt‑related transcription factor 2 (Runx2), was detected via reverse transcription‑quantitative polymerase chain reaction (RT‑qPCR) and immunohistochemistry. Moreover, the phosphorylation or activation of extracellular signal‑regulated kinase (ERK) in osteoblasts induced by Achyranthes bidentata alcohol was analyzed using western blotting. Achyranthes bidentata alcohol increased cell proliferation in a dose‑dependent manner, increased the micro ribonucleic acid (miRNA) level in Runx2, enhanced the ALP activity in osteoblasts, and stimulated the activation of ERK (P<0.05). The expression of Runx2 with the inhibitor PD98059 was decreased significantly compared with that in the Achyranthes bidentata alcohol group (P<0.01). Immunohistochemical results manifested that the percentage of Runx2 positive cells in treated tissues was obviously higher than that in untreated tissues (P<0.01). Therefore, Achyranthes bidentata alcohol promotes the proliferation capacity of osteoblasts in a dose‑dependent manner, enhances the expression of miRNA in Runx2, and stimulates the osteogenic differentiation of osteoblasts through activating the ERK signal transduction pathway.

References

1 

Niu Y, Li Y, Huang H, Kong X, Zhang R, Liu L, Sun Y, Wang T and Mei Q: Asperosaponin VI, a saponin component from Dipsacus asper wall, induces osteoblast differentiation through bone morphogenetic protein-2/p38 and extracellular signal-regulated kinase 1/2 pathway. Phytother Res. 25:1700–1706. 2011. View Article : Google Scholar : PubMed/NCBI

2 

Licata AA: Discovery, clinical development, and therapeutic uses of bisphosphonates. Ann Pharmacother. 39:668–677. 2005. View Article : Google Scholar : PubMed/NCBI

3 

Zhang P, Dai KR, Yan SG, Yan WQ, Zhang C, Chen DQ, Xu B and Xu ZW: Effects of naringin on the proliferation and osteogenic differentiation of human bone mesenchymal stem cell. Eur J Pharmacol. 607:1–5. 2009. View Article : Google Scholar : PubMed/NCBI

4 

Berger I, Piecha G, Rabkin R, Kaya N, Geldyyev A, Sun D, Chen Y, Koleganova N and Gross ML: Growth hormone treatment prevents osteoporosis in uremic rats. Histol Histopathol. 22:1231–1239. 2007.PubMed/NCBI

5 

Lühe A, Künkele KP, Haiker M, Schad K, Zihlmann C, Bauss F, Suter L and Pfister T: Preclinical evidence for nitrogen-containing bisphosphonate inhibition of farnesyl diphosphate (FPP) synthase in the kidney: Implications for renal safety. Toxicol In Vitro. 22:899–909. 2008. View Article : Google Scholar : PubMed/NCBI

6 

Deng HB, Cui DP, Jiang JM, Feng YC, Cai NS and Li DD: Inhibiting effects of Achyranthes bidentata polysaccharide and Lycium barbarum polysaccharide on nonenzyme glycation in D-galactose induced mouse aging model. Biomed Environ Sci. 16:267–275. 2003.PubMed/NCBI

7 

Shen H, Yuan Y, Ding F, Liu J and Gu X: The protective effects of Achyranthes bidentata polypeptides against NMDA-induced cell apoptosis in cultured hippocampal neurons through differential modulation of NR2A- and NR2B-containing NMDA receptors. Brain Res Bull. 77:274–281. 2008. View Article : Google Scholar : PubMed/NCBI

8 

Jin LQ, Zheng ZJ, Peng Y, Li WX, Chen XM and Lu JX: Opposite effects on tumor growth depending on dose of Achyranthes bidentata polysaccharides in C57BL/6 mice. Int Immunopharmacol. 7:568–577. 2007. View Article : Google Scholar : PubMed/NCBI

9 

Li JX, Hareyama T, Tezuka Y, Zhang Y, Miyahara T and Kadota S: Five new oleanolic acid glycosides from Achyranthes bidentata with inhibitory activity on osteoclast formation. Planta Med. 71:673–679. 2005. View Article : Google Scholar : PubMed/NCBI

10 

Wattel A, Kamel S, Prouillet C, Petit JP, Lorget F, Offord E and Brazier M: Flavonoid quercetin decreases osteoclastic differentiation induced by RANKL via a mechanism involving NF kappa B and AP-1. J Cell Biochem. 92:285–295. 2004. View Article : Google Scholar : PubMed/NCBI

11 

Shibeshi W, Makonnen E, Zerihun L and Debella A: Effect of Achyranthes aspera L. on fetal abortion, uterine and pituitary weights, serum lipids and hormones. Afr Health Sci. 6:108–112. 2006.PubMed/NCBI

12 

Chen XM, Xu YJ and Tian GY: Physical-chemical properties and structure elucidation of abPS isolated from the root of Achyranthes bidentata. Yao Xue Xue Bao. 40:32–35. 2005.(In Chinese). PubMed/NCBI

13 

Han SB, Lee CW, Yoon YD, Lee JH, Kang JS, Lee KH, Yoon WK, Lee K, Park SK and Kim HM: Prevention of arthritic inflammation using an oriental herbal combination BDX-1 isolated from Achyranthes bidentata and Atractylodes japonica. Arch Pharm Res. 28:902–908. 2005. View Article : Google Scholar : PubMed/NCBI

14 

Zhou ZD and Xia DJ: Effect of Achyranthes bidentata polysaccharides stimulated dendritic cells co-cultured with cytokine induced killer cells against SW480 cells. Zhongguo Zhong Yao Za Zhi. 38:1056–1060. 2013.(In Chinese). PubMed/NCBI

15 

Zhang M, Wang Y, Zhang Q, Wang C, Zhang D, Wan JB and Yan C: UPLC/Q-TOF-MS-based metabolomics study of the anti-osteoporosis effects of Achyranthes bidentata polysaccharides in ovariectomized rats. Int J Biol Macromol. 112:433–441. 2018. View Article : Google Scholar : PubMed/NCBI

16 

Zhang R, Hu SJ, Li C, Zhang F, Gan HQ and Mei QB: Achyranthes bidentata root extract prevent OVX-induced osteoporosis in rats. J Ethnopharmacol. 139:12–18. 2012. View Article : Google Scholar : PubMed/NCBI

17 

Pearson G, Robinson F, Beers Gibson T, Xu BE, Karandikar M, Berman K and Cobb MH: Mitogen-activated protein (MAP) kinase pathways: Regulation and physiological functions. Endocr Rev. 22:153–183. 2001. View Article : Google Scholar : PubMed/NCBI

18 

Jaiswal RK, Jaiswal N, Bruder SP, Mbalaviele G, Marshak DR and Pittenger MF: Adult human mesenchymal stem cell differentiation to the osteogenic or adipogenic lineage is regulated by mitogen-activated protein kinase. J Biol Chem. 275:9645–9652. 2000. View Article : Google Scholar : PubMed/NCBI

19 

Sale EM, Atkinson PG and Sale GJ: Requirement of MAP kinase for differentiation of fibroblasts to adipocytes, for insulin activation of p90 S6 kinase and for insulin or serum stimulation of DNA synthesis. EMBO J. 14:674–684. 1995. View Article : Google Scholar : PubMed/NCBI

20 

Alanay Y, Avaygan H, Camacho N, Utine GE, Boduroglu K, Aktas D, Alikasifoglu M, Tuncbilek E, Orhan D, Bakar FT, et al: Mutations in the gene encoding the RER protein FKBP65 cause autosomal-recessive osteogenesis imperfecta. Am J Hum Genet. 86:551–559. 2010. View Article : Google Scholar : PubMed/NCBI

21 

Wei J, Shimazu J, Makinistoglu MP, Maurizi A, Kajimura D, Zong H, Takarada T, Lezaki T, Pessin JE, Hinoi E, et al: Glucose uptake and Runx2 synergize to orchestrate osteoblast differentiation and bone formation. Cell. 161:1576–1591. 2015. View Article : Google Scholar : PubMed/NCBI

22 

Livak KJ and Schmittgen TD: Analysis of relative gene expression data using real-time quantitative PCR and the 2(-Delta Delta C(T)) method. Methods. 25:402–408. 2001. View Article : Google Scholar : PubMed/NCBI

23 

Nishio Y, Dong Y, Paris M, O'Keefe RJ, Schwarz EM and Drissi H: Runx2-mediated regulation of the zinc finger Osterix/Sp7 gene. Gene. 372:62–70. 2006. View Article : Google Scholar : PubMed/NCBI

24 

Komori T, Yagi H, Nomura S, Yamaguchi A, Sasaki K, Deguchi K, Shimizu Y, Bronson RT, Gao YH, Inada M, et al: Targeted disruption of Cbfa1 results in a complete lack of bone formation owing to maturational arrest of osteoblasts. Cell. 89:755–764. 1997. View Article : Google Scholar : PubMed/NCBI

25 

Nakashima K, Zhou X, Kunkel G, Zhang Z, Deng JM, Behringer RR and de Crombrugghe B: The novel zinc finger-containing transcription factor osterix is required for osteoblast differentiation and bone formation. Cell. 108:17–29. 2002. View Article : Google Scholar : PubMed/NCBI

26 

Lee MH, Kim YJ, Kim HJ, Park HD, Kang AR, Kyung HM, Sung JH, Wozney JM, Kim HJ and Ryoo HM: BMP-2-induced Runx2 expression is mediated by Dlx5, and TGF-beta 1 opposes the BMP-2-induced osteoblast differentiation by suppression of Dlx5 expression. J Biol Chem. 278:34387–34394. 2003. View Article : Google Scholar : PubMed/NCBI

27 

Turjanski AG, Vaqué JP and Gutkind JS: MAP kinases and the control of nuclear events. Oncogene. 26:3240–3253. 2007. View Article : Google Scholar : PubMed/NCBI

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September 2019
Volume 18 Issue 3

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Copy and paste a formatted citation
APA
Hua, S., & Hua, S. (2019). Effects of Achyranthes bidentata alcohol on proliferation capacity of osteoblasts and miRNA in Runx2. Experimental and Therapeutic Medicine, 18, 1545-1550. https://doi.org/10.3892/etm.2019.7723
MLA
Hua, S., Zhang, X."Effects of Achyranthes bidentata alcohol on proliferation capacity of osteoblasts and miRNA in Runx2". Experimental and Therapeutic Medicine 18.3 (2019): 1545-1550.
Chicago
Hua, S., Zhang, X."Effects of Achyranthes bidentata alcohol on proliferation capacity of osteoblasts and miRNA in Runx2". Experimental and Therapeutic Medicine 18, no. 3 (2019): 1545-1550. https://doi.org/10.3892/etm.2019.7723