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Background: Icariin, a major active flavonoid glucoside, is widely used for the treatment of bone injury and rebuilding in the clinic because of its roles in suppressing osteoblastogenesis and promoting osteogenesis. The senescence-accelerated mouse SAMP6 was accepted as a useful murine model to reveal the mechanism of senile osteoporosis and the therapeutic mechanism of drug activity. However, little is known about the characteristics of SAMP6 osteoblasts and the associated regulatory roles of icariin.

Methods: We isolated and cultured osteoblasts from SAMP6 or SAMR1 mice and compared their proliferation, migration, and differentiation by performing the CCK-8 assay, cell counting assay, EdU staining, cell cycle analysis, ALP staining and activity measurement, Alizarin red staining, and RT-qPCR analysis to measure the levels of osteoblast markers, including RUNX2, Colla1 and Oc. To assess the effects of icariin on BMP-2-induced osteoblast differentiation, after BMP-2 treatment, osteoblast markers were analyzed by RT-qPCR and semi-quantitative Western blotting. The effects of icariin on connective tissue growth factor (CTGF) were measured by RT-qPCR. shRNA targeting CTGF mRNA was employed to knockdown its expression level in osteoblasts.

Results: The SAMP6 osteoblasts presented decreased the development and differentiation activity compared with SAMR1 osteoblasts, indicating that they are the potential mechanisms underlying age-associated disease. Moreover, SAMP6 osteoblasts presented upregulated CTGF compared with SAMR1 osteoblasts. Icariin enhanced BMP-2-induced osteoblast differentiation by downregulating CTGF expression, which tightly regulates osteoblast differentiation. By downregulating CTGF, icariin treatment upregulated phosphate-Smad1/5/8, indicating its activating effects on the BMP signaling pathway.

Conclusion: These results suggest that decreased osteoblast development and function potentially contributes to age-associated disease. Icariin exerts enhancing effects on BMP-2-mediated osteoblast development via downregulating CTGF.

Competing Interests: The authors have declared that no competing interests exist.

Titel:	Flavonoid compound icariin enhances BMP-2 induced differentiation and signalling by targeting to connective tissue growth factor (CTGF) in SAMP6 osteoblasts.
Autor/in / Beteiligte Person:	Xu, B ; Wang, X ; Wu, C ; Zhu, L ; Chen, O
Link:	Zum Volltext
Zeitschrift:	PloS one, Jg. 13 (2018-07-10), Heft 7, S. e0200367
Veröffentlichung:	San Francisco, CA : Public Library of Science, 2018
Medientyp:	academicJournal
ISSN:	1932-6203 (electronic)
DOI:	10.1371/journal.pone.0200367
Schlagwort:	Animals Bone Morphogenetic Protein 2 administration & dosage Cell Movement drug effects Cell Movement physiology Cell Proliferation drug effects Cell Proliferation physiology Cells, Cultured Disease Models, Animal Osteoblasts metabolism Osteoblasts pathology Osteogenesis drug effects Osteogenesis physiology Osteoporosis metabolism Osteoporosis pathology RNA, Messenger metabolism Recombinant Proteins administration & dosage Recombinant Proteins metabolism Signal Transduction drug effects Species Specificity Bone Morphogenetic Protein 2 metabolism Connective Tissue Growth Factor metabolism Flavonoids pharmacology Osteoblasts drug effects Osteoporosis drug therapy Protective Agents pharmacology
Sonstiges:	Nachgewiesen in: MEDLINE Sprachen: English Publication Type: Journal Article; Research Support, Non-U.S. Gov't Language: English [PLoS One] 2018 Jul 10; Vol. 13 (7), pp. e0200367. <i>Date of Electronic Publication: </i>2018 Jul 10 (<i>Print Publication: </i>2018). MeSH Terms: Bone Morphogenetic Protein 2 / metabolism ; Connective Tissue Growth Factor / metabolism ; Flavonoids / pharmacology ; Osteoblasts / drug effects ; Osteoporosis / drug therapy ; Protective Agents / pharmacology ; Animals ; Bone Morphogenetic Protein 2 / administration & dosage ; Cell Movement / drug effects ; Cell Movement / physiology ; Cell Proliferation / drug effects ; Cell Proliferation / physiology ; Cells, Cultured ; Disease Models, Animal ; Osteoblasts / metabolism ; Osteoblasts / pathology ; Osteogenesis / drug effects ; Osteogenesis / physiology ; Osteoporosis / metabolism ; Osteoporosis / pathology ; RNA, Messenger / metabolism ; Recombinant Proteins / administration & dosage ; Recombinant Proteins / metabolism ; Signal Transduction / drug effects ; Species Specificity References: Histol Histopathol. 2004 Jul;19(3):677-85. (PMID: 15168327) ; J Bone Miner Res. 2006 Jan;21(1):78-88. (PMID: 16355276) ; Dev Dyn. 2012 Dec;241(12):1944-59. (PMID: 23073844) ; Br J Pharmacol. 2010 Feb;159(4):939-49. (PMID: 20128811) ; Phytomedicine. 2010 May;17(6):414-23. (PMID: 19747809) ; Neurobiol Aging. 1998 Jan-Feb;19(1):71-6. (PMID: 9562506) ; J Cell Physiol. 2013 Mar;228(3):513-21. (PMID: 22777826) ; Exp Gerontol. 1997 Jan-Apr;32(1-2):117-27. (PMID: 9088909) ; Bone. 2009 Jan;44(1):80-6. (PMID: 18848655) ; PLoS One. 2012;7(3):e34122. (PMID: 22479539) ; J Bone Miner Metab. 2004;22(3):207-14. (PMID: 15108062) ; J Cell Physiol. 2003 Jul;196(1):51-62. (PMID: 12767040) ; J Bone Miner Res. 2006 Apr;21(4):637-46. (PMID: 16598384) ; Am J Pathol. 1986 Nov;125(2):276-83. (PMID: 3789087) ; J Cell Physiol. 2014 May;229(5):672-81. (PMID: 24127409) ; Front Pharmacol. 2017 Oct 31;8:784. (PMID: 29163169) ; Eur J Pharmacol. 2013 Aug 15;714(1-3):15-22. (PMID: 23764463) ; Pharmazie. 2007 May;62(5):388-91. (PMID: 17557750) ; Endocrinology. 2008 Sep;149(9):4374-81. (PMID: 18535099) ; J Clin Invest. 1996 Apr 1;97(7):1732-40. (PMID: 8601639) ; Connect Tissue Res. 2010 Dec;51(6):497-509. (PMID: 20497028) ; Stem Cell Rev. 2011 Sep;7(3):608-15. (PMID: 21318597) ; Sci Rep. 2015 Aug 25;5:13418. (PMID: 26302868) ; Bone Miner. 1994 Mar;24(3):245-55. (PMID: 8019211) ; Phytomedicine. 2009 Apr;16(4):320-6. (PMID: 19269147) ; Biochem Biophys Res Commun. 2008 Feb 8;366(2):450-6. (PMID: 18067859) ; Cell. 1997 May 30;89(5):747-54. (PMID: 9182762) ; Exp Gerontol. 1997 Jan-Apr;32(1-2):129-38. (PMID: 9088910) ; J Bone Miner Res. 1997 Nov;12(11):1772-9. (PMID: 9383681) ; J Control Release. 2016 Mar 10;225:152-69. (PMID: 26805518) ; J Cell Biochem. 2016 Jun;117(6):1396-406. (PMID: 26552846) ; J Biol Chem. 2013 Nov 22;288(47):33530-41. (PMID: 24106268) ; Bone. 2014 Sep;66:189-98. (PMID: 24956021) ; J Bone Miner Res. 2005 Mar;20(3):419-27. (PMID: 15746986) ; Phytomedicine. 2011 Jan 15;18(2-3):176-85. (PMID: 20554188) ; Gene. 2005 Aug 29;357(1):1-8. (PMID: 16125875) ; J Bone Miner Res. 2014 Nov;29(11):2427-38. (PMID: 24839202) ; Exp Hematol. 2012 Jun;40(6):499-509. (PMID: 22326715) ; J Bone Miner Res. 2002 Sep;17(9):1597-603. (PMID: 12211429) ; Exp Gerontol. 1997 Jan-Apr;32(1-2):105-9. (PMID: 9088907) ; J Cell Biochem. 1999 Sep 1;74(3):357-71. (PMID: 10412038) Substance Nomenclature: 0 (Bone Morphogenetic Protein 2) ; 0 (Flavonoids) ; 0 (Protective Agents) ; 0 (RNA, Messenger) ; 0 (Recombinant Proteins) ; 139568-91-5 (Connective Tissue Growth Factor) ; VNM47R2QSQ (icariin) Entry Date(s): Date Created: 20180711 Date Completed: 20190122 Latest Revision: 20190122 Update Code: 20231215 PubMed Central ID: PMC6039035

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Flavonoid compound icariin enhances BMP-2 induced differentiation and signalling by targeting to connective tissue growth factor (CTGF) in SAMP6 osteoblasts.