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Background: The combination of drugs and exercise was the effective treatment in bone injure and rebuilding in clinic. As mechanical strain has potential in inducing the differentiation of osteoblasts in our previous study, the further research to investigate the combination of mechanical strain and icariin stimulation on inducing osteoblast proliferation, differentiation and the possible mechanism in MC3T3-E1 cell line.

Methods: A whole cell enzyme-linked immunosorbent assay that detects the bromodeoxyuridine incorporation during DNA synthesis was applied to evaluate the proliferation. The mRNA expression of alkaline phosphatase (ALP), osteocalcin (OCN), type I collagen (Col I), bone morphogenetic protein-2 (BMP-2) and BMP-4 was detected by real-time reverse-transcription polymerase chain reaction. The activity of ALP was analyzed by ELISA and the protein expression of OCN, Col I and BMP-2 was assessed by western blot. Moreover, the activity of nuclear transcription factor kappa-B (NF-κB) signaling pathway was investigated with the expression of inhibitor of κB (IκB) α, phosphorylation of IκB-α (P-IκB-α), p65, P-p65 by western blot.

Results: We observed that compared to single mechanical strain or icariin stimulation, the mRNA and protein expressions of ALP (P < 0.05 or P < 0.01), OCN (P < 0.01) and Col I (P < 0.05 or P < 0.01) were increased significantly by the combination of mechanical strain and icariin stimulation. Moreover, the combination of mechanical strain and icariin stimulation could up-regulate the expression of BMP-2 (P < 0.01) and BMP-4 compared to single mechanical strain or icariin stimulation. The combination of mechanical strain and icariin stimulation could activate NF-κB signaling pathway by increasing the expression of IκB α, P-IκB-α, p65, P-p65 (P < 0.01).

Conclusion: The combination of mechanical strain and icariin stimulation could activate the NF-κB pathway to improve the proliferation, differentiation of osteoblast-like cells.

Titel:	A comparative study of mechanical strain, icariin and combination stimulations on improving osteoinductive potential via NF-kappaB activation in osteoblast-like cells.
Autor/in / Beteiligte Person:	Wang, QS ; Zhang, XC ; Li, RX ; Sun, JG ; Su, WH ; Guo, Y ; Li, H ; Zhang, XZ
Link:	Zum Volltext
Zeitschrift:	Biomedical engineering online, Jg. 14 (2015-05-21), S. 46
Veröffentlichung:	London : BioMed Central, [2002-, 2015
Medientyp:	academicJournal
ISSN:	1475-925X (electronic)
DOI:	10.1186/s12938-015-0039-z
Schlagwort:	3T3 Cells Alkaline Phosphatase metabolism Animals Bone Morphogenetic Protein 2 genetics Bone Morphogenetic Protein 2 metabolism Bone Morphogenetic Protein 4 genetics Bone Morphogenetic Protein 4 metabolism Bone Remodeling drug effects Cell Differentiation drug effects Cell Proliferation drug effects Collagen Type I genetics Collagen Type I metabolism Gene Expression Regulation drug effects Mice Osteoblasts metabolism Osteocalcin genetics Osteocalcin metabolism Signal Transduction drug effects Flavonoids pharmacology NF-kappa B metabolism Osteoblasts cytology Osteoblasts drug effects Stress, Mechanical
Sonstiges:	Nachgewiesen in: MEDLINE Sprachen: English Publication Type: Comparative Study; Journal Article; Research Support, Non-U.S. Gov't Language: English [Biomed Eng Online] 2015 May 21; Vol. 14, pp. 46. <i>Date of Electronic Publication: </i>2015 May 21. MeSH Terms: Stress, Mechanical* ; Flavonoids / pharmacology ; NF-kappa B / metabolism ; Osteoblasts / cytology ; Osteoblasts / drug effects ; 3T3 Cells ; Alkaline Phosphatase / metabolism ; Animals ; Bone Morphogenetic Protein 2 / genetics ; Bone Morphogenetic Protein 2 / metabolism ; Bone Morphogenetic Protein 4 / genetics ; Bone Morphogenetic Protein 4 / metabolism ; Bone Remodeling / drug effects ; Cell Differentiation / drug effects ; Cell Proliferation / drug effects ; Collagen Type I / genetics ; Collagen Type I / metabolism ; Gene Expression Regulation / drug effects ; Mice ; Osteoblasts / metabolism ; Osteocalcin / genetics ; Osteocalcin / metabolism ; Signal Transduction / drug effects References: Annu Rev Immunol. 1996;14:649-83. (PMID: 8717528) ; Bone. 1996 May;18(5):405-10. (PMID: 8739897) ; Cell. 1996 Oct 4;87(1):13-20. (PMID: 8858144) ; Am J Physiol. 1997 Sep;273(3 Pt 1):C810-5. (PMID: 9316399) ; Biomed Tech (Berl). 1997 Nov;42(11):305-9. (PMID: 9435145) ; Cytokine Growth Factor Rev. 2005 Jun;16(3):251-63. (PMID: 15871923) ; J Biomech. 2005 Sep;38(9):1838-45. (PMID: 16023471) ; Gene. 2006 Feb 15;367:1-16. (PMID: 16361069) ; J Clin Endocrinol Metab. 2006 May;91(5):1848-54. (PMID: 16449331) ; J Orthop Sports Phys Ther. 2010 Dec;40(12):834. (PMID: 21169721) ; Cell Physiol Biochem. 2010;26(6):1093-102. (PMID: 21220940) ; J Bone Miner Metab. 2011 Jul;29(4):466-76. (PMID: 21188609) ; PLoS One. 2012;7(3):e34122. (PMID: 22479539) ; J Ethnopharmacol. 2012 May 7;141(1):377-85. (PMID: 22414473) ; PLoS One. 2012;7(4):e35709. (PMID: 22539993) ; Ann Biomed Eng. 2012 Sep;40(9):1884-94. (PMID: 22441665) ; Biomed Eng Online. 2012;11:80. (PMID: 23098360) ; Eur J Pharmacol. 2013 Aug 15;714(1-3):15-22. (PMID: 23764463) ; J Bone Miner Res. 2000 Sep;15(9):1731-45. (PMID: 10976993) ; Biochem Biophys Res Commun. 2001 Jun 15;284(3):622-31. (PMID: 11396946) ; Mutat Res. 2001 Sep 1;480-481:243-68. (PMID: 11506818) ; Biol Reprod. 2002 Aug;67(2):668-73. (PMID: 12135912) ; Osteoporos Int. 2002 Sep;13(9):688-700. (PMID: 12195532) ; Endocr Rev. 2003 Apr;24(2):218-35. (PMID: 12700180) ; Biomaterials. 2003 Sep;24(21):3859-68. (PMID: 12818559) ; J Biomech. 2003 Aug;36(8):1087-96. (PMID: 12831733) ; Nat Med. 2006 Jun;12(6):612-3. (PMID: 16761003) ; J Surg Res. 2007 Dec;143(2):329-36. (PMID: 17950332) ; J Clin Invest. 1999 Dec;104(11):1603-12. (PMID: 10587524) ; J Biol Chem. 2008 Apr 25;283(17):11535-40. (PMID: 18296709) ; Phytomedicine. 2009 Apr;16(4):320-6. (PMID: 19269147) ; Bone. 2009 Aug;45(2):367-76. (PMID: 19414075) ; Chem Biol Interact. 2010 Mar 30;184(3):413-22. (PMID: 20004183) ; Br J Pharmacol. 2010 Feb;159(4):939-49. (PMID: 20128811) ; Connect Tissue Res. 2010 Dec;51(6):497-509. (PMID: 20497028) ; J Biomech. 2004 Jan;37(1):157-61. (PMID: 14672580) ; Med Biol Eng Comput. 2004 Jan;42(1):14-21. (PMID: 14977218) ; Sci STKE. 2004 Sep 7;2004(249):RE12. (PMID: 15353762) ; FASEB J. 2004 Oct;18(13):1524-35. (PMID: 15466361) ; Clin Orthop Relat Res. 1972 Sep;87:132-7. (PMID: 4263296) ; J Bone Joint Surg Am. 1984 Mar;66(3):397-402. (PMID: 6699056) ; Bone Miner. 1992 Dec;19(3):257-71. (PMID: 1472896) ; Proc Natl Acad Sci U S A. 1993 Mar 15;90(6):2532-6. (PMID: 8460169) ; Annu Rev Immunol. 1994;12:141-79. (PMID: 8011280) ; Ann Biomed Eng. 1994 Jan-Feb;22(1):14-22. (PMID: 8060022) ; J Bone Miner Res. 1995 May;10(5):683-9. (PMID: 7639102) Substance Nomenclature: 0 (Bmp2 protein, mouse) ; 0 (Bmp4 protein, mouse) ; 0 (Bone Morphogenetic Protein 2) ; 0 (Bone Morphogenetic Protein 4) ; 0 (Collagen Type I) ; 0 (Flavonoids) ; 0 (NF-kappa B) ; 104982-03-8 (Osteocalcin) ; EC 3.1.3.1 (Alkaline Phosphatase) ; VNM47R2QSQ (icariin) Entry Date(s): Date Created: 20150522 Date Completed: 20160217 Latest Revision: 20181113 Update Code: 20231215 PubMed Central ID: PMC4455701

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A comparative study of mechanical strain, icariin and combination stimulations on improving osteoinductive potential via NF-kappaB activation in osteoblast-like cells.