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

作者：Wang, Qiang-Song; Zhang, Xin-Chang; Li, Rui-Xin; Sun, Jing-Gong; Su, Wei-Hua; Guo, Yong; Li, Hao; Zhang, Xi-Zheng

来源：BioMedical Engineering Online, 2015, 14(1): 46.

DOI：10.1186/s12938-015-0039-z

• Mechanical strain
• Icariin
• BMP-2
• NF-kappa B
• Bone remodeling

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-kappa B) signaling pathway was investigated with the expression of inhibitor of kappa B (I kappa B) a, phosphorylation of I kappa B-alpha (P-I kappa B-alpha), 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-kappa B signaling pathway by increasing the expression of I kappa B a, P-I kappa B-alpha, p65, P-p65 (P < 0.01). @@@ Conclusion: The combination of mechanical strain and icariin stimulation could activate the NF-kappa B pathway to improve the proliferation, differentiation of osteoblast-like cells.

• 出版日期2015-5-21
• 单位中国人民解放军军事医学科学院; 河北医科大学