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Baicalin augments the differentiation of osteoblasts via enhancement of microRNA-217

  • Qi Wang
  • Donglei Shi
  • Yuanyuan Geng
  • Qishan Huang
  • Longzhan XiangEmail author
Article

Abstract

Baicalin (BAI), a sort of flavonoid monomer, acquires from Scutellaria baicalensis Georgi, which was forcefully reported in diversified ailments due to the pleiotropic properties. But, the functions of BAI in osteoblast differentiation have not been addressed. The intentions of this study are to attest the influences of BAI in the differentiation of osteoblasts. MC3T3-E1 cells or rat primary osteoblasts were exposed to BAI, and then cell viability, ALP activity, mineralization process, and Runx2 and Ocn expression were appraised through implementing CCK-8, p-nitrophenyl phosphate (pNPP), Alizarin red staining, western blot, and RT-qPCR assays. The microRNA-217 (miR-217) expression was evaluated in MC3T3-E1 cells or rat primary osteoblasts after BAI disposition; meanwhile, the functions of miR-217 in BAI-administrated MC3T3-E1 cells were estimated after miR-217 inhibitor transfection. The impacts of BAI and miR-217 inhibition on Wnt/β-catenin and MEK/ERK pathways were probed to verify the involvements in BAI-regulated the differentiation of osteoblasts. BAI accelerated cell viability, osteoblast activity, and Runx2 and Ocn expression in MC3T3-E1 cells or rat primary osteoblasts, and the phenomena were mediated via activations of Wnt/β-catenin and MEK/ERK pathways. Elevation of miR-217 was observed in BAI-disposed MC3T3-E1 cells or rat primary osteoblasts, and miR-217 repression annulled the functions of BAI in MC3T3-E1 cell viability and differentiation. Additionally, the activations of Wnt/β-catenin and MEK/ERK pathways evoked by BAI were both restrained by repression of miR-217. These explorations uncovered that BAI augmented the differentiation of osteoblasts via activations of Wnt/β-catenin and MEK/ERK pathways by ascending miR-217 expression.

Keywords

Bone fracture Baicalin microRNA-217 Wnt/β-catenin MEK/ERK 

Notes

Acknowledgements

None.

Funding

None.

Compliance with ethical standards

Conflict of interest

The authors declare that they have no conflict of interest.

Supplementary material

11010_2019_3632_MOESM1_ESM.tif (642 kb)
Supplementary Figure 1 BAI expedited rat primary osteoblast viability and osteogenic differentiation. After BAI management (20, 50, and 100 μM), (A) CCK-8 assay for rat primary osteoblast viability evaluation; (B) p-nitrophenyl phosphate (pNPP) assay for ALP activity detection; (C) western blot assay for Runx2 and Ocn protein level estimation; (D and E) RT-qPCR assay for Runx2 and Ocn mRNA expression level determination. *P < 0.05, **P < 0.01, ***P < 0.001 (TIFF 641 kb)
11010_2019_3632_MOESM2_ESM.tif (253 kb)
Supplementary Figure 2 BAI promoted osteoblast activity. After BAI management (20, 50, and 100 μM, TRAP activity assay for mature osteoclasts evaluation in (A) MC3T3-E1 cells and (B) rat primary osteoblasts. **P < 0.01, ***P < 0.001 (TIFF 252 kb)
11010_2019_3632_MOESM3_ESM.tif (1.4 mb)
Supplementary Figure 3 BAI expedited the activations of Wnt/β-catenin and MEK/ERK pathways in rat primary osteoblasts. After BAI administration (20, 50, and 100 μM), (A and B) western blot assay for wnt3a and β-catenin protein level measurement; (C and D) western blot assay for p/t-MEK and p/t-ERK protein level determination. *P < 0.05, **P < 0.01 (TIFF 1437 kb)
11010_2019_3632_MOESM4_ESM.tif (171 kb)
Supplementary Figure 4 Enhancement of miR-217 was triggered by BAI in rat primary osteoblasts. After BAI management (20, 50, and 100 μM), RT-qPCR assay for the expression level of miR-217 evaluation in rat primary osteoblasts. *P < 0.05, **P < 0.01 (TIFF 171 kb)

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Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2019

Authors and Affiliations

  1. 1.Department of OrthopaedicsHeze Municipal HospitalHezeChina
  2. 2.Department of Comprehensive MedicalHeze Infectious Disease HospitalHezeChina
  3. 3.Department of OrthopaedicsThe Second Affiliated Hospital of Wenzhou Medical UniversityWenzhouChina

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