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Vitexin enhances osteoblast differentiation through phosphorylation of Smad and expression of Runx2 at in vitro and ex vivo

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Abstract

Vitexin (apigenin-8-C-d-glucopyranoside) is a flavonoid isolated from natural sources. It has been employed as an anti-oxidant, anti-inflammatory, and anti-cancer agent, and is used as a traditional Chinese medicine to treat a variety of illnesses. The present study investigated the effect of vitexin on osteoblast differentiation of C3H10T1/2 mesenchymal stem cells, MC3T3-E1 preosteoblast, mouse calvarial primary cells, and primary bone marrow stem cells (BMSCs). RT-PCR and quantitative PCR demonstrated that vitexin increased mRNA expression of the osteogenic genes distal-less homeobox 5 (Dlx5) and Runxt-related transcription factor 2 (Runx2). Vitexin also increased the Dlx5 and Runx2 protein levels, Smad1/5/9 phosphorylation, and alkaline phosphatase (ALP) activity. In addition, vitexin increased Runx2-luciferase activity. Moreover, knockdown of Runx2 attenuated the increase in ALP activity induced by vitexin. These results demonstrate that vitexin enhances osteoblast differentiation via Runx2.

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Abbreviations

MSC:

Mesenchymal stem cell

BMSC:

Bone marrow stromal cell

BMP:

Bone morphogenetic proteins

Runx2:

Runt-related transcription factor 2

TNF-α:

Tumour necrosis factor-α

ALP:

Alkaline phosphatase

DMEM:

Dulbecco's Modified Eagle's Medium

Dlx5:

Distal-less homeobox 5

DMSO:

Dimethyl sulfoxide

Smad:

Small mothers against

FBS:

Fetal bovine serum

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Acknowledgements

This research was supported by Daegu University Research Grant 2018.

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Authors and Affiliations

  1. Department of Biotechnology, School of Engineering, Daegu University, Gyeongbuk, 38453, Republic of Korea

    Kyeong-Min Kim, Hyo-Eun Son, Hyeon-Young Min & Won-Gu Jang

  2. Research a of Anti-Aging, Daegu University, Gyeongbuk, 38453, Republic of Korea

    Kyeong-Min Kim, Hyo-Eun Son, Hyeon-Young Min & Won-Gu Jang

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  1. Kyeong-Min Kim
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Correspondence to Won-Gu Jang.

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Kim, KM., Son, HE., Min, HY. et al. Vitexin enhances osteoblast differentiation through phosphorylation of Smad and expression of Runx2 at in vitro and ex vivo. Mol Biol Rep 47, 8809–8817 (2020). https://doi.org/10.1007/s11033-020-05929-y

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