Vitexin enhances osteoblast differentiation through phosphorylation of Smad and expression of Runx2 at in vitro and ex v
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ORIGINAL ARTICLE
Vitexin enhances osteoblast differentiation through phosphorylation of Smad and expression of Runx2 at in vitro and ex vivo Kyeong‑Min Kim1,2 · Hyo‑Eun Son1,2 · Hyeon‑Young Min1,2 · Won‑Gu Jang1,2 Received: 4 October 2019 / Accepted: 17 October 2020 © Springer Nature B.V. 2020
Abstract Vitexin (apigenin-8-C-d-glucopyranoside) is a flavonoid isolated from natural sources. It has been employed as an antioxidant, 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. Keywords Vitexin · Osteogenic differentiation · Mesenchymal stem cell 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
Introduction Bone is a dynamic tissue that is remodeled via the actions of bone-forming osteoblasts and bone-resorbing osteoclasts [1]. An imbalance in these actions can lead to various diseases * Won‑Gu Jang [email protected] 1
Department of Biotechnology, School of Engineering, Daegu University, Gyeongbuk 38453, Republic of Korea
Research a of Anti‑Aging, Daegu University, Gyeongbuk 38453, Republic of Korea
2
such as osteoporosis, bone metastasis, and tumor-induced bone destruction, rheumatoid arthritis, and osteosclerosis [2, 3]. Therefore, bone remodeling is very important. Osteoblasts are derived from mesenchymal stem cells (MSCs), which are pluripotent cells that can differentiate into osteoblasts, adipocytes, myocytes, and chondrocyte [4–6]. Runx2 is a member of the runt domain gene family, also known as Cbfa1. Runx2 has first demonstrated in 1997 that the role of it was essential in osteoblast differentiation [7]. Runx2 role was important for the cell cycle and maturation stages during osteoblast differentiation [8]. Runx2 is a transcription factor that increases alkaline phosphatase (ALP) activity and matrix mineralization in vitro [9–11]. Previous reports indicate that Runx2 is a transcription factor integral to osteoblast differentiation. Vitexin (apigenin-8-C-d-glucopyranoside) is a f
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