Novel Soy Peptide CBP: Stimulation of Osteoblast Differentiation via TβRI-p38-MAPK-Depending RUNX2 Activation

Abstract

DEDEQIPSHPPR, the calcium-binding peptide (CBP) identified in soy yogurt, was proven to be a potential cofactor in osteoporosis prevention in our previous study, but the mechanism was unknown. In this study, the activity of alkaline phosphatase (ALP) and osteocalcin (OCN), the regulation of RUNX2, and the expression of TβRI were investigated to elucidate the underlying mechanism. The results show that CBP upregulated ALP activity and OCN concentration and increased the expression of RUNX2 and the activation of the MAPK signaling pathway. Similarly, the expression of osteogenesis-related genes in osteoblasts also increased upon CBP treatment. Moreover, the CBP-induced enhancement of ALP activity and phosphorylation levels in the p38 pathway was inhibited by treatment with a p38 inhibitor (SB203538) and TβRI inhibitor (SB431542), respectively, suggesting that p38 and TβRI were involved in the osteogenic action. Based on the signaling pathways, the intracellular calcium concentration was significantly elevated by CBP, which was correlated with the increased behavioral functions and the relative fluorescence intensity of the bone mass. These findings suggest that CBP stimulates osteoblast differentiation and bone mineralization through the activation of RUNX2 via mechanisms related to the TβRI-p38-MAPK signaling pathways, further highlighting CBP’s important potential for treating osteoporosis.

Calcium Binding Mechanism of Soybean Peptide with Histidine Alteration by Molecular Docking Analysis and Spectroscopic Methods

“…As the second messenger, calcium plays a crucial role in the process of cell differentiation. In previous studies, it was found that CBP can promote intracellular calcium concentration and thus promote the formation of new bone [ 17 ]. To further explore the effect of CBP-H on osteoblasts, flow cytometry analysis was conducted.…”

“…For example, Huang et al isolated and purified a high calcium affinity peptide Thr-Cys-His from shrimp [ 16 ]. Recently, we purified a novel soy peptide CBP (DEDEQIPSHPPR) with a high calcium affinity and osteogenesis activity, which has one His residue [ 17 ]. However, the influence of histidine on the interaction between peptides and calcium is still unclear.…”

Fabrication of Low-Molecular-Weight Hyaluronic Acid–Carboxymethyl Cellulose Hybrid to Promote Bone Growth in Guided Bone Regeneration Surgery: An Animal Study

“…Because the L-HLA-induced cell viability was significantly reduced by the MAPK inhibitors, it is reasonable to suggest that MAPK signaling pathway activation is responsible for enhanced L-HLA-induced DPSC proliferation. It is well-known that the expression of various osteogenesis-related markers such as ALP, RUNX-2, and OCN can be affected by MAPK pathway regulation [ 45 ]. However, these markers may be down-regulated, by H-HLA in particular, at higher concentrations [ 46 ].…”

Isolation, characterization, and molecular docking analyses of novel calcium‐chelating peptide from soy yogurt and the study of its calcium chelation mechanism

“…As zebrafish bones generally take 3 days to develop, suitable 3‐day post‐fertilization (dpf) embryos were selected for the experiment. The mean number of embryos produced was 200–300 and these were kept in fish water at a temperature of 28.5 °C (2 mg mL −1 instant ocean salt) 29 …”

“…The mean number of embryos produced was 200-300 and these were kept in fish water at a temperature of 28.5 °C (2 mg mL −1 instant ocean salt). 29 Experimental design using zebrafish embryos A zebrafish model of osteoporosis was constructed according to the method of Yan et al 30 For the experiments, 3 dpf zebrafish embryos (10 embryos each) were prepared in a six-well plate and divided into different groups. Briefly, all zebrafish were directly immersed in prednisolone (2.5 μ mol L −1 , Solarbio, Beijing, China).…”