“…In order to measure MC3T3‐E1 cells extracellular matrix calcium deposits, alizarin red S was applied to treat the cells. Details were from previous studies (Jie et al, 2018; Murshed, 2018; Saito et al, 2016). Briefly, cells were washed, fixed and then dyed with a 0.5% alizarin red S (pH 4.1).…”
Glucocorticoid‐induced osteoporosis (GIOP) that is mainly featured as low bone density and increased risk of fracture is prone to occur with the administration of excessive glucocorticoids. Cycloastragenol (CAG) has been verified to be a small molecule that activates telomerase. Studied showed that up‐regulated telomerase was associated with promoting osteogeneic differentiation, so we explored whether CAG could promote osteogenic differentiation to protect against GIOP and telomerase would be the target that CAG exerted its function. Our results demonstrated that CAG prominently increased the ALP activity, mineralization, mRNA of runt‐related transcription factor 2, osteocalcin, osteopontin, collagen type I in both MC3T3‐E1 cells and dexamethasone (DEX)‐treated MC3T3‐E1 cells. CAG up‐regulated telomerase reverse transcriptase and the protective effect of CAG was blocked by telomerase inhibitor TMPyP4. Moreover, CAG improved bone mineralization in DEX‐induced bone damage in a zebrafish larvea model. Therefore, the study showed that CAG could alleviate the osteogenic differentiation inhibition induced by DEX in vitro and in vivo, and CAG might be considered as a candidate drug for the treatment of GIOP.
“…In order to measure MC3T3‐E1 cells extracellular matrix calcium deposits, alizarin red S was applied to treat the cells. Details were from previous studies (Jie et al, 2018; Murshed, 2018; Saito et al, 2016). Briefly, cells were washed, fixed and then dyed with a 0.5% alizarin red S (pH 4.1).…”
Glucocorticoid‐induced osteoporosis (GIOP) that is mainly featured as low bone density and increased risk of fracture is prone to occur with the administration of excessive glucocorticoids. Cycloastragenol (CAG) has been verified to be a small molecule that activates telomerase. Studied showed that up‐regulated telomerase was associated with promoting osteogeneic differentiation, so we explored whether CAG could promote osteogenic differentiation to protect against GIOP and telomerase would be the target that CAG exerted its function. Our results demonstrated that CAG prominently increased the ALP activity, mineralization, mRNA of runt‐related transcription factor 2, osteocalcin, osteopontin, collagen type I in both MC3T3‐E1 cells and dexamethasone (DEX)‐treated MC3T3‐E1 cells. CAG up‐regulated telomerase reverse transcriptase and the protective effect of CAG was blocked by telomerase inhibitor TMPyP4. Moreover, CAG improved bone mineralization in DEX‐induced bone damage in a zebrafish larvea model. Therefore, the study showed that CAG could alleviate the osteogenic differentiation inhibition induced by DEX in vitro and in vivo, and CAG might be considered as a candidate drug for the treatment of GIOP.
“…Phosvitin likely mimics the role of ascorbic acid in osteoblast cell culture to promote bone mineralization [ 41 ]. These effects may be associated with its degree of phosphorylation, which can play a critical role in the proliferation and differentiation of osteoblast cells [ 42 ]. Since enzymatic hydrolysis to generate phospho-peptides would likely avoid the perceived pitfalls of native phosvitin, PPPs are a possible approach to enhancing osteogenic and/or osteoprotective functions.…”
Section: Discussionmentioning
confidence: 99%
“…Previous studies have also suggested that PPPs show enhanced bioactivities (anti-inflammation, antioxidant and calcium absorption promoting) compared to native phosvitin [ 43 , 44 ], further supporting the role(s) of phosphorylation in bioactive properties. In contrast, bioactive peptides lacking of phosphorylation, such as LRW derived from pea protein [ 45 ], VLPVPQK, EDVPSER, NAVPITPTL, HPHPHLSF derived from buffalo casein [ 46 ], YVEEL and YLLF derived from whey proteins [ 47 ], and IRW from egg proteins [ 48 ], were also reported to promote the proliferation of MC3T3-E1 cells, but they may not be as resistant to digestion as phospho-peptides, which therefore limits their potential applications [ 42 ]. Despite these potential benefits, the osteogenic capacity of PPP preparations is yet to be determined.…”
Phosphorylated proteins from food sources have been investigated as regulators of bone formation with potential benefits in treating osteoporosis. Egg, a cheap and nutritious food, is also the source of various proteins and bioactive peptides with applications in human health. Egg yolk is rich in phosvitin, the most phosphorylated protein in nature. Phosvitin has been shown to improve bone health in experimental animals, although the molecular mechanisms and its specific effects on bone-forming osteoblastic cells are incompletely understood. Previous work in our group has identified pancreatin-generated phosvitin phospho-peptides (PPP) as a potential source for bioactive peptides. Given this background, we examined the roles of both phosvitin and PPP in the function of osteoblastic cells. Our results demonstrated their potential to improve bone health by promoting osteoblast differentiation and proliferation, suppressing osteoclast recruitment and the deposition of extracellular matrix, although PPP appeared to demonstrate superior osteogenic functions compared to phosvitin alone.
“…The phosphorylated peptide-calcium complex is speculated to provide high-density nucleation sites, reduce the interfacial energy and then promote the mineralization process. The AKP-Ca complex could be incorporated into the nucleation sites and become a part of the mineralized nodules (Zhang et al, 2015;Jie et al, 2018). extracellular matrix (ECM) of osteoblasts could stimulate bone formation (Jie et al, 2018;Boskey et al, 2008).…”
Section: Attenuation Of H 2 O 2 -Suppressed Differentiation and Mineralization In Mc3t3-e1 Cellsmentioning
Phosphorylated peptide from Antarctic krill (P-AKP) was prepared by the dry-heating method with sodium pyrophosphate in order to improve its antioxidant activity and osteogenic activity. P-AKP exhibited more competitive DPPH• and OH• scavenging activities compared to the native Antarctic krill peptide (AKP). In hydrogen peroxide (H 2 O 2 )-induced oxidative damage of MC3T3-E1 cells, both AKP and P-AKP pretreatment could dose-dependently improve superoxide dismutase (SOD) and catalase (CAT) activities through attenuating the accumulation of reactive oxygen species (ROS) and malondialdehyde (MDA) production. Moreover, AKP and P-AKP prevented oxidative stress-induced down regulation of alkaline phosphatase (ALP) activity and matrix mineralization. Particularly, the promoting effects of P-AKP on the enzymatic antioxidant defense system, differentiation and mineralization was higher than that of AKP. These results suggested that phosphorylation might be a promising approach to improve the antioxidant and osteogenic activity of AKP, and P-AKP could be a beneficial agent for attenuating oxidative stress-related bone loss.
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