Phosphocreatine Promotes Osteoblastic Activities in H2O2-Induced MC3T3-E1 Cells by Regulating SIRT1/FOXO1/PGC-1α Signaling Pathway

Curr Pharm Biotechnol. 2021;22(5):609-621. doi: 10.2174/1389201021999201116160247.

Abstract

Background: Osteoporosis, characterized by bone loss, usually occurs with the increased bone resorption and decreased bone formation. H2O2-induced MC3T3-E1 cells are commonly used for the study of osteoblastic activities, which play a crucial role in bone formation.

Objective: This study aimed to investigate the effects of Phosphocreatine (PCr) on the osteoblastic activities in H2O2-induced MC3T3-E1 cells and elaborate on the possible molecular mechanism.

Methods: The Osteoprotegerin (OPG)/Receptor Activator of NF-κB Ligand (RANKL) ratio and osteogenic markers were detected to investigate the effects of PCr on osteoblastic activities, and the osteoblastic apoptosis was detected using Hochest staining. Moreover, oxidative stress, Adenosine Triphosphate (ATP) generation and the expression of Sirtuin 1 (SIRT1), Forkhead Box O 1 (FOXO1) and Peroxisome Proliferator-Activated Receptor Γ Coactivator-1α (PGC-1α) were also examined to uncover the possible molecular mechanism in H2O2-induced MC3T3-E1 cells.

Result: The results showed that PCr promoted the osteoblastic differentiation by increasing the expression levels of osteogenic markers of Alkaline Phosphatase (ALP) and Runt-related transcription factor 2 (Runx2), as well as increased the OPG/RANKL ratio and suppressed the osteoblastic apoptosis in H2O2-induced MC3T3-E1 cells. Moreover, treatment with PCr suppressed reactive oxygen species (ROS) over-generation and promoted the ATP production as well as increased the PGC-1α, FOXO1 and SIRT1 protein expression levels in H2O2-induced MC3T3-E1 cells.

Conclusion: PCr treatment could promote osteoblastic activities via suppressing oxidative stress and increasing the ATP generation in H2O2-induced MC3T3-E1 cells. In addition, the positive effects of PCr on osteoblasts might be regulated by SIRT1/FOXO1/ PGC-1α signaling pathway.

Keywords: FOXO1; PGC-1α.; Phosphocreatine; SIRT1; hydrogen peroxide; osteoblastic activities.

MeSH terms

  • 3T3 Cells
  • Alkaline Phosphatase / biosynthesis
  • Alkaline Phosphatase / drug effects
  • Animals
  • Apoptosis / drug effects
  • Core Binding Factor Alpha 1 Subunit / biosynthesis
  • Core Binding Factor Alpha 1 Subunit / drug effects
  • Forkhead Box Protein O1 / drug effects*
  • Hydrogen Peroxide / pharmacology*
  • Mice
  • Osteoblasts / drug effects*
  • Osteogenesis / drug effects*
  • Osteoprotegerin / drug effects
  • Osteoprotegerin / metabolism
  • Oxidative Stress
  • Peroxisome Proliferator-Activated Receptor Gamma Coactivator 1-alpha / drug effects*
  • Phosphocreatine / pharmacology*
  • RANK Ligand / drug effects
  • RANK Ligand / metabolism
  • Reactive Oxygen Species
  • Signal Transduction / drug effects*
  • Sirtuin 1 / drug effects*

Substances

  • Core Binding Factor Alpha 1 Subunit
  • Forkhead Box Protein O1
  • Foxo1 protein, mouse
  • Osteoprotegerin
  • Peroxisome Proliferator-Activated Receptor Gamma Coactivator 1-alpha
  • Ppargc1a protein, mouse
  • RANK Ligand
  • Reactive Oxygen Species
  • Runx2 protein, mouse
  • Tnfrsf11b protein, mouse
  • Tnfsf11 protein, mouse
  • Phosphocreatine
  • Hydrogen Peroxide
  • Alkaline Phosphatase
  • Sirt1 protein, mouse
  • Sirtuin 1