Lithium chloride regulates odontogenic differentiation of dental pulp stem cells through phosphatidylinositol 3-kinase/Akt signaling pathways
Objective To explore the effect of Wnt/β-catenin pathway activator lithium chloride on the differentiation ability of dental pulp stem cells (DPSCs) and its related molecular mechanism.Methods The formation of mineralized nodules in DPSCs cultured in vitro was evaluated by alizarin red staining after 1 and 2 weeks of stimulation with different concentrations of lithium chloride (1 and 10 mmol/L).Real-time fluo-rescent quantitative reverse transcription PCR (RT-qPCR) was used to detect the effects of 1 mmol/L lithium chloride on the mRNA expressions of dentin sialophosphoprotein (DSPP),dentin matrix protein 1 (DMP1), alkaline phosphatase (ALP),bone sialoprotein (BSP) and osteocalcin (OCN) in DPSCs.LY192004 was used to inhibit the phosphoinositide-3-kinase/protein kinase B (PI3K/Akt) signaling pathway,and its effects on the formation of mineralized nodules and mRNA expression of mineralized related proteins in DPSCs were verified by alizarin red staining and RT-qPCR.Further,Western blot analysis was performed to analyze the phospho-rylation expression of Akt in DPSCs stimulated by 1 mmol/L lithium chloride with or without LY294002.Re-sults Compared with 0 mmol/L lithium chloride,1 mmol/L lithium chloride significantly promoted the for-mation of mineralization nodules and the mRNA expressions of DSPP,DMP1,BSP and ALP in DPSCs (P<0.05),while 10 mmol/L lithium chloride significantly inhibited the formation of mineralization nodules in DP-SCS (P<0.05).After adding 1 mmol/L lithium chloride and LY294002 (25 μmol/L) to culture DPSCs for 2 weeks,the increased formation of mineralized nodules and the expressions of DSPP,DMP1 and ALP mRNA induced by 1 mmol/L lithium chloride stimulation were inhibited,and the differences were statistically signifi-cant (P<0.05).Western blot results showed that lithium chloride (1 mmol/L) significantly promoted the phosphorylation of Akt in a time-dependent manner (P<0.05),while LY294002 significantly inhibited the phosphorylation of Akt (P<0.05).Conclusion Lithium chloride can regulate the odontogenic differentiation of DPSCs by regulating the PI3K/Akt signaling pathway.
lithium chloridedental pulp stem cellsodontoblastic differentiationphosphoinositide-3-kinaseprotein kinase B
万方数据
维普
