PTH1-34 blocks advanced glycation end products′ negative osteogenesis to bone marrow mesenchymal stem cells through Wnt/β-catenin signaling pathway / 中华内分泌代谢杂志

ABSTRACT

Objective:To observe whether parathyroid hormone (PTH) 1-34 can block the negative osteogenesis of advanced glycation end products (AGEs) on bone marrow mesenchymal stem cells (BMSCs) in rats and its possible signaling pathways.Methods:BMSCs from 4-week-old SD rats were isolated and cultured with whole bone marrow method. The osteogenic induction fluid, bovine serum albumin(BSA), AGEs, AGEs combined with PTH1-34 were pretreated respectively. After 7 days, realtime fluorescence quantitative PCR (RT-PCR) was used to measure alkaline phosphatase (ALP), collagen-Ⅰ (COL-Ⅰ) mRNA expression level, and the expressions of β-catenin, osterix (OSX), runt-related transcription factor 2 (RUNX2), and receptor for advanced glycation end products protein were examined by Western blotting. Alizarin red staining mineralized nodules and quantitative detection were performed on 21 days. After the addition of Wnt pathway specific blocker Dickkopf-1 (DKK1) (1 μg/ml) to 10 -8 mmol/L of PTH1-34, the protein expression levels of β-catenin, OSX, and RUNX2 were detected again by Western blotting. Results:AGEs significantly inhibited the expression of ALP, COL-Ⅰ mRNA, β-catenin, OSX, and RUNX2 proteins ( P<0.05). PTH1-34 inhibited AGEs after pretreatment, the expression of ALP, COL-Ⅰ mRNA, and β-catenin, OSX, RUNX2 protein were higher than those of AGEs group ( P<0.05). The mineralized nodules stained with alizarin red showed reddish brown and increased OD value was detected quantitatively in PTH1-34 group, which was higher than that of AGEs group ( P<0.05). DKK1(1 μg/ml) reduced the expression of β-catenin, OSX proteins as compared with the BSA group ( P<0.05). Conclusion:PTH1-34 blocks AGEs′ negative osteogenesis to BMSCs through Wnt/β-catenin signaling pathway.