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Abstract: FR-PO493

Specific Knockdown of WNT8b Expression Protects Against Phosphate-Induced Calcification in Vascular Smooth Muscle Cells by Inhibiting the Wnt/β-Catenin Signaling Pathway

Session Information

Category: Bone and Mineral Metabolism

  • 401 Bone and Mineral Metabolism: Basic

Authors

  • Yao, Li, The First Affiliated Hospital of China Medical University, Shenyang, China
  • Sheng, Zitong, The First Affiliated Hospital of China Medical University, Shenyang, China
  • Xu, Tianhua, The First Affiliated Hospital of China Medical University, Shenyang, China
  • Wan, Pengzhi, The First Affiliated Hospital of China Medical University, Shenyang, China
  • Binyao, Tian, The First Affiliated Hospital of China Medical University, Shenyang, China
Background

In the last 10 years, the prevalence, significance, and regulatory mechanisms of vascular calcification have gained increasing recognition. The aim of this work is to study the action of WNT8b on the disease development of phosphate-induced vascular calcification through its effect on vascular smooth muscle cells (VSMCs) in vitro by inactivating the Wnt/β-catenin signaling pathway.

Methods

In order to find out the effect of WNT8b on the Wnt/β-catenin signaling pathway and vascular calcification in vitro, β-glycerophosphate (GP) induced T/G HA-VSMCs were treated with siRNA against WNT8b, LiCl (Wnt agonists), and both, respectively. The mRNA and protein levels of WNT8b, α-SMA, calcification associated molecules, the Wnt signaling pathway related molecules were determined by reverse transcription quantitative polymerase chain reaction (RT-qPCR) and western blot analysis. TOP/FOP-Flash reporter assay was performed to detect transcription activity mediated by β-catenin.

Results

si-WNT8b reduced calcium deposition and activity of ALP, increased α-SMA level, and dropped BMP2, Pit1, MSX2, and Runx2 levels, while stimulation of LiCl worsened β-GP-induced calcium deposition, increased activity of ALP, reduced α-SMA expression level. si-WNT8b resulted in reductions in WNT8b, Fzd4, β-catenin, p-GSK-3β, and cyclin-D, whereas enhancement of p-β-catenin and GSK-3β , indicating si-WNT8b could alter the Wnt/β-catenin signaling pathway thus to hamper the vascular calcification in T/G HA-VSMC, which further demonstrated by TOP/FOP FLASH assay and detection of β-catenin expression level in the nucleus.

Conclusion

Taken together, we conclude that WNT8b knockdown abolishes phosphate-induced vascular calcification in VSMCs by inhibiting the Wnt/β-catenin signaling pathway.