MiR‐708‐5p/Pit‐1 axis mediates high phosphate‐induced calcification in vascular smooth muscle cells via Wnt8b/β‐catenin pathway

Abstract

Abstract Recently, the underlying mechanism of vascular calcification (VC) has been partially elucidated. However, it is still high incidence, and no effective treatment has been found. This study aims at figuring out the underlying mechanisms of microRNA‐708‐5p (miR‐708‐5p)/sodium‐phosphate transporter 1 (Pit‐1) axis in high phosphate (HP)‐induced VC of T/G HA‐VSMCs. Alizarin Red S staining was used to evaluate calcium salt deposition, and the activity of alkaline phosphatase (ALP) was determined by measuring the absorbance at 405 nm. RT‐qPCR and Western blot were performed to assess the levels of miR‐708‐5p and Pit‐1, the levels of ALP, Pit‐1, β‐catenin, glycogen synthesis kinase 3 β (GSK3β), and p‐GSK3β proteins, respectively. The interaction between miR‐708‐5p and Pit‐1 was validated by luciferase reporter assay. Our findings illustrated that miR‐708‐5p was downregulated and Pit‐1was upregulated in HP‐induced VC. MiR‐708‐5p mimics inhibited HP‐induced VC. Further experiments demonstrated that miR‐708‐5p targets Pit‐1. In addition, miR‐708‐5p inactivates the Wnt8b/β‐catenin pathway via targeting Pit‐1 to reduce HP‐induced VC. MiR‐708‐5p has a crucial effect on VC via targeting Pit‐1 and inhibiting Wnt8b/β‐catenin pathway, it may serve as a new target for VC treatment.

Keywords

• high phosphate
• MiR‐708‐5p
• pit‐1
• vascular calcification