Open AccessThe intermediate‐conductance calcium‐activated potassium channel KCa3.1 contributes to alkalinization‐induced vascular calcification in vitro
Author(s) -
Bai Yaling,
Xu Jinsheng,
Yang Shuo,
Zhang Huiran,
He Lei,
Zhou Wei,
Cheng Meijuan,
Zhang Shenglei
Publication year - 2021
Publication title -
journal of clinical laboratory analysis
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.536
H-Index - 50
eISSN - 1098-2825
pISSN - 0887-8013
DOI - 10.1002/jcla.23854
Subject(s) - calcification , vascular smooth muscle , runx2 , alkaline phosphatase , osteopontin , western blot , chemistry , small hairpin rna , in vitro , calcium , microbiology and biotechnology , medicine , endocrinology , osteoblast , biology , biochemistry , gene knockdown , enzyme , apoptosis , smooth muscle , gene , organic chemistry
Objective In order to find new strategies for the prevention of vascular calcification in uremic individuals especially treated by dialysis and develop novel therapeutic targets in vascular calcification, we explore the role of KCa3.1 in alkalinization‐induced VSMCs calcification in vitro. Method Rat VSMCs calcification model was established by beta‐glycerophosphate (β‐GP, 10 mM) induction. The pH of Dulbecco's modified Eagle's medium (DMEM) was adjusted every 24 h with 10 mM HCl or 10 mM NaHCO 3 . The mineralization was measured by Alizarin Red staining and O‐cresolphthalein complex one method. mRNA and protein expression were detected by RT‐PCR and Western blot or immunofluorescence. Ca2+ influx was measured by Elisa. Result The results indicated that alkalization induced an increase in Ca2+ influx to enhance VSMCs calcification. Furthermore, the increase of calcification was associated with the expression of KCa3.1 via advanced expression of osteoblastic differentiation markers alkaline phosphatase (ALP) and Runt‐related transcription factor 2 (Runx2). Blocking KCa3.1 with TRAM‐34 or shRNA vector can significantly lowered the effects of calcification in the activity of ALP and Runx2 expression. Conclusion Together all, our studies suggested that alkalinization can promote vascular calcification by upregulating KCa3.1 channel and enhancing osteogenic/chondrogenic differentiation by upregulating Runx2. The specific inhibitor TRAM‐34 and KCa3.1‐shRNA ameliorated VSMCs calcification by downregulating KCa3.1.