Calcium phosphate crystals promotes vascular calcification through BMP-2/Smad signaling
10.3760/cma.j.issn.1001-7097.2018.08.008
- VernacularTitle:磷酸钙晶体通过BMP-2/Smad信号通路促进血管钙化
- Author:
Yaorong LIU
1
;
Hao YAN
;
Zhenyuan LI
;
Li GONG
;
Xiaoxiao YANG
;
Liou CAO
;
Zhaohui NI
;
Jiaqi QIAN
;
Wei FANG
Author Information
1. 200127,上海交通大学医学院附属仁济医院肾脏科上海市腹膜透析研究中心分子细胞(肾病)实验室
- Keywords:
Renal insufficiency,chronic;
Calcinosis;
Calcium phosphates;
Myocytes,smooth muscle
- From:
Chinese Journal of Nephrology
2018;34(8):608-615
- CountryChina
- Language:Chinese
-
Abstract:
Objective To investigate the role of BMP-2/Smad signaling pathway in the osteogenic differentiation of human aortic smooth muscle cells (HASMCs) caused by hyperphosphatemia -induced calcium phosphate (CaP) crystals.Methods High-phosphate medium was incubated at 37℃ for 3 days.CaP crystals and supernatant were isolated by ultracentrifugation.Scanning electron microscope and energy dispersive X-ray spectroscopy were performed for analysis of physicochemical characteristics of CaP crystals.HASMCs were cultured in vitro,and divided into high-phosphate,control,crystals and supernatant groups.Calcification was visualized by Alizarin red staining.Calcium loads in cells were quantified by o-cresolphthalein complexone method.Protein expression of bone morphogenetic protein-2 (BMP-2),Runt-related transcription factor 2 (RUNX2),osteopontin (OPN),phospho-Smad1/5/9 (p-Smad1/5/9) were quantified by Western blotting.After knockdowns of BMP-2 and Smad1 with small hairpin RNA (shRNA) interfering respectively in HASMCs,protein expressions were measured by Western blotting.Results High-phosphate medium induced the formation of CaP crystals.Compared with the cells in control group,CaP crystals significantly induced HASMCs calcification,increased calcium loads and up-regulated the levels of BMP-2,RUNX2 and OPN proteins (all P ＜ 0.05).After the addition of CaP crystals into HASMCs,the level of p-Smad 1/5/9 protein peaked at 30 min (P ＜ 0.05).After BMP-2 was knocked down in HASMCs,the expression of p-Smad1 caused by CaP crystals was blocked completely,and the expressions of RUNX2 and OPN caused by CaP crystals were reduced significantly (all P ＜ 0.05).After Smad1 was knocked down in HASMCs,the expressions of RUNX2 and OPN caused by CaP crystals were decreased significantly (all P ＜ 0.05).Conclusions Hyperphosphatemia-induced CaP crystals promoted osteogenic differentiation of HASMCs through the BMP-2/Smad signaling pathway.
