Vascular smooth muscle cells transformation induced by high phosphate enviroment in vitro
10.3760/cma.j.issn.1001-7097.2009.02.010
- VernacularTitle:体外高磷环境下血管平滑肌细胞转分化过程的研究
- Author:
Ningning WANG
;
Xiaoyun WANG
;
Junwei YANG
;
Changying XING
;
Xiaobing JU
;
Bin SUN
;
Shuqin ZHOU
;
Ping WEN
;
Mingxia XIONG
- Publication Type:Journal Article
- Keywords:
Myocytes,smooth muscle;
Caleinosis;
Muscle,smooth,vascular;
Phosphorus
- From:
Chinese Journal of Nephrology
2009;25(2):116-121
- CountryChina
- Language:Chinese
-
Abstract:
Objective To observe the steps of vascular smooth muscle cells (VSMCs) calcification induced by high phosphate enviroment in vitro. Methods VSMCs were incubated with high phosphate (2.5 mmol/L or 3.5 mmool/L) medium for different times. Expression of core binding factor α1(Cbfα1), osteopontin(OP), collagen type Ⅰ(Col Ⅰ), osteocalcin(OC) and α-smooth muscle actin (α-SMA) was investigated by Western blot, immunofluorcscencc staining and real time PCR. Mineral deposition was assessed by von Kossa aad Alizarin red staining. Ultrastructure of VSMCs calcification was observed by electron microscopy (EM). Results Up-regulated expression of osteoblast-specific transcription factor Cbfα1 in the nuclei oceured at as early as 12 hours. The protein of Col Ⅰ and OP was up-regalated when VSMCs were incubated in high phosphate medium for 3 days, and content of OC increased at the time of 6 days. When cultured in 2.5 mmol/L phosphate medium for 15 days, VSMCs lost their lineage marker α-SMA, developed granular calcium deposits. Moreover, the results of real time PCR indicated mRNA level of OP and Col Ⅰ increased at day 1, OC increased at day 5 and α-SMA level decreased at day 10, respectively. Ultrastructural analysis also confirmed the presence of collagen and matrix vesicles in the cells. Conclusion VSMCs phenotype transformation induced by high phosphate enviroment is an orchestrated, highly regulated process.
