Effect of bone morphogenetic protein-2 on human heart valve interstitial cells calcification in vitro
10.3760/cma.j.issn.0253-3758.2015.08.016
- VernacularTitle:骨形态发生蛋白2在人心脏瓣膜间质细胞体外钙化中的作用
- Author:
Qiuyang LING
1
;
Ting WU
;
Ting YE
;
Zhaoliang SU
;
Gangjun ZONG
Author Information
1. 214044,安徽医科大学无锡临床学院
- Keywords:
Stromal cells;
Heart valves;
Bone morphogenetic proteins
- From:
Chinese Journal of Cardiology
2015;43(8):724-727
- CountryChina
- Language:Chinese
-
Abstract:
Objective To establish human heart valve interstitial cells calcification culture model in vitro,and observe the effect of bone morphogenetic protein-2 (BMP-2) on calcification of human heart valve interstitial cells.Methods Human heart valve interstitial cells were cultured in vitro,and divided into control group:cells were cultured in conventional media plus recombinant human BMP-2 treatmnent and experimental group:besides above treaments,calcification inducers (recombinant hunan BMP-2,β-glycerophosphate,L-ascorbic acid,dexamethasone) were added to the culture media.The two group of cells were cultured for 14 days and were stained by Von Kossa,then the cell calcification was observed in this valvular interstitial cells calcification culture model in vitro.Protein expression of intercellular adhesion molecule 1 (ICAM-1),interleukin 8,BMP-2 and BMP-4 was determined by Western blot and BMP-2 secretion was measured by ELISA.Results In the control group,the structure of human heart valve interstitial cells was clear,and the spindle and radial growth shaped cellular morphology was visible,and Von Kossa staining was negative.In the experimental group,the nuclei become darker in color,and granular sediment distribution was seen surrounding cells,and Von Kossa staining was positive,the cells were forming nodules of calcification.The protein expression of ICAM-1,interleukin 8,BMP-2 and BMP-4 in the experimental was significantly higher than that of the control group (all P ＜ 0.05).The expression of BMP-2 in the experimental group was also significantly higher than that in control group ((92.5 ±4.9) pg/ml vs.(22.2 ± 1.9) pg/ml,P ＜ 0.05).Conclusion Human BMP-2,β-glycerophosphate,L-ascorbic acid,and dexamethasone can induce human heart valve interstitial cells calcification and enhance inflammation in vitro by stimulating the secretion of BMP-2.
