摘要
目的研究白细胞介素17(IL-17)是否促进主动脉瓣膜钙化形成及其可能机制。方法利用体外主动脉瓣膜间质细胞培养技术,瓣膜间质细胞培养传代3-5次后,采用简单随机抽样法分为两组:(1)对照组,细胞培养基中添加2 ml RPMI-1640完全培养液;(2)实验组,细胞培养基中添加2 ml RPMI-1640完全培养液和IL-17(50 ng/ml)。继续孵育48 h后,用细胞茜素红钙染色检测两组瓣膜间质细胞钙化情况,用Western blot和RT-PCR检测两组瓣膜间质细胞碱性磷酸酶(ALP)及骨形态蛋白2(BMP-2)的表达情况。结果实验组细胞中明显钙结节形成,对照组细胞中少量钙结节形成;与对照组比较,实验组细胞ALP和BMP-2蛋白(0.741±0.063比0.184±0.032;0.900±0.060比0.396±0.030,均为P〈0.01)与基因(0.236±0.004比0.106±0.003;0.523±0.052比0.194±0.047,均为P〈0.01)表达量均明显升高。结论 IL-17可促进主动脉瓣膜间质细胞向成骨样细胞转化。
Objective To investigate whether IL-17 could stimulate aortic valve interstitial cells transformation into osteoblast-like cells. Methods Human aortic valve interstitial cells were cultured via culture technology in vitro,and randomly divided into two groups after the subcultivation for three to five generations: the control group,adding 2 ml RPMI-1640; the experiment group,adding 2 ml RPMI-1640 and IL-17( 50 ng /ml). After 48 hours,the calcification of valve interstitial cells was detected by cell alizarin red staining and the expression of alkali phosphatase( ALP) and bone morphogenetic protein-2( BMP-2)was examined by RT-PCR and Western Blot analysis. Results Compared with the control group,valve interstitial cells in the experiment group had more calcium nodus and expressed more protein and mRNA of ALP( 0. 741 ± 0. 063 vs. 0. 184 ± 0. 032; 0. 900 ± 0. 060 vs. 0. 396 ± 0. 030,both P〈0. 01) and BMP-2( 0.236 ±0.004 vs. 0.106 ± 0.003; 0.523 ± 0.052 vs. 0.194 ± 0.047,both P〈0.01). Conclusions IL-17 can stimulate aortic valve interstitial cells transformation into osteoblast-like cell.
出处
《中国心血管杂志》
2014年第5期364-367,共4页
Chinese Journal of Cardiovascular Medicine
基金
国家自然科学基金项目(87210297)~~