钙化性主动脉瓣疾病瓣膜间质细胞的生物学特征分析

Biological characteristics of valvular interstitial cells in calcific aortic valve disease

目的初步分析钙化性主动脉瓣疾病(calcific aortic valve disease,CAVD)瓣膜间质细胞的生物学特征,为后续研究奠定基础。方法采用组织块接种和免疫磁珠法分选正常主动脉瓣和CAVD瓣膜间质细胞,观察其形态学和行为学特征。通过免疫细胞化学染色和流式细胞术分析CAVD瓣膜间质细胞免疫表型的改变。结果与正常主动脉瓣瓣膜间质细胞相比,CAVD瓣膜间质细胞呈肌纤维母细胞和成骨细胞形态,当细胞达到一定密度后细胞自发性收缩、聚集样生长并形成钙化结节。体外培养的CAVD瓣膜间质细胞表达肌纤维母细胞标志物α-SMA和成骨细胞标志物碱性磷酸酶(ALP)。结论 CAVD瓣膜间质细胞生物学特征的改变可能是导致主动脉瓣增厚、钙化和交界融合的重要因素。

Objective To observe the biological characteristics of valvular interstitial cells in calcific aortic valve disease （CAVD）, so as to lay a foundation for future study. Methods Tissue culture inoculation and immunomagnetic bead separation method were used to isolate the valvular interstitial cells from the normal aortic valves and CAVD valves. The morphological and behavioral characteristics of the isolated valvular interstitial cells were observed. Immunocytochemistry and flow cytometry analysis were employed to determine cellular imrnunophenotype. Results Compared to normal valvular interstitial cells, CAVD valvular interstitial cells displayed a myofibroblast- and osteoblast like morphology. When the cell density reached a certain level, they spontaneously retracted from the neighboring areas and grouped into aggregates, forming calcific nodules. Furthermore, CAVD valvular interstitial cells cultured in vitro were positive for myofibroblast marker α-SMA and osteoblast marker alkaline phosphatase. Conclusion Biological characteristic change of CAVD valvular interstitial cells might be the major reason for the thickening, calcification, and commissural fusion in CAVD valvular samples.