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目的 观察风湿性心脏瓣膜间质细胞的生物学特征.方法 收集风湿性二尖瓣(22例)和正常二尖瓣(10例)标本,常规石蜡包埋后行苏木素-伊红(HE)和胶原纤维(VG)染色;新鲜标本分选瓣膜间质细胞分别行免疫细胞化学、Hoechst 33342染色和Western blot检测.结果 与正常瓣膜比较,风湿性二尖瓣表现为纤维化(22例)、钙化(20例).体外实验表明正常瓣膜间质细胞无特殊的生长形态,免疫表型为波纹蛋白[vimentin(+)]、α-平滑肌肌动蛋白[α-SMA(-)]、碱性磷酸酶[ALP(-)];风湿性瓣膜间质细胞易聚集生长成,免疫表型为vimentin(+)、α-SMA(+)、ALP(+).而且风湿性瓣膜间质细胞的Ⅰ型胶原纤维量(3倍)和凋亡细胞数(6倍)均明显高于正常瓣膜间质细胞(P＜0.05).结论 瓣膜间质细胞生物学特征的改变是导致瓣叶增厚、纤维化、钙化的主要因素.

Objective To identify biological characteristics of valvular interstitial cells (VICs) in rheumatic valvular disease. Methods Surgically excised rheumatic mitral valves ( n = 22) and normal mitral valves (n = 10) were collected and paraffin-embedded for Hematoxylin and Eosin (HE) and van gieson (VG) staining. Rheumatic and normal VICs were isolated from fresh mitral valves and analyzed using immunocytochemistry, Western blotting and Hoechst 33342 staining. Results Rheumatic mitral valves showed fibrosis (n = 22) and calcification (n = 20). The phenotype of normal VICs was vimentin ( + ),α-smooth muscle actin (a-SMA) ( - ) and alkaline phosphatase (ALP) ( - ), while rheumatic VICs showed vimentin ( + ), α-SMA ( + ) and ALP ( + ). The rheumatic VICs secreted abundant type Ⅰ collagen (3-fold) and contained more apoptotic cells (6-fold) than the normal VICs. Conclusion The change in biological characteristics of rheumatic VICs contributed to thickness, fibrosis and calcification seen in rheumatic valvular samples.