α-SMA的表达与软骨细胞的去分化  

Alpha-SMA expression and chondrocyte dedifferentiation

作　　者：季煜华[1] 曾耀英[1] 肇静娴[1] 

机构地区：[1]暨南大学组织移植与免疫研究中心,广东广州510632

出　　处：《第四军医大学学报》2007年第7期585-588,共4页Journal of the Fourth Military Medical University

基　　金：国家重点基础研究发展规划项目(G1999054303);国家自然科学基金面上项目(30600587);暨南大学引进人才启动基金项目(51205066)

摘　　要：目的:探讨细胞骨架蛋白α-SMA的表达与软骨细胞去分化之间的联系.方法:分离、培养大鼠肋生长板软骨细胞(RGCs),对原代至第5代RGCs进行细胞生长动力学分析,阿尔新兰染色、免疫细胞化学法和蛋白质印迹分别检测蛋白多糖,α-SMA和Ⅱ型胶原的表达.结果:随着传代次数的增加,前3代RGCs的每日倍增指数逐渐增加,随后不断下降.原代RGCs的阿尔新兰和Ⅱ型胶原染色均呈强阳性,仅个别细胞α-SMA弱阳性染色.随着传代次数的增加,阿尔新兰和Ⅱ型胶原阳性染色的细胞比例迅速下降,第3代以后的RGCs阿尔新兰染色均为阴性,第5代RGCs也仅有圆形细胞仍保持了Ⅱ型胶原阳性染色,而α-SMA阳性染色细胞的比例则不断增加.蛋白质印迹所揭示的Ⅱ型胶原和α-SMA蛋白表达的变化趋势与免疫细胞化学染色的结果一致.结论:α-SMA从无到有并随着RGCs去分化程度的增强而增加的表达模式,提示它是引起软骨细胞“去分化”的重要原因.AIM： To investigate the relationship between cytoskeleton α-SMA （alpha-smooth muscle actin） expression and dedifferentiation of chondrocyte. METHODS ： Rat costochondral growth plate chondrocytes （RGCs） were isolated and cultured. Growth kinetics of RGCs during serial passage was analyzed, and α-SMA, collagen type Ⅱ and GAG expressions of primary to 5th passage RGCs were detected by immunocytochemistry, Western Blot and Alcian blue staining, respectively. RESULTS： With increasing passage number, the index of duplication/day of RGCs rose during the first 3 passages and decreased afterwards. Primary RGCs expressed collagen type Ⅱ and were stained strongly for Aleian blue, and very few RGCs were positive for α-SMA. The percentage of collagen type Ⅱ and Alcian blue positive stained RGCs decreased with increasing passage number. Only the roundish cells of the 5th passage retained positive for collagen type Ⅱ , and Alcian blue staining of 3rd passage was negative, while α-SMA positive stained RGCs increased gradually. Western Blot results were consistent with immunocytochemical outcomes. CONCLUSION： As the degrees of RGCs dedifferentiation rise, α-SMA expression increases gradually. The expression pattern of α-SMA suggests that it might be an important factor in inducing chondrocyte dedifferentiation.

关 键 词：生长板 软骨细胞 去分化 Α-平滑肌肌动蛋白 

分 类 号：R363[医药卫生—病理学]