ALK2对小鼠髁突软骨细胞增殖分化的影响

Effects of ALK2 on the Proliferation and Differentiation of Condylar Chondrocytes in Mice

目的:探讨ALK2对小鼠髁突软骨细胞增殖及成软骨分化的作用。方法:采用酶消化法提取小鼠髁突软骨细胞并鉴定。体外采用siRNA沉默小鼠髁突软骨细胞中Alk2基因的表达,通过MTT法检测细胞增殖。通过实时定量RT-PCR方法检测软骨细胞标志基因的表达。结果:倒置显微镜观察体外培养的小鼠髁突软骨细胞形态。免疫组化染色显示COLⅠ强阳性,AGGRECAN弱阳性,COLⅡ弱阳性。Pellet培养后实时定量RT-PCR检测发现ColⅡ,ColⅩ和ColⅠ表达上调。证实所分离的细胞是髁突软骨细胞。MTT结果显示Alk2沉默后细胞数量增加,实时定量RT-PCR结果显示Alk2沉默后软骨细胞基因表达下调,并有显著统计学意义。结论:ALK2抑制小鼠髁突软骨细胞的增殖,促进小鼠髁突软骨细胞的成软骨分化。

Objective：To study the effect of ALK2 on the proliferation and differentiation of mice condylar chondrocytes.Methods：Condylar chondrocytes（MCC）were obtained by enzyme digestion.Immunocytochemistry and real-time PCR were used to identify the isolated cells.SiRNA was used to silence the expression of Alk2 gene in MCC in vitro.Cell proliferation was determined by MTT.The expression of cartilage cell marker genes was analyzed by real-time PCR.Results：Immunocytochemical staining showed strong positive expression of COL I and AGGRECAN,and weak positive expression of COL II.Real-time PCR results showed that the expressions of cartilage related genes such as COL I,COL II,COL X were increased.After silencing ALK2,MTT results showed that cell number was increased and the expression of cartilage related genes was significantly down-regulated.Conclusion：ALK2could inhibit the proliferation of MCC and promote the chondrogenic differentiation of MCC.