【摘要】 目的探讨RNA干扰蛋白聚糖酶-1(aggrecanase-1)对大鼠肋软骨细胞基质代谢的影响。方法体外分离培养大鼠肋软骨细胞,采用脂质体转染试剂将针对aggrecanase-1的载体质粒转染软骨细胞,观察转染后细胞生长曲线、细胞形态的变化,RT-PCR检测aggrecanase-1mRNA水平的变化,Westernblotting检测蛋白多糖(aggrecan)的变化。结果RNA干扰aggreanase-1对细胞生长速度及形态无明显影响,可明显降低aggrecanase-1的mRNA表达水平(P<0.05),增加aggrecan的含量(P<0.05)。结论抑制aggrecanase-1可减少蛋白多糖的降解,RNAi是一种有效的研究软骨细胞基质代谢的工具。更多还原

【Abstract】 Objective To study the effect of RNA interference (RNAi)-mediated aggrecanase-1 gene silencing on extracellular matrix metabolism of cultured rat costochondral chondrocytes. Methods Rat costochondral chondrocyte monolayers were obtained by microdissection and digestion. The growth and morphological changes of the chondrocytes were observed after RNAi of aggrecanase-1 gene. The mRNA expression of aggrecanase-1 was detected by RT-PCR method, and aggrecan content was determined by Western blotting. Results The specific inhibition of aggrecanase-1 by RNAi produced no adverse effect on the morphology and growth of the chondrocytes. The mRNA of aggrecanase-1 decreased and aggrecan content increased significantly after transfection of the chondrocytes. Conclusion Inhibition of aggrecanase-1 decreases aggrecan degradation in cultured rat chondrocytes. RNAi technique can be a useful means for studying extracellular matrix metabolism in the cartilage.更多还原