Aggrecanase-2表达沉默对体外培养的软骨细胞基质代谢的影响  

Effects of Inhibiting Aggrecanase-2 Gene Expression by RNA Interference on Cultured Chondrocyte Extracellular Matrix in Vitro

作　　者：王正辉[1] 杨壮群[2] 吴宝俊[1] 常会敏[1] Kamal Mustafa 卢晓云[4] 

机构地区：[1]西安交通大学第二附属医院耳鼻喉-头颈外科,陕西省西安市710004 [2]西安交通大学第一附属医院整形烧伤科,陕西省西安市710061 [3]挪威卑尔根大学牙科学院 [4]西安交通大学生命科学与技术学院,陕西省西安市710049

出　　处：《组织工程与重建外科杂志》2012年第5期245-248,共4页Journal of Tissue Engineering and Reconstructive Surgery

基　　金：国家自然科学基金(81000416);中央高校基本科研业务费专项资金(西安交通大学国际合作类;交叉学科类);西安交通大学第二附属医院人才培养基金[RC(XM)201102];西安交通大学第二附属医院重点项目基金[YJ(ZD)201103]

摘　　要：目的探讨RNA干扰蛋白聚糖酶-2(Aggrecanase-2)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-2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-2 mRNA水平的变化;Western Blot检测蛋白多糖(Aggrecan)的变化。结果RNA干扰Aggreanase-2对细胞生长速度及形态无明显影响,可明显降低Aggrecanase-2的mRNA表达水平(P<0.05),增加Aggrecan的含量(P<0.05)。结论抑制Aggrecanase-2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。Objective To evaluate the effect of RNAi mediated aggrecanase-2 on rat costochondral chondrocytes extra- eelluler matrix. Methods Rat costochondral chondrocytes were obtained by mierodissection and digestion, and cultured in monolayer. The growth velocity and morphological changes of chondrocytes were observed after transfection. The mRNA of aggrecanase-2 expression was detected by RT-PCR method and aggrecan content was detected by western blot. Results The specific inhibition of aggrecanase-2 by RNAi had no positive effect on the morphology and growth velocity of the chondrocytes. The express of mRNA of aggrecanase-2 was decreased significantly. RNAi significantly increased the aggreean content of chondrocytes treated. Conclusion Inhibition of aggrecanase-2 may decrease aggrecan degradation. RNAi technology can be a useful tool for studying degenerative processes in cartilage,

关 键 词：软骨细胞蛋 白聚糖酶-2 RNA干扰 

分 类 号：Q786[生物学—分子生物学]