白细胞介素1受体相关激酶4基因沉默抑制人成骨样细胞相关基因的表达  

Inhibitory effect of interleukin-1 receptor-associated kinase-4 silencing on mitogen-activated protein kinases expression in human osteoblast-like cells

作　　者：杨子波[1] 黄保丁[2] 向珊珊[3] 邬培慧[1] 张志奇[1] 廖威明[1] 

机构地区：[1]中山大学附属第一医院关节外科,广东省广州市510080 [2]中山大学附属第六医院骨科,广东省广州市510655 [3]江门市中心医院,广东省江门市529030

出　　处：《中国组织工程研究》2013年第46期7988-7993,共6页Chinese Journal of Tissue Engineering Research

基　　金：国家自然科学基金项目(81171709)~~

摘　　要：背景:造成假体周围骨溶解的完整分子机制目前尚未完全清楚。假体周围骨溶解、吸收是人工关节松动的典型病理生理过程。白细胞介素1通过MAPK信号通路影响骨吸收进程。目的:实验通过观察siRNA沉默白细胞介素1受体相关激酶4(interleukin-1 receptor-associated kinase-4,IRAK-4)基因表达对人成骨样细胞株MG63的MAPK信号转导通路的影响,为防治人工关节置换后假体周围骨溶解提供实验基础。方法:以Lipofectamine 2000为载体将IRAK-4-siRNA转染入MG63细胞。实验分为3组,空白组不加入任何转染试剂;对照组以scrambled siRNA序列进行转染;沉默组以特异性IRAK-4-siRNA序列进行转染。Western blot检测靶细胞细胞外调节蛋白激酶、c-Jun氨基末端激酶和p38MAPK基因的表达。结果与结论:与对照组相比,靶基因沉默组的IRAK-4 mRNA及蛋白表达水平显著降低(P<0.05)。MG63细胞IRAK-4表达下调后,与空白组和对照组相比,c-Jun氨基末端激酶1/2P46、磷酸化细胞外调节蛋白激酶1/2、磷酸化p38MAPK表达下调分别为62%,64%,68%(P<0.05)。结果证实,siRNA沉默IRAK-4基因抑制人成骨样细胞株MG63细胞外调节蛋白激酶、c-Jun氨基末端激酶和p38MAPK基因的表达。BACKGROUND： The molecular mechanism of periprosthesis osteolysis is not yet completely clear. Periprosthetic osteolysis and absorption is the pathological and physiological process typical of artificial joint loosening. Interleukin-1 can affect bone resorption process through a mitogen-activated protein kinases （MAPK） signaling pathway. OBJECTIVE： To explore the effects of siRNA-induced interleukin-1 receptor-associated kinase-4 gene （IRAK-4） silence on MAPK expression in MG63 cells, which maY provide experimental basis for treatment and prevention of Periprosthesis osteolysis. METHODS： The siRNA sequences of the target gene, IRAK-4, were constructed and transferred into MG63 cells using Lipofectamine 2000. There were three groups： blank group=MG63 cells, control group = MG63 cells transfected with scrambled IRAK-4siRNA, and silence group=MG63 cells transfected with specific IRAK-4 siRNA： The protein level Of extracellular regulated kinase （ERK）, c-Jun N-terminal kinase （JNK） and p38 mitogen-activated protein kinases （p38MAPK） were detected by western blot assay. RESULTS AND CONCLUSION： The expression of IRAK-4 mRNA and protein in the silence group was significantly decreased compared with the control group. Compared with the.blank and control groups, 48 hours after the transfection IRAK-4 gene silencing in MG63 cells decreased protein expression of p-JNK1/2P46, p-ERKI/2 and p-p38MAPK （P 〈 0.05）. IRAK-4 silencing inhibited ERK, JNK and p38MAPK expression in osteoblast-like cells.

关 键 词：组织构建 骨组织构建 假体周围骨溶解 无菌性松动 人成骨样细胞 白细胞介素1受体相关激酶4 RNA干扰 基因沉默 基因转染 促分裂素原活化蛋白激酶 国家自然科学基金 

分 类 号：R318[医药卫生—生物医学工程]