CAJ | 학술논문

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。
배경：조성가체주위골용해적완정분자궤제목전상미완전청초。가체주위골용해、흡수시인공관절송동적전형병리생리과정。백세포개소1통과MAPK신호통로영향골흡수진정。 　　목적：실험통과관찰siRNA침묵백세포개소1수체상관격매4(interleukin-1 receptor-associated kinase-4， IRAK-4)기인표체대인성골양세포주MG63적MAPK신호전도통로적영향，위방치인공관절치환후가체주위골용해제공실험기출。 　　방법：이Lipofectamine 2000위재체장IRAK-4-siRNA전염입MG63세포。실험분위3조，공백조불가입임하전염시제；대조조이 scrambled siRNA 서렬진행전염；침묵조이특이성 IRAK-4-siRNA 서렬진행전염。Western blot검측파세포세포외조절단백격매、c-Jun안기말단격매화p38MAPK기인적표체。 　　결과여결론：여대조조상비，파기인침묵조적IRAK-4 mRNA급단백표체수평현저강저(P<0.05)。MG63세포 IRAK-4표체하조후，여공백조화대조조상비，c-Jun 안기말단격매1/2P46、린산화세포외조절단백격매1/2、린산화p38MAPK표체하조분별위62%，64%，68%(P <0.05)。결과증실，siRNA침묵IRAK-4기인억제인성골양세포주MG63세포외조절단백격매、c-Jun안기말단격매화p38MAPK기인적표체。
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-ERK1/2 and p-p38MAPK (P<0.05). IRAK-4 silencing inhibited ERK, JNK and p38MAPK expression in osteoblast-like cells.