Inhibitory effect of interleukin-1 receptor-associated kinase-4 silencing on mitogen-activated protein kinases expression in human osteoblast-like cells / 中国组织工程研究

ABSTRACT

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.