IL-17A对成骨细胞系MC3T3-E1细胞中明胶酶表达的影响及其作用机制

目的 检测IL-17A对MC3T3-E1细胞中明胶酶表达的影响,并探讨其作用机制。方法 采用小鼠重组细胞因子IL-17A作用于MC3T3-E1细胞,通过Real-time PCR和ELISA分别检测MMP-2及MMP-9在mRNA水平及蛋白水平上的表达情况;通过免疫荧光和Western Blot检测NF-κB的磷酸化水平的变化,并通过使用NF-κB阻断剂检测其在MMP-9表达中的影响。结果 IL-17A在mRNA水平及蛋白水平上均上调MMP-9的表达(P＜0.01),然而对MMP-2的表达无明显影响;IL-17A促进转录因子NF-κB的磷酸化水平(P＜0.01);阻断NFκB的活性可减弱IL-17A对MMP-9的上调作用(P＜0.05);IL-17A对MC3T3-E1细胞中TIMP-1及TIMP-2的表达没有影响。结论 IL-17A可以通过激活NF-κB促进MC3T3-E1细胞分泌MMP-9。

Effect of IL-17A on the expression of gelatinase in osteoblasts MC3T3-E1 cells and its mechanism

Objective To investigate the effect of IL-17A on gelatinase expression in MC3T3-E1 cells and to explore its mechanism.Methods MC3T3-E1 cells were stimulated by IL-17A. The expression of MMP-2 and MMP-9 at mRNA and protein levels were detected by Real-time PCR and ELISA respectively. Immunofluorescence and Western Blot were used to detect the IL-17A-induced activity of NF-κB. Chemical inhibitor of NF-κB was used to detect the involvement of NF-κB in IL-17A-induced MMP-9 expression in MC3T3-E1 cells. Results IL-17A upregulated MMP-9 expression at both mRNA and protein levels (P＜0.01), but had no effect on MMP-2 expression; IL-17A promoted phosphorylation of NF-κB (P＜0.01); inhibition of NF-κB attenuated IL-17A-induced MMP-9 expression(P＜0.05). IL-17A did not affect the TIMP-1 and TIMP-2 expression at mRNA level.Conclusion IL-17A can promote the secretion of MMP-9 by MC3T3-E1 cells by activating NF-κB.