IL-17对破骨细胞中MMP-9表达水平的影响及其意义

Effects of IL-17 on expressions of MMP-9 in osteoclasts and their significances

目的:探讨白细胞介素17(IL-17)对破骨细胞中基质金属蛋白酶9(MMP-9)表达水平的影响,为研究IL-17在正畸力致牙根吸收过程中的作用提供依据。方法:培养小鼠单核/巨噬系细胞RAW264.7,将细胞分为实验组和对照组,实验组细胞培养液中加入梯度浓度0.1、1.0、10.0和100.0μg·L-1IL-17,作为0.1、1.0、10.0和100.0μg·L-1 IL-17组,对照组细胞培养液中不加入IL-17。对各组RAW264.7细胞进行破骨细胞诱导,诱导第5天时采用抗酒石酸酸性磷酸酶(TRAP)染色鉴定破骨细胞诱导成功。采用酶联免疫吸附(ELISA)法检测破骨细胞诱导第5天时各组RAW264.7细胞培养液中MMP-9表达水平;采用逆转录-聚合酶链式反应(RT-PCR)法检测破骨细胞诱导第5天时各组RAW264.7细胞培养液中MMP-9 mRNA表达水平。结果:TRAP染色,RAW264.7细胞诱导第5天时可见TRAP染色阳性细胞,体积大,多核(细胞核≥3个)。ELISA和PT-PCR法检测,经破骨细胞诱导第5天时,与对照组比较,1.0、10.0和100.0μg·L-1IL-17组RAW264.7细胞培养液中MMP-9和MMP-9mRNA表达水平升高(P<0.05);与1.0μg·L-1 IL-17组比较;10.0和100.0μg·L-1IL-17组RAW264.7细胞培养液中MMP-9和MMP-9mRNA表达水平升高(P<0.05);与10.0μg·L-1IL-17组比较,100μg·L-1IL-17组RAW264.7细胞培养液中MMP-9和MMP-9mRNA表达水平升高(P<0.05)。结论:IL-17可以促进破骨细胞中MMP-9表达,且其促进作用随着IL-17浓度的升高而增强。

Objective：To explore the effects of interleukin-17（IL-17）on the expression levels of matrix metalloproteinase-9（MMP-9）in the osteoclasts,and to provide the references for the study on the effect of IL-17 on the orthodontic force-induced root resorption.Methods：The mouse monocytes/macrophages RAW264.7 were cultured and divided into experiment group and control group;the culture medium in experiment group were added with the gradient concentrations of 0.1,1.0,10.0,and 100.0μg·L-1 IL-17,and set as 0.1,1.0,10.0,and100.0μg·L-1 IL-17 groups,and the culture medium in control group was IL-17-free.The RAW264.7cells in various groups were induced to form osteoclasts and identified by TRAP staining on the 5th day after induction.ELISA method was used to detect the expression levels of MMP-9in the culture medium of RAW264.7cells in various groups when the osteoclasts were induced for 5d;RT-PCR method was used to detect the expression levels of MMP-9mRNA in the culture medium of RAW264.7cells in various groups when the osteoclasts were induced for5 d.Results：The TRAP staining results showed TRAP staining positive cells were found,with large volume and large number of nucleus（nucleus≥3）on the 5th day after induction.The ELISA and RT-PCR results showed the expression levels of MMP-9and MMP-9 mRNA in the culture medium of RAW264.7cells in 1.0,10.0and100.0μg·L-1 IL-17 groups were increased compared with control group（P〈0.05）;the expression levels of MMP-9and MMP-9mRNA in the culture medium of RAW264.7cells in 10.0and 100.0μg·L-1 IL-17 groups were increased compared with 1.0μg·L-1 IL-17group（P〈0.05）;the expression levels of MMP-9and MMP-9mRNA in the culture medium of RAW264.7cells in 100.0μg·L-1 IL-17 group were increased compared with10.0μg·L-1 IL-17 group（P〈0.05）.Conclusion：IL-17 can promot the expressions of MMP-9 in the osteoclasts,and the promoting effect is enhanced with the increasing of IL-17 concentration.