压应力对人软骨终板细胞MMP-1、3、13和TIMP-1表达的影响研究

摘要

目的 观察压应力对体外培养人软骨终板细胞分泌基质金属蛋白酶-1、3、13 (MMP-1、MMP-3、MMP-13)和基质金属蛋白酶组织抑制因子1(TIMP-1)的影响.方法 将6个月意外流产胎儿软骨终板,进行软骨终板细胞的分离、培养.将第2代软骨细胞按实验要求种板培养,分别施加周期性压应力(压力大小0.8 MPa,频率为0.1Hz),分别连续加载2h、6h和12h.根据压应力作用时间分为2h组、6h组和12 h组,以空白组为对照,测定各组细胞上清液中MMP-1、MMP-3、MMP-13和TIMP-1的表达.结果 贴壁后的软骨终板细胞成短梭形、三角形以及多边形,呈典型的“铺路石”状.随着细胞加压时间延长,长梭形细胞数目明显增加,去分化现象明显,老化速度加快,坏死细胞明显增多.施加压应力第3天,6h组和12h组MMP-1、MMP-3、MMP-13表达明显增高,TIMP-1明显下降,且加压时间越长,改变越明显.24h组分别与各组相比较有统计学意义(P＜0.05).结论 长期压应力可使MMP-1、MMP-3、MMP-13表达增高,TIMP-1表达降低,并抑制细胞增殖,同时MMP-1、MMP-3、MMP-13表达水平与压应力作用时间相关.%Objective To investigate the effects of compressive stress on the secretion of matrix metalloproteinases-1,3,13 (MMP-1,MMP-3,MMP-13) and matrix metalloproteinase tissue inhibitor 1 (TIMP-1) in human cartilage endplate cells in vitro.Methods The cartilage endplate of 6 month aborted fetus was isolated and cultured.The second generation ofchondrocytes were cultured under the cyclic compressive stress (pressure size 0.8 MPa and frequency 0.1 Hz) with different duration (2 h,6 h and 12 h).Specimens were divided into group 2 h,group 6 h and group 12 h according to the duration of stress.The expression ofMMP-1,MMP-3,MMP-13 and TIMP-1 in the supematant of each group were detected.Results The endplate cells of the adherent wall appeared short fusiform,triangular and polygonal,typical of "cobblestone".With the increasing time of compressive stress,the spindle cell counts,the dedifferentiation phenomenon,the aging rate and the necrotic cell counts increased gradually.On the third day after stress,the expression of MMP-1,MMP-3 and MMP-13 were increased,while TIMP-1decreased in 6 h group and 12 h group.The longer the compression time,the more obvious the changes occurred in the 12 h group,these differences were statistically significant (P＜0.05) compared to other groups.Conclusion Long-term compressive stress increased the expression of MMP-1,MMP-3,MMP-13,decreased the expression of TIMP-1 and inhibited the proliferation of cells,and these changes were related to the duration of stress.