周期性静水压对人软骨细胞IL-1β和MMP-3代谢的影响

Effects of Periodic Hydrostatic Pressure on IL-1β and MMP-3 Metabolism of Human Chondrocytes

目的:以正常成人软骨细胞及OA软骨细胞为研究对象,探讨在静水压作用下正常软骨细胞IL-1β和MMP-3代谢的变化,以及与OA软骨细胞之间的差异。同时观察在力学信号刺激下软骨细胞的IL-1β和MMP-3代谢有无相关性。方法:将体外培养的正常成人膝关节软骨第3代软骨细胞随机分为加压组和对照组。甲苯胺蓝及Ⅱ型胶原免疫细胞化学染色进行细胞鉴定。将加压组细胞放入高压恒温静水压加压系统,冲入含有95%的空气和5%的CO2混合气体,以10MPa进行加压2h,共加压5d。分别于加压前、加压第3天、加压第5天后取加压组、对照组和OA组细胞培养液检测IL-1β、MMP-3含量。同时采用MTT法分析3组细胞的增殖情况。对三组细胞中IL-1β与MMP-3的水平做双变量相关分析,采用方差分析对三组标本间的IL-1β、MMP-3含量进行比较。结果:各组细胞IL-1β与MMP-3两者间存在正相关关系。OA组细胞中IL-1β与MMP-3含量高于加压组和对照组,加压组高于对照组。加压组、OA组软骨细胞的生长曲线与对照组相比增殖高峰降低,平台时间缩短。结论:10MPa间歇性静水压加压可抑制软骨细胞增殖,增加IL-1β与MMP-3分泌。

Objective： To investigate the changes oflL-1 β and MMP-3 metabolism in normal chondrocytes under hydrostatic pressure, and the difference between OA and loaded chondrocytes, with the normal adult and OA chondrocytes as the object. At the same time, to investigate the correlation between IL-1 and MMP-3 of chondrocytes in the mechanical stimuli. Methods： The third generation of the chondrocytes, which come from the knee articular cartilage of adult normal cultured in vitro, were randomly divided into loaded groups and control groups. Totuidine blue staining and type II collagen immunocytochemical staining were applied to identify the chondrocytes. The loaded group was put into the high hydrostatic pressure loading system filled with the mixed gas with 95% air and 5% CO2, periodically pressurized to 10 MPa pressure, 2h per day for 5days. The cell culture fluid of the loaded group, control group and OA group was used to detect the content oflL-1 and MMP-3 respectively in before compression, 3days and 5 days after compression. MTT assay analyzed the information of chondrocytes cell multiplication in three groups. The bivariate correlation was used to analysis the IL-1 and MMP-3 level. ANOVA was used to compare the IL-1 and MMP-3 content in three groups. Results： There was a positive correlation between IL-1 β and MMP-3. The content oflL-1 β nd MMP-3 in OA group was higher than that in the loaded and control group, while which in loaded group was higher than that in the control group. Compared with that in the control group, the proliferate peak decreased and the platform time shortened on the growth carve of the loaded and OA group. Conclusion： 10MPa intermittent hydrostatic pressure could inhibit the proliferation of chondrocytes, and increase the secretion of MMP-3 and IL-1 β.