目的通过IL-1β体外诱导小鼠椎间盘软骨终板细胞建立退变细胞模型,并研究其作用机制。方法10 ng/ml IL-1β诱导小鼠椎间盘软骨终板细胞24h后,用cck-8法检时间点IL—1β对软骨终板细胞增殖作用的影响,透射电镜观察细胞退变情况,细胞免疫荧光和Western blot检测细胞退变相关蛋白表达。结果诱导组细胞较正常组细胞增殖减慢,细胞肥大化比例增加,出现线粒体肿胀、核扭曲、染色质边集,染色质及胞质松散等细胞坏死及凋亡表现,且co Ⅲ、Aggrecan表达下降,colX、MMP-1、MMP-3、MMP-13、TIMP-1表达增加。结论IL-1β可诱导软骨终板细胞发生退变。
We aimed to establish a disc degeneration cell model by IL-1β induction on mice enoplate chondrocytes, and analyze its mechanism. Firstly, we induced the endplate chondrocytes with 10 ng/ml IL-1β for 24 h. Then we used cck-8 to measure the proliferation during the different times. After that, we observed cell degeneration by TEM, and tested the expression of cell degeneration-related protein through immunofluorescence and Western blot. The results showed that the proliferation rate of induced cells was decreased compared with normal ones. The proportion of hypertrophy cell in induced group was higher too. Besides, there were many markers of meronecrobiosis and apoptosis in induced cells such as mitochondrial swelling, nucleus distortion, chromatin margination, chromatin and streaming loose. Moreover, we found the protein expression of col Ⅱ and aggrecan decreased, but col X, MMP-1, MMP-3, MMP-13, and TIMP-1 increased in induced group. We concluded that IL-1β can induce endplate chondrocytes to be degeneration.