基质金属蛋白酶-9在大鼠终板软骨细胞中的表达及意义

Expression change of matrix metalloproteinase-9in the rat endplate chondrocytes and its significance

目的观察大鼠终板软骨细胞体外自然退变过程中基质金属蛋白酶(MMP)-9表达变化及意义。方法酶消化法及自然传代法获取大鼠腰椎终板软骨细胞,选取P1代及P7代并分别定义为实验组和对照组,倒置相差显微镜、HE染色、甲苯胺蓝染色及实时逆转录-聚合酶链反应(RT-PCR)鉴定终板软骨细胞。实时RT-PCR及Western免疫印迹检测MMP-9变化。结果实验组终板软骨细胞增殖速度减慢并呈梭型,Ⅱ型胶原、聚集蛋白聚糖表达均明显下调,MMP-9表达明显增加。结论 MMP-9可能参与终板软骨细胞退变过程,早期调节其表达有可能会阻止终板软骨细胞退变,为椎间盘退变防治提供新的理论依据。

Objective To study the expression change of matrix metalloproteinase-9 （MMP-9） and its significance in vitro model of natural degeneration of rat endplate chondrocytes. Methods The rat lumbar endplate chondroeytes were isolated by enzyme digestion and cultured. The one and seven generation cells were collected, and divided into experimental group （seven- generation） and control group （one-generation）. The cells were identified with inverted phase contrast microscope, HE staining, toluidine blue staining and real-time reverse transcription polymerase chain reaction （RT-PCR）. The expression of MMP-9 of cells were determined by reabtime RT-PCR and Western blot. Results The experimental group endplate chondrocytes were proliferative speed decreased and fusiform, the expression level of type lI collagen and aggrecan were significantly decreased. Furthermore, MMP-9 was significantly increased. Conclusions MMP-9 may be involved in the process of endplate chondrocytes degeneration. These findings implicate that prevention of degeneration of endplate chondrocytes can be achieved through regulating the expression of MMP9.