骨桥蛋白对人膝骨关节炎软骨细胞透明质酸表达的影响

Effect of osteopontin on the expression of hyaluronic acid in human knee osteoarthritic chondrocytes

背景:在骨关节炎病程中,骨桥蛋白水平和透明质酸水平的改变可以使一系列细胞因子和酶表达水平改变,但是软骨细胞中骨桥蛋白高表达是否与透明质酸高表达相关目前尚不清楚。目的:通过调节体外培养人膝骨关节炎软骨细胞的骨桥蛋白水平,观察其对软骨细胞透明质酸表达水平的影响。方法:从人膝骨关节炎软骨组织标本中获取并体外培养软骨细胞,实验分为3组:空白对照组不进行任何干预,骨桥蛋白干预组以1 mg/L重组人骨桥蛋白干预,骨桥蛋白siR NA组以骨桥蛋白siR NA进行干预。采用real time PCR法检测各组骨桥蛋白,透明质酸合成酶1,透明质酸合成酶2和透明质酸合成酶3 mR NA表达,ELISA法检测透明质酸水平。结果与结论:(1)与空白对照组相比,重组人骨桥蛋白干预组透明质酸合成酶1,2,3表达升高,而骨桥蛋白siR NA干预组透明质酸合成酶1,2,3表达下降,3组间透明质酸合成酶1,2,3表达比较差异均有显著性意义(P<0.05);(2)重组人骨桥蛋白干预组软骨细胞透明质酸表达显著高于空白对照组和骨桥蛋白siR NA干预组(P<0.05)。(3)结果说明,提示骨桥蛋白可以通过刺激透明质酸合成酶分泌进而诱导骨关节炎软骨细胞透明质酸表达。

BACKGROUND：Both osteopontin and hyaluronic acid involve in the pathological process of osteoarthritis, resulting in the abnormal expression levels of various cytokines and enzymes. However, the relationship between the high expression of osteopontin and hyaluronic acid in chondrocytes remains unclear. OBJECTIVE：To investigate the effect of osteopontin on the expression of hyaluronic acid in human knee osteoarthritic chondrocytes in vitro by modulating the level of osteopontin. METHODS：Chondrocytes from human knee osteoarthritic cartilage were cultured in vitro, and were then divided into three groups：blank control group without any treatment;osteopontin group and and pontin siRNA group were treated with 1 mg/L recombinant human osteopontin and osteopontin siRNA, respectively. Expression levels of osteopontin, hyaluronic acid synthase 1, 2 and 3 mRNA were detected by real-time PCR, and the levels of hyaluronic acid were measured using ELISA. RESULTS AND CONCLUSION：Compared with the blank control group, the mRNA expressions of hyaluronic acid synthase 1, 2 and 3 were remarkably increased in the osteopontin group, while siRNA made the significantly inhibitory effects on the hyaluronic acid synthase 1, 2 and 3 mRNA expressions （P〈0.05）. The level of hyaluronic acid in chondrocytes in the osteopontin group was significantly higher than that in the other two groups （P〈0.05）. Our results suggest that osteopontin induces the synthesis of hyaluronic acid in osteoarthritic chondrocytes through upregulating the hyaluronic acid synthases expression levels.