摘要
目的检测骨关节炎(OA)患者骨桥蛋白(OPN)的表达,探讨Wnt/β-catenin信号通路对OPN的表达调控作用。方法分离OA患者(骨关节炎组12例)的软骨细胞,并取非OA患者软骨细胞作为正常对照(8例),荧光定量PCR检测OPN基因的表达;基因转染双链siRNA干扰β-catenin基因的表达;Western blot实验检测β-catenin和OPN蛋白的表达改变。结果骨关节炎组和正常对照组软骨细胞OPN的mRNA表达量分别为(0.642±0.155)和(0.226±0.077),两组比较差异有统计学意义(t=6.74,P<0.01)。特异性靶向双链siRNA可以下调β-catenin的表达,Western blot检测显示下调β-catenin蛋白的表达后软骨细胞OPN蛋白的表达也出现了相应的下调。结论骨关节炎患者中存在骨桥蛋白的表达上调,骨桥蛋白的上调受到Wnt/β-catenin通路的调控。
Objective To detect the expression of osteopontin （OPN） in osteoarthritis （OA） patients, and explore the regulation mechanism of OPN by Wnt/β-catenin signaling pathway. Methods The chondrocytes were separated from OA patients （12 cases as OA group） and non-OA patients （8 cases as control group）. The mRNA expression of OPN was determined by real-time PCR analysis. The small interfering RNA was used to specifically knockdown β-catenin in chondrocytes. And then the expression of β-catenin and OPN protein were detected by Western blot. Results The 2-△Ct values ofOPN mRNA were （0.226±0.077） and （0.642±0.155） in control and OA group, respectively. There was significant difference between the two groups （t=6.74,P〈0.01）. The expression of β-catenin was significantly and specifically down-regulated by siRNA duplexes and a corresponding down-regulation was also detected in OPN protein expression by Western blot. Conclusion There is an up-regulated expression of OPN in osteoarthritis patients, which is mediated by Wnt/β-catenin signaling pathway.
出处
《海南医学》
CAS
2014年第13期1877-1879,共3页
Hainan Medical Journal
