摘要
目的 探讨骨关节炎（0A）软骨细胞中Wnt/β-catenin信号通路调控对骨桥蛋白（OPN）的影响及低密度脂蛋白受体相关蛋白（LRP）-5对OPN表达的调控作用.方法 体外培养人OA软骨细胞和正常软骨细胞,Real Time-PCR法检测β-catenin和OPN在第1代OA和正常软骨细胞中mRNA的相对表达量;取第2代OA软骨细胞,分为LRP-5组和对照组,利用siRNA干扰LRP-5表达后,Real Tirne-PCR和Western Blot法检测LRP-5及OPN mRNA、蛋白相对表达量.结果 第1代OA软骨细胞中β-catenin和OPN mRNA相对表达量均明显高于正常软骨细胞（1.73±0.21比1.05±0.13,2.24±0.25比1.32±0.16）,差异有统计学意义（P＜0.01）.siRNA干扰LRP-5后,LRP组LRP-5和OPN mRNA、蛋白相对表达量均低于对照组（mRNA相对表达量：0.97±0.06比1.34±0.12,1.75±0.21比2.11±0.25,蛋白相对表达量：0.060±0.012比0.130±0.087,0.170±0.021比0.290±0.018）,差异有统计学意义（P＜0.05）.结论 OA软骨细胞OPN表达与Wnt/β-catenin信号通路相关,通过干扰LRP-5可调控OA软骨细胞OPN的表达.
Objective To investigate the effect of Wnt/ β-catenin signaling pathway on osteopontin （OPN） in osteoarthritis （OA） chondrocytes and low-density lipoprotein receptor-related protein （LRP）-5 on the regulation of expression of OPN.Methods Primary cultured human OA and normal chondrocytes,the expression of β-catenin and OPN mRNA were investigated by Real Time-PCR in the first generation of OA and normal chondrocytes.Taken the second generation of OA chondrocytes,blocking LRP-5 expression was performed using small interfering RNA（siRNA） in OA chondrocytes.Then the mRNA and protein expression levels of LPR-5 and OPN were measured by the Real Time-PCR and Western Blot.Results The expression of β-catenin and OPN mRNA in OA chondrocytes were significantly higher than those in normal chondrocytes （1.73 ± 0.21 vs.1.05 ± 0.13,2.24 ± 0.25 vs.1.32 ± 0.16,P ＜ 0.01）.After siRNA interfering with LRP-5,the expression of LRP-5 and OPN mRNA and protein in OA chondrocytes were significantly lower than those in normal chondrocytes （the expression of mRNA：0.97 ± 0.06 vs.1.34 ± 0.12,1.75 ± 0.21 vs.2.11 ±0.25; the expression of protein：0.060 ±0.012 vs.0.130 ±0.087,0.170 ±0.021 vs.0.290 ± 0.018,P ＜ 0.01）.Conclusions OPN expression is related to Wnt/3-catenin signaling pathway in OA chondrocytes,by interfering with LRP-5 that can regulate expression of OPN in OA chondrocytes.
出处
《中国医师进修杂志》
2014年第5期1-4,共4页
Chinese Journal of Postgraduates of Medicine
基金
温州市科技局资助项目（Y20110116）
