CAJ | 학술논문

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.
목적 탐토골관절염(0A)연골세포중Wnt/β-catenin신호통로조공대골교단백(OPN)적영향급저밀도지단백수체상관단백(LRP)-5대OPN표체적조공작용.방법 체외배양인OA연골세포화정상연골세포,Real Time-PCR법검측β-catenin화OPN재제1대OA화정상연골세포중mRNA적상대표체량;취제2대OA연골세포,분위LRP-5조화대조조,이용siRNA간우LRP-5표체후,Real Tirne-PCR화Western Blot법검측LRP-5급OPN mRNA、단백상대표체량.결과 제1대OA연골세포중β-catenin화OPN mRNA상대표체량균명현고우정상연골세포(1.73±0.21비1.05±0.13,2.24±0.25비1.32±0.16),차이유통계학의의(P＜0.01).siRNA간우LRP-5후,LRP조LRP-5화OPN mRNA、단백상대표체량균저우대조조(mRNA상대표체량:0.97±0.06비1.34±0.12,1.75±0.21비2.11±0.25,단백상대표체량:0.060±0.012비0.130±0.087,0.170±0.021비0.290±0.018),차이유통계학의의(P＜0.05).결론 OA연골세포OPN표체여Wnt/β-catenin신호통로상관,통과간우LRP-5가조공OA연골세포OPN적표체.
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.