骨关节炎关节软骨自然退变过程中Dvl2及β-catenin表达变化  

Expression of Dvl2 and β-Catenin in Natural Degeneration Process of Osteoarthritis Articular Cartilage

作　　者：刘兴振[1] 范洁[2] 李甲[3] 綦柯[3] 李大河[3] 徐卫东[3] 赵东宝[1] LIU Xingzhen FAN Jie LI Jia QI Ke LI Dahe XU Weidong ZHAO Xa Dongbao(Changhai Hospital, The Second Military Medical University Department of Rheumatology, ＆Department of Joint Surgery, Shanghai 200433, China Department of Psychology, Hangzhou Sanatorium of PLA, Hangzhou 310007, China)

机构地区：[1]第二军医大学附属长海医院风湿免疫科,上海200433 [2]解放军杭州疗养院,杭州310007 [3]第二军医大学附属长海医院关节外科,上海200433

出　　处：《中国现代应用药学》2016年第11期1355-1358,共4页Chinese Journal of Modern Applied Pharmacy

基　　金：国家自然科学基金(81273284;81471607;81671595)

摘　　要：目的探讨Dishevelled(Dvl)和β-catenin在骨关节炎(osteoarthritis,OA)关节软骨细胞自然退变中的表达情况。方法将膝关节置换的OA患者关节软骨细胞进行体外分离培养,采用自然传代的方法建立关节软骨退变模型。倒置显微镜观察P2、P6代关节软骨细胞的形态,流式细胞仪检测软骨细胞的凋亡情况。利用Q-PCR及Western blot法检测Dvl2和β-catenin基因和蛋白的表达情况。结果成功分离并进行体外培养OA患者关节软骨细胞。P2代关节软骨细胞呈三角形或多角形;P6代时形态则变为梭形,并出现明显的纤维化。P6代时凋亡率(16.34%)显著高于P2代的凋亡率(8.63%)。P2代Dvl2的m RNA相对表达量为P6代的3.629倍(P=0.004),蛋白表达量为P6代的2.548倍(P=0.03),β-catenin的m RNA相对表达量为P6代的5.087倍(P=0.001),蛋白表达量为P6代的4.4倍(P=0.003)。结论 OA关节软骨细胞在体外自然退变过程中,凋亡增多,Dvl2及β-catenin表达则呈下降趋势。Dvl及Wnt/β-catenin在软骨细胞中的功能有待进一步研究。OBJECTIVE To investigate the expression changes of dishevelled（Dvl） and β-catenin in natural degeneration process of osteoarthritis（OA） articular cartilage. METHODS The articular chondrocytes from the patients with OA who underwent knee replacement were isolated and cultured in vitro. Using natural passage way to create a model of articular cartilage degeneration. The morphological of articular cartilage cells were observed under inverted microscope and the apoptosis of chondrocytes were detected by flow cytometry. Use Q-PCR and Western blot to test the genes and proteins expression of Dvl2 and β-catenin. RESULTS Successfully isolated and cultured OA articular chondrocytes in vitro. The articular chondrocytes of P2 generation was triangular or polygonal; morphological was fusiform of P6 generation, and appeared significant fibrosis. Apoptosis rate of P6 generation was significantly higher than P2 generation（16.34% vs 8.63%）. P2 generation＇s Dvl2 m RNA relative expression levels was P6 generation＇s 3.629-fold（P=0.004）, and protein expression levels was 2.548-fold（P=0.03）; β-catenin m RNA relative expression levels was P6 generation＇s 5.087-fold（P=0.001）, and protein expression levels was 4.4-fold（P=0.003）. CONCLUSION In the process of OA articular chondrocytes natural degeneration in vitro, apoptosis is increased and Dvl2 and β-catenin expression are declining. The function of Dvl and Wnt/β-catenin in chondrocytes requires further study.

关 键 词：WNT通路 骨性关节炎 软骨细胞 Β-CATENIN Dishevelled2 

分 类 号：R965.1[医药卫生—药理学]