摘要
目的观察人膝关节不同退变阶段骨关节炎(OA)软骨细胞中骨形态发生蛋白(BMP)信号通路相关分子的表达变化情况。方法根据Kellgren-Lawrence(K-L)X线分级膝关节软骨分为正常组(O级)、中期OA组(Ⅲ级)和晚期OA组(Ⅳ级)。各组收集软骨标本6~8例,体外分离与培养软骨细胞,采用第2代软骨细胞进行检测。运用实时定量PCR技术及Western-Blot技术检测BMP信号通路相关分子及软骨细胞表型标记物的表达情况。运用SPSS 22.0(IBM,USA)软件进行统计分析,计量资料采用方差分析,组间比较采用SNK法。结果随着OA退变程度加重,BMP信号通路中激活素受体样激酶1(ALK 1)和TGF-β家族信号通路的下游信号分子Smad 1表达升高(F=33.6,P<0.05;F=21.3,P<0.05),人性别决定区Y框蛋白9(Sox 9)转录因子表达下降(F=21.0,P<0.05),与正常软骨组比较均有统计学差异。而中期OA组与晚期OA组无统计差异(P>0.05);Smad 7表达在各组间差异无统计学意义(P>0.05)。随着OA退变程度加重,软骨细胞正常表型标记物Ⅱ型胶原表达降低(F=234.6,P<0.05),而肥大分化表型标记物X型胶原及金属蛋白酶3表达则升高,各组间差异均有统计学意义(F=81.0,P<0.05;F=29.8,P<0.05)。结论在OA软骨细胞中,BMP信号通路相关分子高表达是软骨细胞退变及OA的进展的重要因素。
Objective To investigate the expression of molecules of bone morphogenetic protein（ BMP） signaling pathway in different stage of osteoarthritis（ OA）. Methods Cartilage was classified into three groups,normal group,mid-staged group and advanced-staged group,according to the Kellgren and Lawrence criterion. Six to eight cases of normal or OA cartilage were collected and chondrocytes were isolated. The second generation of chondrocytes were used to examination. The levels of molecules of BMP signaling pathway mRNA and protein expressions were measured with real-time fluorescent quantitative PCR and Western-Blot. The statistical analysis was performed with SPSS 22. 0 software,the numerical variables were compared using the one-way analysis of variance among the groups and SNK method between the two groups. Results The expression of activin receptor-like kinase 1（ ALK1） and Smad1 increased significantly（ F = 33. 6,P〈0. 05; F = 21. 3,P〈0. 05） with degradation of chondrocytes,while the expression of SRY-related high mobility group-box gene（ Sox） 9 decreased significantly（ F = 21. 0,P〈0. 05）. The expression of Smad7 showed a decreased trend. The expression of type Ⅱ collagen（ COL2 A1）decreased significantly（ F = 234. 6,P〈0. 05） with degradation of chondrocytes,while the expression of type X collagen（ COL10 A1） and matrix metallo proteinase-3（ MMP-3） increased significantly（ F = 81. 0,P〈0. 05; F = 29. 8,P〈0. 05）. Conclusion Over expression of related molecules of BMP signaling pathway in articular chondrocytes leads to the articular cartilage degeneration and the development of OA.
出处
《中华关节外科杂志（电子版）》
CAS
2017年第5期50-54,共5页
Chinese Journal of Joint Surgery(Electronic Edition)
基金
国家自然科学基金项目(81672219
81601936)
四川省科技厅科技支撑计划(2014SZ0023-2)
