Si Haibo, Zeng Yi, Zhou Zongke, Pei Fuxing, Lu Yanrong, Cheng Jingqiu, Shen Bin
Department of Orthopedics Regenerative Medicine Research Centre, West China Hospital, Sichuan University, Chengdu 610041, China; Key Laboratory of Transplant Engineering and Immunology, Regenerative Medicine Research Centre, West China Hospital, Sichuan University, Chengdu 610041, China.
Department of Orthopedics, Regenerative Medicine Research Centre, West China Hospital, Sichuan University, Chengdu 610041, China.
Chin Med Sci J. 2016 Nov 20;31(4):207-212. doi: 10.1016/s1001-9294(17)30002-0.
Objective To investigate the expression of miRNA-140 in chondrocytes and synovial fluid of osteoarthritis (OA) patients, and explore the relationship between the miRNA-140 expression and OA severity.Methods This study enrolled 30 OA patients who underwent total knee arthroplasty for chondrocytes sampling and 30 OA patients who underwent intra-articular injection for synovial fluid sampling. All OA patients were grouped into mild [Kellgren and Lawrence (KL) grade 1-2], moderate (KL grade 3) and severe (KL grade 4), with 10 in each subgroups for each sampling purposes. 7 non-OA patients and 10 patients with knee injury were collected for cartilage and synovial fluid sampling respectively as control groups. Chondrocytes were isolated from the cartilage tissue and cultured in vitro. Quantitative real time PCR for miRNA-140 in chondrocytes and synovial fluid were performed, and the U6 snRNA was used as internal control. The expression difference of miRNA-140 among groups and correlation between the expression and the KL grade of OA were analysed using one-way ANOVA and Spearman test respectively. Results The expression of miRNA-140 in chondrocytes of knees in OA patients was reduced than that in normal knees, and the between-group difference was statistically significant (F=305.464, P<0.001). miRNA-140 could be detected in synovial fluid of both normal knees and OA knees, its relative expression level was reduced in synovial fluid of OA group compared with normal group, and the between-group difference was statistically significant as well (F=314.245, P<0.001). The relative expression level of miRNA-140 in both chondrocytes and synovial fluid were negatively correlated with the KL grade of OA(r=-0.969, P<0.001; r=-0.970, P<0.001). Conclusion miRNA-140 could be detected in chondrocytes and synovial fluid of OA patients, and its expression was negatively correlated with the severity of OA.
目的 探讨微小RNA-140（miRNA-140）在骨关节炎（OA）患者软骨细胞及滑液中的表达情况，并探究miRNA-140表达与OA严重程度之间的关系。方法 本研究纳入30例行全膝关节置换术以获取软骨细胞样本的OA患者和30例行关节腔内注射以获取滑液样本的OA患者。所有OA患者分为轻度[凯尔格伦和劳伦斯（KL）分级1-2级]、中度（KL分级3级）和重度（KL分级4级），每种采样目的各亚组均为10例。分别收集7例非OA患者和10例膝关节损伤患者的软骨和滑液样本作为对照组。从软骨组织中分离软骨细胞并进行体外培养。对软骨细胞和滑液中的miRNA-140进行实时定量聚合酶链反应（PCR），以U6小核RNA（snRNA）作为内参。分别采用单因素方差分析和Spearman检验分析各组间miRNA-140的表达差异以及其表达与OA的KL分级之间的相关性。结果 OA患者膝关节软骨细胞中miRNA-140的表达低于正常膝关节，组间差异具有统计学意义（F=305.464，P<0.001）。正常膝关节和OA膝关节的滑液中均可检测到miRNA-140，OA组滑液中其相对表达水平低于正常组，组间差异也具有统计学意义（F=314.245，P<0.001）。软骨细胞和滑液中miRNA-140的相对表达水平均与OA的KL分级呈负相关（r=-0.969，P<0.001；r=-0.970，P<0.001）。结论 OA患者的软骨细胞和滑液中均可检测到miRNA-140，其表达与OA严重程度呈负相关。
