| 摘要: |
| 目的 观察早期骨关节炎（osteoarthritis，OA）软骨细胞中miRNA-140的表达规律以及转染ds-miRNA-140对软骨细胞功能的影响。 方法 建立兔OA模型，获得正常软骨细胞（A组）和4周（B组）、8周（C组）OA软骨细胞，应用实时定量PCR和蛋白质印迹法检测各组miRNA-140相对表达量以及Ⅱ型胶原蛋白（Col2a1）和金属蛋白酶13（MMP-13）的表达情况。各组软骨细胞转染ds-miRNA-140后，观察细胞Col2a1、MMP-13的表达情况。 结果 B组、C组软骨细胞miRNA-140的表达和A组相比分别下降到63%和57%（P<0.01），而B组和C组软骨细胞miRNA-140的表达差异无统计学意义（P>0.05）；B、C两组细胞Col2a1 mRNA的表达和A组相比分别减少了52%和63%（P<0.01），B、C两组软骨细胞MMP-13 mRNA的表达和A组相比分别升高了3.01倍和4.15倍（P<0.01）。转染ds-miRNA-140后，B、C两组细胞Col2a1 mRNA的表达分别增加了60%和127%（P<0.01），B、C两组MMP-13 mRNA的表达在转染后是转染前的54.53%和42.61%（P<0.01）。Col2a1和MMP-13蛋白表达水平变化趋势与mRNA一致。 结论 OA早期关节软骨miRNA-140的表达显著减少；转染ds-miRNA-140可增加软骨细胞Col2a1表达，同时降低MMP-13的表达。 |
| 关键词: 骨关节炎 miRNA-140 软骨细胞 胶原Ⅱ型 基质金属蛋白酶13 |
| DOI：10.3724/SP.J.1008.2014.00610 |
| 投稿时间：2013-11-12修订日期：2014-05-19 |
| 基金项目:国家重点基础研究规划("973"计划) 2012CB619105 |
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| Expression of miRNA-140 in chondrocytes of patients with early osteoarthritis and its function |
| ZHOU Xin-hua1,WANG Min2*,JI Yan-hui2,JIANG Yi1,LIU Qing1 |
(1. Department of Orthopaedics, Beijing Jishuitan Hospital, The Fourth Clinical College of Peking University, Beijing 100035, China;
2. Department of Orthopaedics, Xinqiao Hospital, Third Military Medical University, Chongqing 400037, China
*Corresponding authors.) |
| Abstract: |
| Objective To observe miRNA-140 expression in chondrocytes of patients with early osteoarthritis (OA)and the effect of transfecting double-stranded miR-140 (ds-miRNA-140) on chondrocyte function. Methods Normal and OA chondrocytes (4 weeks and 8 weeks) were collected from rabbit model of early OA (A, B and C group, respectively). Quantitative polymerase chain reaction was used to examine miRNA-140 expression and Western blotting analysis was used to examine Col2a1 and MMP-13 protein expression in each group. Col2a1 and MMP-13 protein expression was also observed in chondrocytes transfected with ds-miR-140. Results Compared with group A, miR-140 expressions in group B and C were reduced to 63% and 57%, respectively (P<0.01); the expression was not significantly different between the group B and C(P>0.05). Compared with group A, Col2a1 mRNA expressions in group B and C were reduced by 52% and 63%, respectively (P<0.01); while MMP-13 mRNA expressions were up-regulated by 3.01 and 4.15 folds, respectively (P<0.01). Transfection with ds-miR-140 increased Col2a1 mRNA by 60% and 127% in group B and C, respectively(P<0.01), and the expressions of MMP-13 mRNA in group B and C were reduced to 54.53% and 42.61%, respectively(P<0.01). The changes of Col2a1 and MMP-13 protein expression were the same as that of mPNA. Conclusion miR-140 expression is reduced in early OA cartilage, and transfection with ds-miR-140 may increase Col2a1 expression and reduce MMP-13 expression. |
| Key words: osteoarthritis miRNA-140 chondrocytes collagen tyep Ⅱ matrix metalloproteinase 13 |
