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Col2a1mRNA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-13mRNA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Col2a1mRNA的表达分别增加了60%和127%(P<0.01),B、C两组MMP-13mRNA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。
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 Col2al and MMP-13 protein expression in each group. Co12al 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, Col2al 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 Col2al 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 Col2al 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 Col2al expression and reduce MMP-13 expression.
出处
《第二军医大学学报》
CAS
CSCD
北大核心
2014年第6期610-615,共6页
Academic Journal of Second Military Medical University
