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| miR-92a-3p靶向EZH2对IL-1β诱导软骨细胞合成基质降解酶的影响 |
| Effect of miR-92a-3p targeting EZH2 on the synthesis of matrix degrading enzymes in chondrocytes induced by IL-1β |
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| DOI：10.3969/j.issn.1006-7108.2020.09.011 |
| 中文关键词: 软骨细胞 EZH2 miR-92a-3p 基质降解酶 |
| 英文关键词:chondrocyte EZH2 miR-92a-3p matrix degrading enzyme |
| 基金项目:武汉市卫生健康委员会资助项目（WX19C18） |
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| 中文摘要: |
| 目的 研究miR-92a-3p靶向zeste基因增强子同源物2（EZH2）对白细胞介素-1β（IL-1β）诱导的软骨细胞合成基质降解酶的影响。方法 分离培养人软骨细胞，分成Normal、IL-1β（IL-1β处理）、miR-NC+IL-1β（mimics control转染后IL-1β处理）、miR-92a-3p+IL-1β（miR-92a-3p mimics转染后IL-1β处理）组，RT-PCR方法测定miR-92a-3p表达变化，Western blot检测MMP-13、MMP-1、COLⅡ蛋白表达。靶基因预测软件发现EZH2可能为miR-92a-3p的靶基因，荧光素酶报告系统鉴定靶向关系。将pCDNA3.1-EZH2和miR-92a-3p mimics共同转染到软骨细胞中，检测MMP-13、MMP-1、COLⅡ蛋白表达情况。结果 与Normal比较，IL-1β组细胞中miR-92a-3p表达减少，MMP-13、MMP-1表达增多，COLⅡ表达减少（P<0.05）。与miR-NC+IL-1β比较，miR-92a-3p+IL-1β组细胞中miR-92a-3p表达增多，MMP-13、MMP-1表达减少，COLⅡ表达增多（P<0.05）。miR-92a-3p靶向负调控EZH2表达。pCDNA3.1-EZH2可以逆转miR-92a-3p mimics对IL-1β条件下软骨细胞中MMP-13、MMP-1、COLⅡ蛋白表达影响。结论 miR-92a-3p靶向下调EZH2抑制IL-1β诱导的软骨细胞合成基质降解酶。 |
| 英文摘要: |
| Objective To study the effect of miR-92a-3p targeting EZH2 on the synthesis of matrix degrading enzymes in chondrocytes induced by IL-1β. Methods Human chondrocytes were divided into normal, IL-1β (treatment of IL-1β), miR-NC+IL-1β (treatment of IL-1β after mimics control transfection), and miR-92a-3p+IL-1β (treatment of IL-1βafter miR-92a-3p mimics transfection) group. The expression of miR-92a-3p was determined using RT-PCR. Western blotting was used to detect the expression of MMP-13, MMP-1, and col Ⅱ. It was predicted with a software that EZH2 might be the target gene of miR-92a-3p. The target relationship was identified by luciferase report system. pCDNA3.1-EZH2 and miR-92a-3p mimics were co-transfected into chondrocytes. The protein expressions of MMP-13, MMP-1, and col Ⅱ were detected. Results Compared with those in normal group, the expression of miR-92a-3p decreased, the expressions of MMP-13 and MMP-1 increased, and the expression of col Ⅱ decreased in IL- β group (P< 0.05). Compared with those in miR-NC+ IL-1β group, the expression of miR-92a-3p increased, the expressions of MMP-13 and MMP-1 reduced, and the expression of col Ⅱ increased in miR-92a-3p + IL-1β group (P<0.05). miR-92a-3p negatively target-regulated EZH2 expression. pCDNA3.1-EZH2 reversed the effect of miR-92a-3p mimics on the expressions of MMP-13, MMP-1, and col Ⅱ in chondrocytes with the condition of IL-1β. Conclusion miR-92a-3p targeted down-regulation of EZH2 inhibits IL-1β- induced matrix degrading enzyme synthesis in chondrocytes. |
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