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。
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-1β after 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-1βgroup(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.
作者
张明焕
毛文
刘雷
祁福
李北
郑一舟
ZHANG Minghuan;MAO Wen;LIU Lei;QI Fu;LI Bei;ZHENG Yizhou(Department of Orthopedics,Wuhan No.3 Hospital,Wuhan 430071,China)
出处
《中国骨质疏松杂志》
CAS
CSCD
北大核心
2020年第9期1301-1305,1363,共6页
Chinese Journal of Osteoporosis
基金
武汉市卫生健康委员会资助项目(WX19C18)。
