DDR2调控LncRNA UCA1/miR-590-3p轴对骨关节炎软骨细胞炎症因子释放和MMPs表达的影响  

Effects of DDR2 regulation of LncRNA UCA1/miR-590-3p axis on the release of inflammatory factors and the expression of MMPs in osteoarthritis chondrocytes

作　　者：孙强 张大伟 陈永锋 王华溢 王远瑞 王鹏 宋和强 贾鸿 SUN Qiang;ZHANG Da-wei;CHEN Yong-feng;WANG Hua-yi;WANG Yuan-rui;WANG Peng;SONG He-qiang;JIA Hong(Department of Orthopedic Surgery,Xijing Hospital,Air Force Medical University,Xi’an,Shaanxi,710032,China)

机构地区：[1]中国人民解放军空军军医大学第一附属医院骨科,陕西710032

出　　处：《中国骨与关节杂志》2023年第4期278-284,共7页Chinese Journal of Bone and Joint

基　　金：陕西省重点研发计划(2022SF-229)。

摘　　要：目的探讨盘状结构域受体(discoidin domain receptor,DDR2)调控长链非编码RNA-尿路上皮癌相关分子1(long noncoding RNA-urothelial carcinoma antigen 1,LncRNA UCA1)/miR-590-3p轴对骨关节炎(osteoarthritis,OA)软骨细胞炎症因子释放和基质金属蛋白酶(matrix metalloproteinases,MMPs)表达的影响及机制。方法采用RT-q PCR检测DDR2、LncRNA UCA1和miR-590-3p在OA软骨细胞和正常软骨细胞中的表达水平;在OA软骨细胞中分别转染sh-UCA1、miR-590-3p mimics、LV-UCA1和LV-UCA1+miR-590-3p mimics后实时荧光定量聚合酶链反应(real-time quantitative polymerase chain reaction,RT-qPCR)检测白介素-6(interleukin-6,IL-6)和肿瘤坏死因子-α(tumor necrosis factorα,TNF-α)的表达水平,RT-qPCR和蛋白质印迹(western blot,WB)检测MMP-3和MMP-13的表达水平。双荧光素酶基因报告检测LncRNA UCA1和miR-590-3p的结合情况;RNA免疫共沉淀(RNA immunoprecipitation,RIP)检测LncRNA UCA1与miR-590-3p和Argonaut-2蛋白(Ago-2)的结合情况。结果在OA患者软骨细胞中DDR2和LncRNA UCA1的表达水平明显高于正常软骨细胞,miR-590-3p的表达水平明显低于正常软骨细胞。DDR2能够促进LncRNA UCA1的表达和抑制miR-590-3p的表达。双荧光素酶基因报告和RIP结果显示LncRNA UCA1能够通过ceRNA机制靶向抑制miR-590-3p的表达。低表达LncRNA UCA1和过表达miR-590-3p能够显著抑制IL-6、TNF-α、MMP-3和MMP-13的表达水平。过表达LncRNA UCA1能够显著促进IL-6、TNF-α、MMP-3和MMP-13的表达水平,同时过表达UCA1和miR-590-3p能够反转单独过表达LncRNA UCA1对OA软骨细胞炎症因子释放和MMPs表达的影响。结论DDR2能够上调LncRNA UCA1,LncRNA UCA1通过ceRNA机制靶向抑制miR-590-3p,进而促进OA软骨细胞炎症因子释放和MMPs的表达。Objective To investigate the effect and mechanism of DDR2-regulated LncRNA UCA1/miR-590-3p axis on the release of inflammatory factors and the expression of matrix metalloproteinases(MMPs)in chondrocytes of osteoarthritis(OA).Methods The expression levels of DDR2,LncRNA UCA1 and miR-590-3p in OA chondrocytes and normal chondrocytes were detected by RT-qPCR.After transfection of sh-UCA1,miR-590-3p mimics,LV-UCA1 and LV-UCA1+miR-590-3p mimics in OA chondrocytes,the expression levels of IL-6 and TNF-αwere detected by RT-qPCR,and the expression levels of MMP-3 and MMP-13 were detected by RT-qPCR and WB.The binding of LncRNA UCA1 and miR-590-3p was detected by dual luciferase gene reports.RIP was used to detect the binding of LncRNA UCA1 and miR-59.Results The expression levels of DDR2 and LncRNA UCA1 in chondrocytes of patients with OA were significantly higher than those in normal chondrocytes,and the expression level of miR-590-3p was significantly lower than that in normal chondrocytes.DDR2 could promote the expression of LncRNA UCA1 and inhibit the expression of miR-590-3p.The dual-luciferase gene reporter and RIP results showed that the LncRNA UCA1 could target the expression of miR-590-3p through a ceRNA mechanism.Low expression of LncRNA UCA1 and overexpression of miR-590-3p could significantly inhibit the expression levels of IL-6,TNF-α,MMP-3 and MMP-13.Overexpression of LncRNA UCA1 can significantly promote the expression levels of IL-6,TNF-α,MMP-3 and MMP-13,while overexpression of UCA1 and miR-590-3p could reverse the effect of overexpression of LncRNA UCA1 alone on inflammatory factors release and the expression of MMPs in OA chondrocytes.Conclusions DDR2 can up-regulate LncRNA UCA1,which targets miR-590-3p through a ceRNA mechanism,thereby promoting the release of inflammatory factors and the expression of MMPs in OA chondrocytes.

关 键 词：骨关节炎 盘状结构域受体2 RNA 长链非编码 基质金属蛋白酶 炎症因子 

分 类 号：R684.3[医药卫生—骨科学]