miR-140-5p通过NF-κB通路调控人骨关节炎软骨细胞自噬的研究  

miRNA-140-5p regulates chondrocyte autophagy in human osteoarthritis through the NF-κB pathway

作　　者：刘志刚 李琦军[2] 王飞[3] 翟羽佳 LIU Zhigang;LI Qijun;WANG Fei;ZHAI Yujia(The People’s Hospital of Zhengding County, Zhengding 050800,Hebei, China;The Third Hospital of Shijiazhuang, Shijiazhuang 050000, Hebei, China;The Third Hospital of Hebei Medical University, Shijiazhuang 050000, Hebei, China)

机构地区：[1]河北省正定县人民医院,河北正定050800 [2]河北省石家庄市第三医院,河北石家庄050000 [3]河北医科大学附属第三医院,河北石家庄050000

出　　处：《现代中西医结合杂志》2020年第19期2089-2093,共5页Modern Journal of Integrated Traditional Chinese and Western Medicine

摘　　要：目的探讨miR-140-5p在正常和骨关节炎软骨细胞中的表达情况及其在骨关节炎发病机制中的作用。方法①通过定量实时聚合酶链反应(RT-qPCR)检测骨关节炎软骨细胞和正常软骨细胞中miR-140-5p和基质金属蛋白酶13(MMP-13)表达水平。②将软骨细胞分为3组,对照组加入白细胞介素-1β(IL-1β)培养,miR-140-5p mimic组同时转染miR-140-5p mimic,miR-140-5p inhibitor组同时转染miR-140-5p inhibitor。采用MTT法检测各组软骨细胞增殖情况,采用RT-qPCR和Western blot法检测各组MMP-13、Beclin-1、ATG7和NF-κB信号通路中NF-κBp65表达情况。结果骨关节炎软骨细胞中miR-140-5p相对表达水平较正常软骨细胞中显著降低(P<0.05),MMP-13相对表达水平较正常软骨细胞中显著增高(P<0.05)。与对照组比较,miR-140-5p mimic组软骨细胞增殖明显,miR-140-5p inhibitor组软骨细胞增殖明显受到抑制。与对照组比较,miR-140-5p mimic组软骨细胞中MMP-13、NF-κBp65mRNA和蛋白表达水平均明显降低(P均<0.05),而miR-140-5p inhibitor组均明显升高(P均<0.05);miR-140-5p mimic组软骨细胞中Beclin-1和ATG7mRNA和蛋白表达水平均明显升高(P均<0.05),而miR-140-5p inhibitor组均明显降低(P均<0.05)。结论miR-140-5p能够通过抑制NF-κB信号通路促进骨关节炎软骨细胞的增殖和自噬,延缓骨关节炎的进展,为探索靶向miR-140-5p治疗骨关节炎提供了一定参考。Objective It is to explore the expression of miRNA-140-5p in normal and osteoarthritis chondrocytes and its role in the pathogenesis of osteoarthritis.Methods①The expression levels of miR-140-5p and matrix metalloproteinase 13(MMP-13)in osteoarthritis chondrocytes and normal chondrocytes were detected by quantitative real-time polymerase chain reaction(RT-qPCR).②The chondrocytes were divided into 3 groups,the control group was added with interleukin-1β(IL-1β)for culture,miR-140-5p mimic group was simultaneously transfected with miR-140-5p mimic,miR-140-5p inhibitor group was simultaneously transfected with miR-140-5p inhibitor.MTT method was used to detect the proliferation of chondrocytes in each group,and RT-qPCR and Western blot were used to detect the expression of MMP-13,Beclin-1,ATG7and NF-κBp65 in NF-κB signaling pathways in each group.Results The relative expression level of miR-140-5p in osteoarthritis chondrocytes was significantly lower than that in normal chondrocytes(P<0.05),and the relative expression level of MMP-13 was significantly higher than that in normal chondrocytes(P<0.05).Compared with the control group,the proliferation of chondrocytes in the miR-140-5p mimic group was significant,and the proliferation of the chondrocytes in the miR-140-5p inhibitor group was significantly inhibited.Compared with the control group,the expression levels of MMP-13,NF-κBp65 mRNA and protein in chondrocytes of miR-140-5p mimic group were significantly reduced(P<0.05),while the expression in miR-140-5p inhibitor group were significantly increased(P<0.05);the expression levels of Beclin-1 and ATG7mRNA and protein chondrocytes in miR-140-5p mimic group were significantly increased(P<0.05),while the expression in miR-140-5p inhibitor group were significantly reduced(P<0.05).Conclusion MiR-140-5p can promote the proliferation and autophagy of osteoarthritis chondrocytes by inhibiting the NF-κB signaling pathway,thus to delay the progress of osteoarthritis.The result provides a reference for exploring the

关 键 词：骨关节炎 自噬 miR-140-5p NF-ΚB信号通路 

分 类 号：R-33[医药卫生]