lncRNA SNHG14靶向miR-181a-5p/PRDX3轴对IL-1β诱导的软骨细胞损伤的影响

doi:10.3870/j.issn.1672-8009.2026.02.002

摘要/Abstract

摘要： 目的探讨长链非编码RNA小核仁RNA宿主基因14(long non-coding rna small nucleolar RNA host gene 14,lncRNA SNHG14)靶向微小RNA-181a-5p(microRNA-181a-5p,miR-181a-5p)/过氧化物还原酶3(peroxiredoxin 3,PRDX3)轴对白细胞介素(interleukin,IL)-1β诱导的软骨细胞损伤的影响。方法双荧光素酶实验检测lncRNA SNHG14、miR-181a-5p和PRDX3靶向关系。人软骨细胞C28/I2分为以下10组:NC组、IL-1β组、sh-NC组、sh-SNHG14组、sh-SNHG14+anti-NC组、sh-SNHG14+anti-miR-181a-5p组、miR-NC组、miR-181a-5p mimic组、miR-181a-5p mimic+pcDNA-NC组、miR-181a-5p mimic+pcDNA-PRDX3组。克隆形成实验、流式细胞术分别检测细胞增殖和凋亡情况;酶联吸附免疫法检测IL-6、肿瘤坏死因子α(tumor necrosis factor alpha,TNF-α)、单核细胞趋化蛋白1(monocyte chemoattractant protein 1,MCP-1)水平;Griess试剂法检测一氧化氮(nitric oxide,NO)水平;蛋白质印迹检测PRDX3、增殖细胞核抗原(proliferating cell nuclear antigen,PCNA)、细胞周期蛋白(Cyclin D1)、B细胞淋巴瘤/白血病-2(B-cell lymphoma 2,Bcl-2)、Bcl-2家族相关X蛋白(Bcl2-associated X protein,Bax)蛋白表达量。结果双荧光素酶实验结果显示miR-181a-5p与lncRNA SNHG14、PRDX3均存在靶向关系(P<0.05)。与NC组比较,IL-1β组细胞凋亡率,IL-6,TNF-α,MCP-1,NO水平,lncRNA SNHG14表达量,PRDX3,Bax蛋白表达量明显上升,miR-181a-5p表达量,集落形成数量,PCNA、Cyclin D1、Bcl-2蛋白表达水平明显下降(P<0.05);与sh-NC组或miR-NC组相比,sh-SNHG14组或miR-181a-5p mimic组细胞中miR-181a-5p表达量,集落形成数量,PCNA、Cyclin D1、Bcl-2蛋白表达水平明显升高,细胞凋亡率,IL-6、TNF-α、MCP-1、NO水平,lncRNA SNHG14表达量,PRDX3和Bax蛋白表达量明显降低(P<0.05);与sh-SNHG14+anti-NC组比较,sh-SNHG14+anti-miR-181a-5p组细胞中各项检测指标变化趋势与之相反(P<0.05);而miR-181a-5p mimic+pcDNA-PRDX3组细胞的各指标趋势均与miR-181a-5p mimic+pcDNA-NC组相反(P<0.05)。结论 lncRNA SNHG14通过靶向miR-181a-5p/PRDX3轴缓解IL-1β诱导的软骨细胞损伤。

关键词: 小核仁RNA宿主基因14, 微小RNA-181a-5p, 过氧化物还原酶3, 骨关节炎

Abstract: Objective To explore the effect of long non coding RNA small nucleolar RNA host gene 14(lncRNA SNHG14)on interleukin(IL)-1β-induced chondrocyte injury by targeting the microRNA-181a-5p(miR-181a-5p)/peroxiredoxin3 (PRDX3) axis.Methods The targeting relationships among lncRNA SNHG14,miR-181a-5p,and PRDX3 were detected using dual-luciferase reporter assay.Human chondrocytes C28/I2 were used to set up the following 10 groups:NC group,IL-1β group,sh-NC group,sh-SNHG14 group,sh-SNHG14+anti-NC group,sh-SNHG14+anti-miR-181a-5p group,miR-NC group,miR-181a-5p mimic group,miR-181a-5p mimic+pcDNA-NC group,miR-181a-5p mimic+pcDNA-PRDX3 group.Cell proliferation and apoptosis were assessed using colony formation assay and flow cytometry,respectively.The levels of IL-6,tumor necrosis factor-alpha(TNF-α),and monocyte chemoattractant protein 1(MCP-1)were measured by enzyme-linked immunosorbent assay.Nitric oxide(NO)level was detected using Griess reagent.Western blotting was performed to determine the expression levels of PRDX3,proliferating cell nuclear antigen(PCNA),Cyclin D1,B-cell lymphoma 2(Bcl-2),and Bcl-2-associated X protein(Bax).Results Dual luciferase assay indicated the targeting relationships between miR-181a-5p and both lncRNA SNHG14 and PRDX3(P<0.05).Compared with the NC group,the IL-1β group showed significantly increased apoptosis rate,IL-6,TNF-α,MCP-1,and NO levels,as well as elevated expression levels of lncRNA SNHG14,PRDX3,and Bax protein,and decreased miR-181a-5p expression,colony formation number,and protein levels of PCNA,Cyclin D1,and Bcl-2(P<0.05).Compared with the sh-NC group or miR-NC group,the sh-SNHG14 group or miR-181a-5p mimic group exhibited significantly higher miR-181a-5p expression level,colony formation number,and protein expression level of PCNA,Cyclin D1,and Bcl-2,along with notably reduced apoptosis rate,IL-6,TNF-α,MCP-1,and NO levels,as well as decreased expression levels of lncRNA SNHG14,PRDX3,and Bax protein(P<0.05).In contrast to the sh-SNHG14+anti-NC group,the sh-SNHG14+anti-miR-181a-5p group demonstrated opposite trends in all above detected indicators(P<0.05).Similarly,all trends of the above index in the miR-181a-5p mimic+pcDNA-PRDX3 group were reversed compared with those in the miR-181a-5p mimic+pcDNA-NC group(P<0.05).Conclusion LncRNA SNHG14 alleviates IL-1β-induced chondrocyte injury by targeting the miR-181a-5p/PRDX3 axis.

Key words: small nucleolar RNA host gene 14, microRNA-181a-5p, peroxiredoxin 3, osteoarthritis

中图分类号:

R349.7

宋凯林, 罗燕宁, 方红育. lncRNA SNHG14靶向miR-181a-5p/PRDX3轴对IL-1β诱导的软骨细胞损伤的影响[J]. 医学分子生物学杂志, 2026, 23(2): 116-126.

SONG Kailin, LUO Yanning, FANG Hongyu. Effect of LncRNA SNHG14 on IL-1β-Induced Chondrocyte Injury by Targeting miR-181a-5p/PRDX3 Axis[J]. Journal of Medical Molecular Biology, 2026, 23(2): 116-126.

参考文献

[1] MOTTA F,BARONE E,SICA A,et al.Inflammaging and osteoarthritis[J].Clin Rev Allergy Immunol,2023,64(2):222-238.
[2] TANG S A,ZHANG C,OO W M,et al.Osteoarthritis[J].Nat Rev Dis Primers,2025,11:10.
[3] WANG Y,FAN X,XING L,et al.Wnt signaling:a promising target for osteoarthritis therapy[J].Cell Commun Signal,2019,17(1):97.
[4] ZHENG D,YANG K,CHEN T,et al.Inhibition of Lnc-RNA SNHG14 protects chondrocyte from injury in osteoarthritis via sponging miR-137[J].Autoimmunity,2023,56(1):2270185.
[5] LIU Z H,QI DD,LI X,et al.LncRNA SNHG14 promotes osteogenic differentiation of human bone marrow-derived mesenchymal stem cells via regulating miR-185-5p/WISP2 axis[J].J Biol Regul Homeost Agents,2021,35(2):605-615.
[6] ZHAO S,MI Y,ZHENG B,et al.Highly-metastatic colorectal cancer cell released miR-181a-5p-rich extracellular vesicles promote liver metastasis by activating hepatic stellate cells and remodelling the tumour microenvironment[J].J Extracell Vesicles,2022,11(1):e12186.
[7] ZHAO P,MA G,MA L.miR-181a-5p targets DDX3 X to inhibit the progression of osteoarthritis via NF-ΚB signaling pathway[J].J Orthop Surg Res,2023,18(1):606.
[8] ZHAO X,PENG Y,WANG M,et al.Methylation of PRDX3 expression alleviate ferroptosis and oxidative stress in patients with osteoarthritis cartilage injury[J].Arch Rheumatol,2025,40(2):197-210.
[9] XU J,MA X.Hsa_circ_0032131 knockdown inhibits osteoarthritis progression via the miR-502-5p/PRDX3 axis[J].Aging,2021,13(11):15100-15113.
[10] CHO Y,JEONG S,KIM H,et al.Disease-modifying therapeutic strategies in osteoarthritis:Current status and future directions[J].Exp Mol Med,2021,53(11):1689-1696.
[11] BARTELL L R,FORTIER L A,BONASSAR L J,et al.Mitoprotective therapy prevents rapid,strain-dependent mitochondrial dysfunction after articular cartilage injury[J].J Orthop Res,2020,38(6):1257-1267.
[12] GBO 2021 Osteoarthritis Collaborators.Global,regional,and national burden of osteoarthritis,1990-2020 and projections to 2050:a systematic analysis for the Global Burden of Disease Study 2021[J].Lancet Rheumatol,2023,5(9):e508-e522.
[13] JIANG S,LIU Y,XU B,et al.Noncoding RNAs:New regulatory code in chondrocyteapoptosis and autophagy[J].Wiley Interdiscip Rev RNA,2020,11(4):e1584.
[14] CHEN H,HE C,LIU Y,et al.LncRNA-GAS5 inhibits expression of miR 103 and ameliorates the articular cartilage in adjuvant-induced arthritis in obese mice[J].Dose Res,2020,18(4):1559325820942718.
[15] ZUO Y,XIONG C,GAN X,et al.LncRNA HAGLR silencing inhibits IL-1β-induced chondrocytes inflammatory injury via miR-130a-3p/JAK1 axis[J].J Orthop Surg Res,2023,18(1):203.
[16] TANG J S,YU H X,RUAN R X,et al.LncRNA SNHG14 delivered by bone marrow mesenchymal stem cells-secreted exosomes regulates osteogenesis and adipogenesis in osteoporosis by mediating the miR-27a-3p/LMNB1 axis[J].Kaohsiung J Med Sci,2025,41(5):e70004.
[17] ZHANG J,LEI H,LI X.LncRNA SNHG14 contributes to proinflammatory cytokine production in rheumatoid arthritis via the regulation of the miR-17-5p/MINK1-JNK pathway[J].Environ Toxicol,2021,36(12):2484-2492.
[18] WANG B,LI J,TIAN F.Downregulation of lncRNA SN-HG14 attenuates osteoarthritis by inhibiting FSTL-1 mediated NLRP3 and TLR4/NF-κB pathway through miR-124-3p[J].Life Sci,2021,270:119143.
[19] LIN R X,ZHAN G F,WU J C,et al.LncRNA SNHG14 sponges miR-206 to affect proliferation,apoptosis,and metastasis of hepatocellular carcinoma cells by regulating SOX9[J].Dig Dis Sci,2022,67(3):936-946.
[20] ZHU Y,HUANG C,ZHANG C,et al.LncRNA MIR200 CHG inhibits EMT in gastric cancer by stabilizing miR-200c from target-directed miRNA degradation[J].NatCommun,2023,14(1):8141.
[21] CAZZANELLI P,WUERTZ-KOZAK K.microRNAs in intervertebral disc degeneration,apoptosis,inflammation,and mechanobiology[J].Int J Mol Sci,2020,21(10):3601.
[22] ZHANG Y,LI S,JIN P,et al.Dual functions of microRNA-17 in maintaining cartilage homeostasis and protection against osteoarthritis[J].NatCommun,2022,13(1):2447.
[23] YANG X,CHEN H,ZHENG H,et al.LncRNA SNHG12 promotes osteoarthritis progression through targeted down-regulation of miR-16-5p[J].Clin Lab,2022,68(1).DOI:10.7754/Clin.Lab.2021.210402.
[24] GUO X,ZHANG J,HAN X,et al.LncRNA SNHG1 delayed fracture healingviamodulating miR-181a-5p/PTEN axis[J].J Invest Surg,2022,35(6):1304-1312.
[25] LIANG X,XU W.miR-181a-5p regulates the proliferation and apoptosis of glomerular mesangial cellsby targeting KLF6[J].Exp Ther Med,2020,20(2):1121-1128.
[26] SU W,GUO Y,WANG Q,et al.YAP1 inhibits the senescence of alveolar epithelial cells by targeting Prdx3 to alleviate pulmonary fibrosis[J].Exp Mol Med,2024,56(7):1643-1654.
[27] LOESER R F,CORYELL P R,ARMSTRONG A R,et al.Overexpression of peroxiredoxin 3 in cartilage reduces the severity of age-related osteoarthritis but not surgically induced osteoarthritis in mice[J].ACR Open Rheumatol,2022,4(5):441-446.
[28] 巩朝阳.抑制VCP通过稳定PRDX3表达减缓椎间盘退变的作用机制研究[D].兰州:兰州大学,2024.
GONG Z Y.Inhibition of VCP alleviates intervertebral di-sc degeneration by stabilizing PRDX3 expression:a mechanistic study[D].Lanzhou University,2024.
[29] LI H,CAO Y,CHANG C,et al.Knockdown of circSOD2 ameliorates osteoarthritis progression via the miR-224-5p/PRDX3 axis[J].J Orthop Surg Res,2023,18(1):432.