摘要
目的 探索白细胞介素1β(IL-1β)在急性痛风性关节炎(AGA)的关节损伤过程中与miR-185-5p的关系。方法 采用实时荧光定量PCR检测89名AGA患者及91名健康志愿者的血清miR-185-5p的水平,采用Pearson相关系数法评估miR-185-5p的表达水平与视觉模拟(VAS)评分以及IL-1β水平之间的相关性;采用受试者工作特征(ROC)曲线评估miR-185-5p对AGA的诊断价值。采用尿酸钠(MSU)诱导THP-1细胞建立体外急性痛风性炎症细胞模型,通过体外转染miR-185-5p模拟物或抑制物增加或降低THP-1细胞miR-185-5p的表达水平后,采用ELISA检测THP-1细胞白细胞介素1β(IL-1β)、 IL-8及肿瘤坏死因子α(TNF-α)的表达水平;荧光素酶报告基因实验确定miR-185-5p与IL-1β的3′-非翻译区(3′-UTR)的相互作用。结果 与健康对照组相比,AGA患者血清miR-185-5p的水平显著降低;血清miR-185-5p的水平与VAS评分和IL-1β的表达水平均呈负相关;ROC曲线下面积为0.905,敏感性为80.17%,特异性为83.52%。下调miR-185-5p显著促进THP-1细胞IL-1β、 IL-8及TNF-α的表达,而过表达miR-185-5p则呈现相反的结果;荧光素酶报告基因实验显示IL-1β是miR-185-5p的靶基因,并且负调控IL-1β的表达。结论 miR-185-5p通过靶向抑制IL-1β减轻AGA的炎症反应。
Objective To investigate the relationship between interleukin-1β(IL-1β)and miR-185-5p in the process of joint injury in acute gouty arthritis(AGA).Methods The serum miR-185-5p levels of 89 AGA patients and 91 healthy volunteers were detected by real-time quantitative PCR.The correlation between miR-185-5p expression level and VAS score or IL-1βexpression level was evaluated by Pearson correlation coefficient method.Receiver operating characteristic(ROC)curve was used to evaluate the diagnostic value of miR-185-5p in AGA.THP-1 cells were induced by sodium urate(MSU)to construct an in vitro acute gouty inflammatory cell model.After the expression level of miR-185-5p in THP-1 cells was upregulated or downregulated by transfection of miR-185-5p mimics or inhibitors in vitro,inflammatory cytokines of THP-1 cells,such as IL-1β,IL-8 and tumor necrosis factorα(TNF-α),were detected by ELISA.The luciferase reporter gene assay was used to determine the interaction between miR-185-5p and the 3'-UTR of IL-1β.Results Compared with the healthy control group,the expression level of serum miR-185-5p in AGA patients was significantly reduced.The level of serum miR-185-5p was negatively correlated with VAS score and IL-1βexpression level.The area under the curve(AUC)was 0.905,the sensitivity was 80.17%and the specificity was 83.52%.Down-regulation of miR-185-5p significantly promoted the expression of IL-1β,IL-8 and tumor necrosis factor(TNF-α),while overexpression of miR-185-5p showed the opposite results.Luciferase reporter gene assay showed that IL-1βwas the target gene of miR-185-5p,and miR-185-5p negatively regulated the expression of IL-1β.Conclusion miR-185-5p alleviates the inflammatory response in AGA by inhibiting IL-1β.
作者
侯楠
马祥会
周玮
袁敏
徐黎明
孙焕霞
刘一帆
刘立宁
石颜军
李春先
付衍法
HOU Nan;MA Xianghui;ZHOU Wei;YUAN Min;XU Liming;SUN Huanxia;LIU Yifan;LIU Lining;SHI Yanjun;LI Chunxian;FU Yanfa(Department of Rheumatology and Immunology,Liaocheng People’s Hospital,Liaocheng 252000;Department of Rheumatology and Immunology,Dongying People’s Hospital,Dongying 257091,China)
出处
《细胞与分子免疫学杂志》
CAS
CSCD
北大核心
2024年第1期51-57,共7页
Chinese Journal of Cellular and Molecular Immunology
基金
山东省中医药科技项目(2021Q010)。
