miR-185-5p通过靶向抑制IL-1β减轻急性痛风性关节炎的炎症反应
miR-185-5p alleviates the inflammatory response of acute gouty arthritis by inhibiting of IL-1β
侯楠 1马祥会 2周玮 1袁敏 1徐黎明 1孙焕霞 1刘一帆 1刘立宁 1石颜军 1李春先 1付衍法1
作者信息
- 1. 聊城市人民医院风湿免疫科,山东聊城 252000
- 2. 东营市人民医院风湿免疫科,山东东营 257091
- 折叠
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1 β(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.
Abstract
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-1β expression 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 factor α(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-1β expression 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-1β was 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.
关键词
急性痛风性关节炎(AGA)/miR-185-5p/白细胞介素1β(IL-1β)/视觉模拟评分法Key words
acute gouty arthritis(AGA)/miR-185-5p/interleukin-1β/visual analogue scale引用本文复制引用
出版年
2024
NETL
NSTL
万方数据