MiR-223-3p inhibits inflammation and pyroptosis in monosodium urate-induced rats and fibroblast-like synoviocytes by targeting NLRP3
MiR-223-3p inhibits inflammation and pyroptosis in monosodium urate-induced rats and fibroblast-like synoviocytes by targeting NLRP3
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DOI：
10.1111/cei.13587
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发表时间：
2021-03-09
影响因子：
4.6
通讯作者：
Xie, B.
中科院分区：
文献类型：
--
作者：
Tian, J.;Zhou, D.;Xie, B.
在类风湿关节炎中发现miR-223-3p下调。本研究旨在进一步探讨miR-223-3p在痛风关节炎（GA）中的表达水平及其作用。在建立尿尿酸钠（MSU）诱导的GA大鼠和成纤维细胞样滑膜细胞（FLSs）模型后，记录大鼠足部体积和步态评分，并用miR-223-3p模拟物/抑制剂或NLR家族pyrin结构域含3 （NLRP3）过表达质粒转染FLSs。通过生物信息学和双荧光素酶报告基因发现MiR-223-3p靶点。苏木精、伊红染色观察大鼠关节病理损伤。采用酶联免疫吸附法（ELISA）检测大鼠体内白细胞介素(IL)-1 β、肿瘤坏死因子(TNF)- α和关节弹性酶的水平。采用甲基噻唑四氮唑（MTT）和流式细胞术检测FLSs的活力和凋亡情况。采用逆转录-定量聚合酶链式反应（RT-qPCR）、免疫荧光、Western blot或免疫组织化学等方法检测FLSs或大鼠滑膜组织中miR-223-3p、NLRP3、cleaved caspase-1、IL-1 β、凋亡相关斑点样蛋白（AS）和cleaved n端gasdermin D （GSDMD）的表达。MSU增加了大鼠足部体积、步态评分、滑膜组织炎症、IL-1 β、tnf - α和关节弹性蛋白酶水平。MSU降低了FLSs的活力，增加了FLSs的凋亡，上调了NLRP3、ASC、cleaved caspase-1、cleaved n-末端GSDM和IL-1 β的表达，下调了大鼠关节和FLSs滑膜组织中miR-223-3p的表达。MiR-223-3p mimic逆转了MSU降低细胞活力的作用，增加了FLSs的焦亡，而MiR-223-3p抑制剂进一步增强了MSU对FLSs的作用。NLRP3是miR-223-3p的靶标。此外，NLRP3过表达逆转了miR-223-3p对msu诱导的FLSs的影响。MiR-223-3p通过靶向NLRP3抑制msu诱导的大鼠和FLSs的焦亡。
Down-regulated miR-223-3p was found in rheumatoid arthritis. This study aimed to further explore the level and role of miR-223-3p in gout arthritis (GA). After monosodium urate (MSU)-induced GA rat and fibroblast-like synoviocytes (FLSs) models were established, the rat paw volume and gait score were documented and the FLSs were transfected with miR-223-3p mimic/inhibitor or NLR family pyrin domain containing 3 (NLRP3) over-expression plasmids. The MiR-223-3p target was found through bioinformatics and the dual-luciferase reporter. The rat joint pathological damage was observed by hematoxylin and eosin staining. The levels of interleukin (IL)-1 beta, tumor necrosis factor (TNF)-alpha and articular elastase in rats were detected by enzyme-linked immunosorbent assay (ELISA). The viability and pyroptosis of FLSs were detected by methyl thiazolyl tetrazolium (MTT) and flow cytometry. The expressions of miR-223-3p, NLRP3, cleaved caspase-1, IL-1 beta, apoptosis-associated speck-like protein (AS) and cleaved N-terminal gasdermin D (GSDMD) in FLSs or rat synovial tissues were detected by reverse transcription-quantitative polymerase chain reaction (RT-qPCR), immunofluorescence, Western blot or immunohistochemistry analysis. MSU increased the paw volume, gait score, inflammation in synovial tissues and increased the levels of IL-1 beta, TNF-alpha and articular elastase in rats. MSU decreased the viability and increased the pyroptosis of FLSs, up-regulated the expression of NLRP3, ASC, cleaved caspase-1, cleaved N-terminal GSDM, and IL-1 beta, and down-regulated miR-223-3p expression in synovial tissues of rat joints and FLSs. MiR-223-3p mimic reversed the effect of MSU on lowering cell viability, increasing pyroptosis in FLSs, while miR-223-3p inhibitor further enhanced the effect of MSU on FLSs. NLRP3 was a target of miR-223-3p. Also, NLRP3 over-expression reversed the effects of miR-223-3p on MSU-induced FLSs. MiR-223-3p inhibited pyroptosis in MSU-induced rats and FLSs by targeting NLRP3.