LncRNA ANRIL promotes NLRP3 inflammasome activation in uric acid nephropathy through miR-122-5p/BRCC3 axis
LncRNA ANRIL promotes NLRP3 inflammasome activation in uric acid nephropathy through miR-122-5p/BRCC3 axis
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DOI：
10.1016/j.biochi.2018.10.011
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发表时间：
2019-02-01
期刊：
影响因子：
3.9
通讯作者：
Dong, Junjun
中科院分区：
文献类型：
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作者：
Hu, Jiacai;Wu, Hao;Dong, Junjun
本研究旨在探讨INK4基因座的长非编码RNA(IncRNA)反义非编码RNA(ANRIL)在尿酸肾病(UAN)发病机制中的作用。采用定量逆转录-聚合酶链式反应(qRT-PCR)检测尿酸处理的HK-2细胞和UAN患者ANRIL、miR-122-5p、BRCA1-BRCA2复合亚单位3(BRCC3)和Nod样受体蛋白3(NLRP3)的表达。免疫印迹法检测BRCC3和NLRP3蛋白水平。用双抗体夹心法测定炎性细胞因子水平。CCK-8比色法检测细胞活力。Annexin V-FITC/PI双标流式细胞术和TUNEL法检测细胞凋亡。用荧光素酶报告基因技术研究了ANRIL、miR-122-5p与BRCC3之间的相互作用。观察ANRIL在实验性大鼠肾损伤中的作用。ANRIL和BRCC3在UAN患者血清和尿酸处理的肾小管上皮细胞中高表达，miR-122-5p表达下调。荧光素酶报告实验和体外挽救实验证实，ANRIL通过海绵miR-122-5p上调BRCC3的表达，促进NLRP3炎症小体的激活。体内实验进一步证实，ANRIL基因敲除可减轻UAN大鼠的肾脏损伤。ANRIL通过miR-122-5p/BRCC3轴在UAN中发挥致病作用，促进NLRP3炎性小体激活。(C)2018年爱思唯尔公司和法国兴业银行(SFBBM)。版权所有。
This study is designed to explore the mechanism by which long non-coding RNA (IncRNA) antisense non-coding RNA in the INK4 locus (ANRIL) plays a pathogenic role in uric acid nephropathy (UAN). The expressions of ANRIL, miR-122-5p, BRCAl-BRCA2-containing complex subunit 3 (BRCC3) and NOD-like receptor protein 3 (NLRP3) were determined in UAN patients and uric acid-treated HK-2 cells by qRT-PCR. Protein levels of BRCC3 and NLRP3 were examined by western blot. The levels of inflammatory cytokines were quantified by ELISA. CCK-8 assay was used to assess cell viability. Apoptosis was detected by Annexin V-FITC/PI double-labeled flow cytometry and TUNEL assay. The interaction between ANRIL, miR-122-5p and BRCC3 were studied using luciferase reporter assay. The role of ANRIL in renal injury was evaluated in experimental rats. ANRIL and BRCC3 were highly expressed while miR-122-5p was down-regulated in serum of UAN patients and uric acid-treated tubular epithelial cells. Luciferase reporter assay and in vitro rescue experiment confirmed that ANRIL promoted NLRP3 inflammasome activation by up-regulating BRCC3 expression via sponging miR-122-5p. Furthermore, in vivo experiment validated that knockdown of ANRIL alleviated renal injury of UAN rats. ANRIL exerted pathogenic effect in UAN to promote NLRP3 inflammasome activation via miR-122-5p/BRCC3 axis. (C) 2018 Elsevier B.V. and Societe Francaise de Biochimie et Biologie Moleculaire (SFBBM). All rights reserved.