长链非编码 RNA ANRIL 通过 miR-122-5p/BRCC3 轴促进尿酸肾病中 NLRP3 炎性小体的激活。
LncRNA ANRIL promotes NLRP3 inflammasome activation in uric acid nephropathy through miR-122-5p/BRCC3 axis.
机构信息
Department of Traditional Chinese Medicine, Renmin Hospital of Wuhan University, Wuhan, China.
Department of Traditional Chinese Medicine, Renmin Hospital of Wuhan University, Wuhan, China.
出版信息
Biochimie. 2019 Feb;157:102-110. doi: 10.1016/j.biochi.2018.10.011. Epub 2018 Oct 19.
This study is designed to explore the mechanism by which long non-coding RNA (lncRNA) 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, BRCA1-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.
这项研究旨在探索长链非编码 RNA（lncRNA）反义非编码 RNA 在 INK4 基因座（ANRIL）中在尿酸肾病（UAN）中的致病作用机制。通过 qRT-PCR 测定 UAN 患者和尿酸处理的 HK-2 细胞中 ANRIL、miR-122-5p、BRCA1-BRCA2 含有复合物亚基 3（BRCC3）和 NOD 样受体蛋白 3（NLRP3）的表达。通过 Western blot 检测 BRCC3 和 NLRP3 的蛋白水平。通过 ELISA 定量测定炎症细胞因子水平。使用 CCK-8 测定评估细胞活力。通过 Annexin V-FITC/PI 双标记流式细胞术和 TUNEL 测定检测细胞凋亡。通过荧光素酶报告测定研究 ANRIL、miR-122-5p 和 BRCC3 之间的相互作用。通过实验大鼠评估 ANRIL 在肾损伤中的作用。在 UAN 患者和尿酸处理的管状上皮细胞的血清中，ANRIL 和 BRCC3 表达上调，而 miR-122-5p 下调。荧光素酶报告测定和体外挽救实验证实，ANRIL 通过上调 BRCC3 表达通过海绵 miR-122-5p 促进 NLRP3 炎性小体激活。此外，体内实验验证了敲低 ANRIL 减轻 UAN 大鼠的肾损伤。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。
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