Department of Traditional Chinese Medicine, Renmin Hospital of Wuhan University, Wuhan, China.
Department of Traditional Chinese Medicine, Renmin Hospital of Wuhan University, Wuhan, China.
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。
